Updated on 2021/10/21

写真a

 
SENO Masaharu
 
Organization
Interdisciplinary Science and Engineering in Health Systems Professor
Position
Professor
External link

Degree

  • Ph. D. ( Osaka University )

Research Interests

  • 再生医療

  • Biotechnology

  • Molecular Cell Biology

  • DDS

  • Bioinformatics

  • Tissue regeneration

  • がん幹細胞

  • 生物工学

  • 分子細胞生物学

  • ドラッグデリバリーシステム

  • バイオインフォマティクス

  • Cancer Stem Cells

Research Areas

  • Informatics / Life, health and medical informatics

  • Life Science / Biomedical engineering

  • Life Science / Biomaterials

  • Manufacturing Technology (Mechanical Engineering, Electrical and Electronic Engineering, Chemical Engineering) / Biofunction and bioprocess engineering

  • Life Science / Cell biology

Professional Memberships

  • Japanese Association of Molecular Biology

      More details

  • New York Academy of Science

      More details

  • American Society of Cell Biology

      More details

  • Japanese Cancer Association

      More details

  • 日本分子生物学会

      More details

  • New York Academy of Science

      More details

  • American Society of Cell Biology

      More details

  • 日本再生医療学会

      More details

  • 日本癌学会

      More details

  • 日本バイオインフォマティックス学会

      More details

  • The New York Academy of Sciences ニューヨーク科学アカデミー

      More details

  • American Association of Cancer Research

      More details

  • 米国細胞生物学会(American Association for Cell Biology)

      More details

▼display all

Committee Memberships

  • 日本バイオインフォマティックス学会   中国・四国地域部会長  

    2005   

      More details

    Committee type:Academic society

    日本バイオインフォマティックス学会

    researchmap

 

Papers

  • Microenvironment of mammary fat pads affected the characteristics of the tumors derived from the induced cancer stem cells.

    Abu Quora HA, Zahra MH, El-Ghlban S, Nair N, Afify SM, Hassan G, Nawara HM, Sheta M, Monzur S, Fu X, Osman A, Seno A, Seno M.

    Am J Cancer Res   2021.7

  • Metformin suppresses self-renewal and stemness of cancer stem cell models derived from pluripotent stem cells.

    Zahra MH, Afify SM, Hassan G, Nawara HM, Kumon K, Seno A, Seno M

    Cell Biochem Funct.   2021.7

  • Cripto-1 as a Potential Target of Cancer Stem Cells for Immunotherapy.

    Hiroko Ishii, Said M. Afify, Ghmkin Hassan, David S. Salomon, Masaharu Seno

    Cancers (Basel)   2021.5

  • Paclitaxel-Based Chemotherapy Targeting Cancer Stem Cells from Mono- to Combination Therapy.

    Nawara HM, Afify SM, Hassan G, Zahra MH, Seno A, Seno M.

    Biomedicines   2021.5

  • How can we turn the PI3K/AKT/mTOR pathway down? Insights into inhibition and treatment of cancer.

    Afify SM, Oo AKK, Hassan G, Seno A, Seno M.

    Expert Rev Anticancer Ther.   2021.4

  • Cancer Stem Cell Initiation by Tumor-Derived Extracellular Vesicles.

    Afify SM, Hassan G, Yan T, Seno A, Seno M

    Methods Mol Biol   2021.3

  • Isolation and characterization of cancer stem cells derived from human glioblastoma

    Hiroko Ishii1, Yuki Mimura, Maram H. Zahra, Shota Katayama, Ghmkin Hassan, Said M. Affify, Masaharu Seno

    Am J Cancer Res   2021.2

  • A Novel Artificially Humanized AntiCripto-1 Antibody Suppressing Cancer Cell Growth.

    Ishii, H.; Zahra, M.H.; Takayanagi, A.; Seno, M.

    Int. J. Mol. Sci.   2021.2

  • Cancer Stem Cell Microenvironment Models with Biomaterial Scaffolds In Vitro.

    Ghmkin Hassan, Said M. Afify. Kitano, S.; Seno, A.; Ishii, H.; Shang, Y.; Matsusaki, M.; Seno, M

    Processes 2021   2020.12

  • An assay for cancer stem cell-induced angiogenesis on chick chorioallantoic membrane.

    Nawara HM, Afify SM, Hassan G, Zahra MH, Atallah MN, Seno A, Seno M

    Cell Biol Int   2020.12

  • Tumor-associated macrophages derived from cancer stem cells.

    Osman A, Oze M, Afify SM, Hassan G, El-Ghlban S, Nawara HM, Fu X, Zahra MH, Seno A, Winer I, Salomon DS, Seno M.

    Acta Histochem   2020.9

  • Metastasis Model of Cancer Stem Cell-Derived Tumors.

    Mansour H, Hassan G, Afify SM, Yan T, Seno A, Seno M

    Methods Protoc.   2020.8

     More details

    Language:English  

  • Signaling Inhibitors Accelerate the Conversion of mouse iPS Cells into Cancer Stem Cells in the Tumor Microenvironment Reviewed

    Du, J, Xu, Y, Sasada,, S. Oo, AKK, Hassan, G, Hafizah Mahmud, Apriliana Cahya Khayrani, Md Jahangir Alam, Kazuki Kumon, Ryo Uesaki, Said M. Afify, Hager M. Mansour, Neha Nair, Maram H. Zahra, Akimasa Seno, Nobuhiro Okada, Ling Chen, Ting Yan, Masaharu Seno

    Scientific Reports   2020.6

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Paclitaxel and Sorafenib: The Effective Combination of Suppressing the Self-Renewal of Cancer Stem Cells. Reviewed International journal

    Hend M Nawara, Said M Afify, Ghmkin Hassan, Maram H Zahra, Marwa N Atallah, Hager Mansour, Hagar A Abu Quora, Md Jahangir Alam, Amira Osman, Hiroki Kakuta, Hiroki Hamada, Akimasa Seno, Masaharu Seno

    Cancers   12 ( 6 )   2020.5

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    "Combination therapy", which is a treatment modality combining two or more therapeutic agents, is considered a cornerstone of cancer therapy. The combination of anticancer drugs, of which functions are different from the other, enhances the efficiency compared to the monotherapy because it targets cancer cells in a synergistic or an additive manner. In this study, the combination of paclitaxel and sorafenib in low concentration was evaluated to target cancer stem cells, miPS-BT549cmP and miPS-Huh7cmP cells, developed from mouse induced pluripotent stem cells. The synergistic effect of paclitaxel and sorafenib on cancer stem cells was assessed by the inhibition of proliferation, self-renewal, colony formation, and differentiation. While the IC50 values of paclitaxel and sorafenib were approximately ranging between 250 and 300 nM and between 6.5 and 8 µM, respectively, IC50 of paclitaxel reduced to 20 and 25 nM, which was not toxic in a single dose, in the presence of 1 µM sorafenib, which was not toxic to the cells. Then, the synergistic effect was further assessed for the potential of self-renewal of cancer stem cells by sphere formation ability. As a result, 1 µM of sorafenib significantly enhanced the effect of paclitaxel to suppress the number of spheres. Simultaneously, paclitaxel ranging in 1 to 4 nM significantly suppressed not only the colony formation but also the tube formation of the cancer stem cells in the presence of 1 µM sorafenib. These results suggest the combination therapy of paclitaxel and sorafenib in low doses should be an attractive approach to target cancer stem cells with fewer side effects.

    DOI: 10.3390/cancers12061360

    PubMed

    researchmap

  • Blood and cancer: Blood and Cancer: Cancer Stem Cells as Origin of Hematopoietic Cells in Solid Tumor Microenvironments. Reviewed

    Hassan G, Seno M

    Cells   2020.5

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Cancer stem cell generation by silenced MAPK enhancing PI3K/AKT signaling. Reviewed

    Hassan G, Du J, Afify SM, Seno A, Seno M

    Medical Hypotheses   2020.4

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Revisiting Cancer Stem Cells as the Origin of Cancer-Associated Cells in the Tumor Microenvironment: A Hypothetical View from the Potential of iPSCs. Reviewed

    Osman A, Afify SM, Hassan G, Fu X, Seno A, Seno M

    Cancers (Basel)   2020.4

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • A novel model of liver cancer stem cells developed from induced pluripotent stem cells. Reviewed

    Afify SM, Sanchez Calle A, Hassan G, Kumon K, Nawara HM, Zahra MH, Mansour HM, Khayrani AC, Alam MJ, Du J, Seno A, Iwasaki Y, Seno M

    British Journal of Cancer   2020.4

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Upregulated Ccl20 and Ccr6 in cancer stem cells converted from mouse iPS cells. Reviewed

    Du J, Seno A, Sasada S, Xu Y, Oo AKK, Hassan G, Ueno S, Afify SM, Zahra MH, Okada N, Chen L, Fu X, Tokutaka H, Yan T, SenoM

    Journal of Research in Medical and Dental Science   2020.1

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Hematopoietic Cells Derived from Cancer Stem Cells Generated from Mouse Induced Pluripotent Stem Cells. Reviewed International journal

    Ghmkin Hassan, Said M Afify, Neha Nair, Kazuki Kumon, Amira Osman, Juan Du, Hager Mansour, Hagar A Abu Quora, Hend M Nawara, Ayano Satoh, Maram H Zahra, Nobuhiro Okada, Akimasa Seno, Masaharu Seno

    Cancers   12 ( 1 )   2019.12

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Cancer stem cells (CSCs) represent the subpopulation of cancer cells with the ability to differentiate into other cell phenotypes and initiated tumorigenesis. Previously, we reported generating CSCs from mouse induced pluripotent stem cells (miPSCs). Here, we investigated the ability of the CSCs to differentiate into hematopoietic cells. First, the primary cells were isolated from malignant tumors that were formed by the CSCs. Non-adherent cells (NACs) that arose from adherent cells were collected and their viability, as well as the morphology and expression of hematopoietic cell markers, were analyzed. Moreover, NACs were injected into the tail vein of busulfan conditioned Balb/c nude mice. Finally, CSCs were induced to differentiate to macrophages while using IL3 and SCF. The round nucleated NACs were found to be viable, positive for hematopoietic lineage markers and CD34, and expressed hematopoietic markers, just like homing to the bone marrow. When NACs were injected into mice, Wright-Giemsa staining showed that the number of white blood cells got higher than those in the control mice after four weeks. CSCs also showed the ability to differentiate toward macrophages. CSCs were demonstrated to have the potential to provide progenies with hematopoietic markers, morphology, and homing ability to the bone marrow, which could give new insight into the tumor microenvironment according to the plasticity of CSCs.

    DOI: 10.3390/cancers12010082

    PubMed

    researchmap

  • Cancer stem cell induction from mouse embryonic stem cells. Reviewed International journal

    Akimasa Seno, Chikae Murakami, Bishoy El-Aarag, Yoshiaki Iwasaki, Toshiaki Ohara, Masaharu Seno

    Oncology letters   18 ( 3 )   2756 - 2762   2019.9

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Although cancers are often removed by surgery and treated by chemotherapy and/or radiation therapies, they often reoccur following treatment due to the presence of resistant residual cells such as cancer stem cells (CSCs). CSCs are characterized by their self-renewal, pluripotency, and tumorigenicity properties, and are promising therapeutic targets for the complete therapy of cancers; however, the number of CSCs in cancer tissue is typically too small to investigate fully. We have previously reported that CSCs could be established from induced pluripotent stem cells (iPSCs) using a conditioned medium during cancer cell culture. In the present study, mouse embryonic stem cells (mESCs) were observed to be converted to CSCs (mES-CSCs). This demonstrated that CSC induction does not exclusively occur following gene editing in somatic cells, and that conditioned medium from cancer cells may contain factors that can induce CSCs. Therefore, not only iPSCs but also mESCs, were demonstrated to be able to produce CSCs as one of the potentials of pluripotency of stem cells, suggesting that the conversion to CSCs is not specific to iPSCs. The resultant mES-CSCs would be also useful to generate tissue specific cancers and these naturally occurring cancers can contribute to drug screenings, but also undergo further investigation in order to reveal cancer mechanisms.

    DOI: 10.3892/ol.2019.10614

    PubMed

    researchmap

  • Metastasis of Cancer Stem Cells Developed in the Microenvironment of Hepatocellular Carcinoma. Reviewed International journal

    Said M Afify, Ghmkin Hassan, Amira Osman, Anna Sanchez Calle, Hend M Nawara, Maram Hussein Zahra, Samah El-Ghlban, Hager Mansour, Md Jahangir Alam, Hagar A Abu Quora, Juan Du, Akimasa Seno, Yoshiaki Iwasaki, Masaharu Seno

    Bioengineering (Basel, Switzerland)   6 ( 3 )   2019.8

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Metastasis develops when cancer cells spread from the primary site of a malignant tumor to the surrounding and distant tissues, and it is the most critical problem in cancer treatment. Our group developed cancer stem cells (CSCs) from induced pluripotent stem cells (iPSCs) in the presence of a conditioned medium (CM) of cancer-derived cells. The CSCs were characterized by the formation of malignant tumors in vivo, followed by metastasis. In this study, CSCs converted from mouse iPSCs in the presence of CM from hepatocellular carcinoma (HCC) cell line Huh7 cells. These converted cells (miPS-Huh7cm cells) were established as the metastatic cells. The generated CSCs were injected into the liver or spleen of nude mice. Almost one month after transplantation, the tumors were excised, and the primary cultured cells derived from the malignant tumors and metastatic nodules were evaluated by stemness and metastatic markers to compare their differences. The miPS-Huh7cm cells exhibited metastatic potential, and efficiently formed malignant tumors with lung and/or liver lesions in vivo, whereas the injected miPS formed teratoma. The primary cultured cells derived from the malignant tumors and metastatic nodules sustained the expression of stemness markers, such as Nanog, Klf4 and c-Myc, and acquired cancer stem markers, such as CD90, CD44 and ALDH1. Simultaneously, the expression of metastatic markers, such as Slug, Twist1 and vimentin, in primary cells derived from the malignant tumors, was higher than in metastatic nodules. The CSCs derived from iPSCs, forming malignant tumors and displaying high metastasis, will provide a good animal model to study the mechanisms of metastasis.

    DOI: 10.3390/bioengineering6030073

    PubMed

    researchmap

  • Method to Convert Stem Cells into Cancer Stem Cells. Reviewed

    Afify SM, Chen L, Yan T, Calle AS, Nair N, Murakami C, Zahra MH, Okada N, Iwasaki Y, Seno A, Seno M

    Methods and protocols   2019.8

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Daunorubicin can eliminate iPS-derived cancer stem cells via ICAD/CAD-independent DNA fragmentation. Reviewed

    Akimasa Seno, Akifumi Mizutani, Kazuki Aizawa, Ryoma Onoue, Junko Masuda, Naotaka Ochi, Saki Taniguchi, Tatsuyuki Sota, Yuki Hiramoto, Taisuke Michiue, Neha Nair, Masaharu Seno

    Cancer Drug Resistance   2   335 - 350   2019.6

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.20517/cdr.2019.01

    researchmap

  • Conversion of Stem Cells to Cancer Stem Cells: Undercurrent of Cancer Initiation. Reviewed International journal

    Said M Afify, Masaharu Seno

    Cancers   11 ( 3 )   pii:E345   2019.3

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Cancer stem cells (CSCs) also known as cancer-initiating cells (CIC), are responsible for the sustained and uncontrolled growth of malignant tumors and are proposed to play significant roles in metastasis and recurrence. Several hypotheses have proposed that the events in either stem and/or differentiated cells, such as genomic instability, inflammatory microenvironment, cell fusion, and lateral gene transfer, should be considered as the possible origin of CSCs. However, until now, the exact origin of CSC has been obscure. The development of induced pluripotent stem cells (iPSCs) in 2007, by Yamanaka's group, has been met with much fervency and hailed as a breakthrough discovery by the scientific and research communities, especially in regeneration therapy. The studies on the development of CSC from iPSCs should also open a new page of cancer research, which will help in designing new therapies applicable to CSCs. Currently most reviews have focused on CSCs and CSC niches. However, the insight into the niche before the CSC niche should also be of keen interest. This review introduces the novel concept of cancer initiation introducing the conversion of iPSCs to CSCs and proposes a relationship between the inflammatory microenvironment and cancer initiation as the key concept of the cancer-inducing niche responsible for the development of CSC.

    DOI: 10.3390/cancers11030345

    PubMed

    researchmap

  • Targeting Ovarian Cancer Cells Overexpressing CD44 with Immunoliposomes Encapsulating Glycosylated Paclitaxel. Reviewed International journal

    Apriliana Cahya Khayrani, Hafizah Mahmud, Aung Ko Ko Oo, Maram H Zahra, Miharu Oze, Juan Du, Md Jahangir Alam, Said M Afify, Hagar A Abu Quora, Tsukasa Shigehiro, Anna Sanchez Calle, Nobuhiro Okada, Akimasa Seno, Koki Fujita, Hiroki Hamada, Yuhki Seno, Tadakatsu Mandai, Masaharu Seno

    International journal of molecular sciences   20 ( 5 )   pii:E1042   2019.2

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Paclitaxel (PTX) is one of the front-line drugs approved for the treatment of ovarian cancer. However, the application of PTX is limited due to the significant hydrophobicity and poor pharmacokinetics. We previously reported target-directed liposomes carrying tumor-selective conjugated antibody and encapsulated glycosylated PTX (gPTX-L) which successfully overcome the PTX limitation. The tubulin stabilizing activity of gPTX was equivalent to that of PTX while the cytotoxic activity of gPTX was reduced. In human ovarian cancer cell lines, SK-OV-3 and OVK18, the concentration at which cell growth was inhibited by 50% (IC50) for gPTX range from 15⁻20 nM, which was sensitive enough to address gPTX-L with tumor-selective antibody coupling for ovarian cancer therapy. The cell membrane receptor CD44 is associated with cancer progression and has been recognized as a cancer stem cell marker including ovarian cancer, becoming a suitable candidate to be targeted by gPTX-L therapy. In this study, gPTX-loading liposomes conjugated with anti-CD44 antibody (gPTX-IL) were assessed for the efficacy of targeting CD44-positive ovarian cancer cells. We successfully encapsulated gPTX into liposomes with the loading efficiency (LE) more than 80% in both of gPTX-L and gPTX-IL with a diameter of approximately 100 nm with efficacy of enhanced cytotoxicity in vitro and of convenient treatment in vivo. As the result, gPTX-IL efficiently suppressed tumor growth in vivo. Therefore gPTX-IL could be a promising formulation for effective ovarian cancer therapies.

    DOI: 10.3390/ijms20051042

    PubMed

    researchmap

  • A Novel Combination Cancer Therapy with Iron Chelator Targeting Cancer Stem Cells via Suppressing Stemness. Reviewed International journal

    Yuki Katsura, Toshiaki Ohara, Kazuhiro Noma, Takayuki Ninomiya, Hajime Kashima, Takuya Kato, Hiroaki Sato, Satoshi Komoto, Toru Narusaka, Yasuko Tomono, Boyi Xing, Yuehua Chen, Hiroshi Tazawa, Shunsuke Kagawa, Yasuhiro Shirakawa, Tomonari Kasai, Masaharu Seno, Akihiro Matsukawa, Toshiyoshi Fujiwara

    Cancers   11 ( 2 )   pii:E177   2019.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Excess iron causes cancer and is thought to be related to carcinogenesis and cancer progression including stemness, but the details remain unclear. Here, we hypothesized that stemness in cancer is related to iron metabolism and that regulating iron metabolism in cancer stem cells (CSCs) may be a novel therapy. In this study, we used murine induced pluripotent stem cells that expressed specific stem cell genes such as Nanog, Oct3/4, Sox2, Klf4, and c-Myc, and two human cancer cell lines with similar stem cell gene expression. Deferasirox, an orally available iron chelator, suppressed expression of stemness markers and spherogenesis of cells with high stemness status in vitro. Combination therapy had a marked antitumor effect compared with deferasirox or cisplatin alone. Iron metabolism appears important for maintenance of stemness in CSCs. An iron chelator combined with chemotherapy may be a novel approach via suppressing stemness for CSC targeted therapy.

    DOI: 10.3390/cancers11020177

    PubMed

    researchmap

  • Use of DNA-generated gold nanoparticles to radiosensitize and eradicate radioresistant glioma stem cells. Reviewed International journal

    Tatsuki Kunoh, Tsutomu Shimura, Tomonari Kasai, Syuji Matsumoto, Hafizah Mahmud, Apriliana Cahya Khayrani, Masaharu Seno, Hitoshi Kunoh, Jun Takada

    Nanotechnology   30 ( 5 )   055101 - 055101   2019.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The surface reactivity of gold nanoparticles (AuNPs) is receiving attention as a radiosensitizer of cancer cells for radiation therapy and/or as a drug carrier to target cells. This study demonstrates the potential of DNA-AuNPs (prepared by mixing calf thymus DNA with HAuCl4 solution) as a radiosensitizer of human glioma cells that have cancer stem cell (CSC)-like properties, to reduce their survival. CSC-like U251MG-P1 cells and their parental glioblastoma U251MG cells are treated with a prepared DNA-AuNP colloid. The radiosensitivity of the resultant AuNP-associated cells are significantly enhanced. To reveal the mechanism by which survival is reduced, the generation of reactive oxygen species (ROS), apoptosis induction, or DNA damage in the cells is assayed using the fluorescent dye DCFDA, annexin V-FITC/PI, and foci formation of γ-H2AX, respectively. X-ray irradiation with administration of AuNPs overcomes the radioresistance of U251MG-P1 cells. It does not induce ROS generation or apoptosis in the cells but enhances the number of abnormal nuclei with abundant γ-H2AX foci, which is judged as cell death by mitotic catastrophe. The AuNP association with the cells effectively induces mitotic catastrophe in x-ray-irradiated CSC-like cells, implicating that DNA-AuNPs might be a promising tool to develop an efficient radiosensitizer against CSC.

    DOI: 10.1088/1361-6528/aaedd5

    PubMed

    researchmap

  • Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model. Reviewed International journal

    Md Jahangir Alam, Ryota Takahashi, Said M Afify, Aung Ko Ko Oo, Kazuki Kumon, Hend M Nawara, Aprilliana Cahya Khayrani, Juan Du, Maram H Zahra, Akimasa Seno, David S Salomon, Masaharu Seno

    International journal of molecular sciences   19 ( 11 )   2018.10

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Cripto-1 is a glycophosphatidylinositol (GPI) anchored signaling protein of epidermal growth factor (EGF)-Cripto-1-FRL1-Cryptic (CFC) family and plays a significant role in the early developmental stages and in the different types of cancer cells, epithelial to mesenchymal transition and tumor angiogenesis. Previously, we have developed cancer stem cells (miPS-LLCcm) from mouse iPSCs by culturing them in the presence of conditioned medium of Lewis Lung Carcinoma (LLC) cells for four weeks. Nodal and Cripto-1 were confirmed to be expressed in miPS-LLCcm cells by quantitative reverse transcription PCR (rt-qPCR) implying that Cr-1 was required in maintaining stemness. To investigate the biological effect of adding exogenous soluble CR-1 to the cancer stem cells, we have prepared a C-terminally truncated soluble form of recombinant human CR-1 protein (rhsfCR-1), in which the GPI anchored moiety was removed by substitution of a stop codon through site-directed mutagenesis. rhsfCR-1 effectively suppressed the proliferation and sphere forming ability of miPS-LLCcm cells in a dose-dependent manner in the range of 0 to 5 µg/mL, due to the suppression of Nodal-Cripto-1/ALK4/Smad2 signaling pathway. Frequency of sphere-forming cells was dropped from 1/40 to 1/69 by rhsfCR-1 at 1 µg/mL. Moreover, rhsfCR-1 in the range of 0 to 1 µg/mL also limited the differentiation of miPS-LLCcm cells into vascular endothelial cells probably due to the suppression of self-renewal, which should reduce the number of cells with stemness property. As demonstrated by a soluble form of exogenous Cripto-1 in this study, the efficient blockade would be an attractive way to study Cripto-1 dependent cancer stem cell properties for therapeutic application.

    DOI: 10.3390/ijms19113345

    PubMed

    researchmap

  • Five Fibrosis Biomarkers Together with Serum Ferritin Level to Diagnose Liver Fibrosis and Cirrhosis. Reviewed International journal

    Said M Afify, Ashraf Tabll, Hend M Nawara, Mohamed El Kassas, Ashraf Elfert, Masaharu Seno, Salah El-Kousy

    Clinical laboratory   64 ( 10 )   1685 - 1693   2018.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    BACKGROUND: Liver fibrosis is a dynamic procedure that results from an irregularity between fibrogenesis and fibrolysis. After time this procedure can lead to cirrhosis of the liver. Liver fibrosis and cirrhosis assessment is very important for both therapeutic decisions and prognostic evaluations. In this study, we tried to use serum ferritin (SF) together with five fibrosis tests (Age-Platelet index (API), aspartate aminotransferase to alanine aminotransferase ratio (AAR), AST to platelet ratio index (APRI), Fibrosis 4 score (FIB-4), and fibro-quotient (Fibro-Q)) to assess liver fibrosis and cirrhosis and estimate possible correlation between inflammation and SF. METHODS: This study was carried out on eighty-eight patients infected with HCV and twenty healthy subjects as a control. Complete blood count (CBC), aspartate aminotransferase (AST), alanine aminotransferase (ALT), antiHCV antibody, detection of HCV RNA by real-time PCR, and serum ferritin (SF) were assessed. Then API, ARR, APRI, FIB-4, and Fibro-Q were calculated. Different fibrosis stages (mild fibrosis stage (F1), moderate fibrosis stage (F2), severe fibrosis stage (F3), cirrhotic stage (F4)) were assessed using transient elastography by Fibro Scan®. RESULTS: FIB-4 index was significantly elevated (p < 0.01) with the progression of liver fibrosis at F1, F2, F3, and F4 when compared to healthy control group. The APRI score elevation between F0 and F3 and between F0 and F4 was significant (p < 0.01). SF was elevated in all fibrosis stages and significantly (p < 0.01) at F3 and F4 compared to controls. CONCLUSIONS: APRI coupled with SF should be the best reliable biomarkers for liver cirrhosis. Simultaneously, from our data SF involved in all stages of inflammation. Therefore, down regulation of ferritin in the early stage of fibrosis should be helpful in decreasing the inflammatory effect of ferritin.

    DOI: 10.7754/Clin.Lab.2018.180502

    PubMed

    researchmap

  • HSP-enriched properties of extracellular vesicles involve survival of metastatic oral cancer cells. Reviewed International journal

    Kisho Ono, Takanori Eguchi, Chiharu Sogawa, Stuart K Calderwood, Junya Futagawa, Tomonari Kasai, Masaharu Seno, Kuniaki Okamoto, Akira Sasaki, Ken-Ichi Kozaki

    Journal of cellular biochemistry   119 ( 9 )   7350 - 7362   2018.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Cancer cells often secrete extracellular vesicles (EVs) that carry heat shock proteins (HSPs) with roles in tumor progression. Oral squamous cell carcinoma (OSCC) belongs to head and neck cancers (HNC) whose lymph-node-metastases often lead to poor prognosis. We have examined the EV proteome of OSCC cells and found abundant secretion of HSP90-enriched EVs in lymph-node-metastatic OSCC cells. Double knockdown of HSP90α and HSP90β, using small interfering RNA significantly reduced the survival of the metastatic OSCC cells, although single knockdown of each HSP90 was ineffective. Elevated expression of these HSP90 family members was found to correlate with poor prognosis of HNC cases. Thus, elevated HSP90 levels in secreted vesicles are potential prognostic biomarkers and therapeutic targets in metastatic OSCC.

    DOI: 10.1002/jcb.27039

    PubMed

    researchmap

  • Up-Regulation of PI 3-Kinases and the Activation of PI3K-Akt Signaling Pathway in Cancer Stem-Like Cells Through DNA Hypomethylation Mediated by the Cancer Microenvironment. Reviewed International journal

    Aung Ko Ko Oo, Anna Sanchez Calle, Neha Nair, Hafizah Mahmud, Arun Vaidyanath, Junya Yamauchi, Aprilliana Cahya Khayrani, Juan Du, Md Jahangir Alam, Akimasa Seno, Akifumi Mizutani, Hiroshi Murakami, Yoshiaki Iwasaki, Ling Chen, Tomonari Kasai, Masaharu Seno

    Translational oncology   11 ( 3 )   653 - 663   2018.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Previously, we have succeeded in converting induced pluripotent stem cells (iPSCs) into cancer stem cells (CSCs) by treating the iPSCs with conditioned medium of Lewis lung carcinoma (LLC) cells. The converted CSCs, named miPS-LLCcm cells, exhibited the self-renewal, differentiation potential, and potential to form malignant tumors with metastasis. In this study, we further characterized miPS-LLCcm cells both in vivo and in vitro. The tumors formed by subcutaneous injection showed the structures with pathophysiological features consisting of undifferentiated and malignant phenotypes generally found in adenocarcinoma. Metastasis in the lung was also observed as nodule structures. Excising from the tumors, primary cultured cells from the tumor and the nodule showed self-renewal, differentiation potential as well as tumor forming ability, which are the essential characters of CSCs. We then characterized the epigenetic regulation occurring in the CSCs. By comparing the DNA methylation level of CG rich regions, the differentially methylated regions (DMRs) were evaluated in all stages of CSCs when compared with the parental iPSCs. In DMRs, hypomethylation was found superior to hypermethylation in the miPS-LLCcm cells and its derivatives. The hypo- and hypermethylated genes were used to nominate KEGG pathways related with CSC. As a result, several categories were defined in the KEGG pathways from which most related with cancers, significant and high expression of components was PI3K-AKT signaling pathway. Simultaneously, the AKT activation was also confirmed in the CSCs. The PI3K-Akt signaling pathway should be an important pathway for the CSCs established by the treatment with conditioned medium of LLC cells.

    DOI: 10.1016/j.tranon.2018.03.001

    PubMed

    researchmap

  • Cytokine Expression and Macrophage Localization in Xenograft and Allograft Tumor Models Stimulated with Lipopolysaccharide. Reviewed International journal

    Junko Masuda, Tsukasa Shigehiro, Takuma Matsumoto, Ayano Satoh, Akifumi Mizutani, Chiho Umemura, Shoki Saito, Mayumi Kijihira, Eiji Takayama, Akimasa Seno, Hiroshi Murakami, Masaharu Seno

    International journal of molecular sciences   19 ( 4 )   2018.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    T cell-deficient mice such as nude mice are often used to generate tumor xenograft for the development of anticancer agents. However, the functionality of the other immune cells including macrophages, dendritic cells (DCs), and myeloid-derived suppressor cells (MDSCs) in the xenograft are largely unknown. Macrophages and dendritic cells (DCs) acquire functionally distinct properties in response to various environmental stimuli; the interaction of these cells with MDSCs in tumor microenvironments regulates cancer progression. Nude mice are less likely to reject human cancer cells because of major histocompatibility complex (MHC) mismatches. The tumor microenvironment in a xenograft, comprising human and mouse cells, exhibits more complex bidirectional signaling and function than that of allograft. Here, we evaluated the differences of myeloid cells between them. Plasma interferon-&gamma; and interleukin-18 concentrations in the xenograft tumor model after lipopolysaccharide (LPS) administration were significantly higher than those in the allograft tumor model. MHC class I, II, and CD80 expression levels were increased in CD11b⁺ and MDSC populations after LPS administration in the spleen of a xenograft tumor model but not in that of an allograft tumor model. Additionally, the number of CD80- and mannose receptor C type 1 (MRC1)-expressing cells was decreased upon LPS administration in the tumor of the xenograft tumor. These results suggest that functions of macrophages and DCs are sustained in the xenograft, whereas their functions in response to LPS were suppressed in the allograft. The findings will encourage the consideration of the effects of myeloid cells in the xenograft for drug development.

    DOI: 10.3390/ijms19041261

    PubMed

    researchmap

  • Targeting Glioblastoma Cells Expressing CD44 with Liposomes Encapsulating Doxorubicin and Displaying Chlorotoxin-IgG Fc Fusion Protein. Reviewed International journal

    Hafizah Mahmud, Tomonari Kasai, Apriliana Cahya Khayrani, Mami Asakura, Aung Ko Ko Oo, Juan Du, Arun Vaidyanath, Samah El-Ghlban, Akifumi Mizutani, Akimasa Seno, Hiroshi Murakami, Junko Masuda, Masaharu Seno

    International journal of molecular sciences   19 ( 3 )   2018.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    We recently have established a successful xenograft model of human glioblastoma cells by enriching hyaluronic acid-dependent spheroid-forming populations termed U251MG-P1 cells from U251MG cells. Since U251MG-P1 cells have been confirmed to express CD44 along with principal stemness marker genes, OCT3/4, SOX2, KLF4 and Nanog, this CD44 expressing population appeared to majorly consist of undifferentiated cells. Evaluating the sensitivity to anti-cancer agents, we found U251MG-P1 cells were sensitive to doxorubicin with IC50 at 200 nM. Although doxorubicin has serious side-effects, establishment of an efficient therapy targeting undifferentiated glioblastoma cell population is necessary. We previously designed a chlorotoxin peptide fused to human IgG Fc region without hinge sequence (M-CTX-Fc), which exhibited a stronger growth inhibitory effect on the glioblastoma cell line A172 than an original chlorotoxin peptide. Combining these results together, we designed M-CTX-Fc conjugated liposomes encapsulating doxorubicin and used U251MG-P1 cells as the target model in this study. The liposome modified with M-CTX-Fc was designed with a diameter of approximately 100-150 nm and showed high encapsulation efficiency, adequate loading capacity of anticancer drug, enhanced antitumor effects demonstrating increasing uptake into the cells in vitro; M-CTX-Fc-L-Dox shows great promise in its ability to suppress tumor growth in vivo and it could serve as a template for targeted delivery of other therapeutics.

    DOI: 10.3390/ijms19030659

    PubMed

    researchmap

  • Iron depletion is a novel therapeutic strategy to target cancer stem cells. Reviewed International journal

    Takayuki Ninomiya, Toshiaki Ohara, Kazuhiro Noma, Yuki Katsura, Ryoichi Katsube, Hajime Kashima, Takuya Kato, Yasuko Tomono, Hiroshi Tazawa, Shunsuke Kagawa, Yasuhiro Shirakawa, Fumiaki Kimura, Ling Chen, Tomonari Kasai, Masaharu Seno, Akihiro Matsukawa, Toshiyoshi Fujiwara

    Oncotarget   8 ( 58 )   98405 - 98416   2017.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Adequate iron levels are essential for human health. However, iron overload can act as catalyst for the formation of free radicals, which may cause cancer. Cancer stem cells (CSCs), which maintain the hallmark stem cell characteristics of self-renewal and differentiation capacity, have been proposed as a driving force of tumorigenesis and metastases. In the present study, we investigated the role of iron in the proliferation and stemness of CSCs, using the miPS-LLCcm cell model. Although the anti-cancer agents fluorouracil and cisplatin suppressed the proliferation of miPS-LLCcm cells, these drugs did not alter the expression of stemness markers, including Nanog, SOX2, c-Myc, Oct3/4 and Klf4. In contrast, iron depletion by the iron chelators deferasirox and deferoxamine suppressed the proliferation of miPS-LLCcm cells and the expression of stemness markers. In an allograft model, deferasirox inhibited the growth of miPS-LLCcm implants, which was associated with decreased expression of Nanog and Sox2. Altogether, iron appears to be crucial for the proliferation and maintenance of stemness of CSCs, and iron depletion may be a novel therapeutic strategy to target CSCs.

    DOI: 10.18632/oncotarget.21846

    PubMed

    researchmap

  • Practical Liposomal Formulation for Taxanes with Polyethoxylated Castor Oil and Ethanol with Complete Encapsulation Efficiency and High Loading Efficiency. Reviewed International journal

    Tsukasa Shigehiro, Junko Masuda, Shoki Saito, Apriliana C Khayrani, Kazumasa Jinno, Akimasa Seno, Arun Vaidyanath, Akifumi Mizutani, Tomonari Kasai, Hiroshi Murakami, Ayano Satoh, Tetsuya Ito, Hiroki Hamada, Yuhki Seno, Tadakatsu Mandai, Masaharu Seno

    Nanomaterials (Basel, Switzerland)   7 ( 10 )   1391 - 1398   2017.9

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Taxanes including paclitaxel and docetaxel are effective anticancer agents preferably sufficient for liposomal drug delivery. However, the encapsulation of these drugs with effective amounts into conventional liposomes is difficult due to their high hydrophobicity. Therefore, an effective encapsulation strategy for liposomal taxanes has been eagerly anticipated. In this study, the mixture of polyethoxylated castor oil (Cremophor EL) and ethanol containing phosphate buffered saline termed as CEP was employed as a solvent of the inner hydrophilic core of liposomes where taxanes should be incorporated. Docetaxel-, paclitaxel-, or 7-oxacetylglycosylated paclitaxel-encapsulating liposomes were successfully prepared with almost 100% of encapsulation efficiency and 29.9, 15.4, or 29.1 mol% of loading efficiency, respectively. We then applied the docetaxel-encapsulating liposomes for targeted drug delivery. Docetaxel-encapsulating liposomes were successfully developed HER2-targeted drug delivery by coupling HER2-specific binding peptide on liposome surface. The HER2-targeting liposomes exhibited HER2-specific internalization and enhanced anticancer activity in vitro. Therefore, we propose the sophisticated preparation of liposomal taxanes using CEP as a promising formulation for effective cancer therapies.

    DOI: 10.3390/nano7100290

    PubMed

    researchmap

  • Synthesis and assembly of Hepatitis B virus envelope protein-derived particles in Escherichia coli. Reviewed International journal

    Hao Li, Keisuke Onbe, Qiushi Liu, Masumi Iijima, Kenji Tatematsu, Masaharu Seno, Hiroko Tada, Shun' Ichi Kuroda

    Biochemical and biophysical research communications   490 ( 2 )   155 - 160   2017.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Hepatitis B virus (HBV) envelope particles have been synthesized in eukaryotic cells (e.g., mammalian cells, insect cells, and yeast cells) as an HB vaccine immunogen and drug delivery system (DDS) nanocarrier. Many researchers had made attempts to synthesize the particles in Escherichia coli for minimize the cost and time for producing HBV envelope particles, but the protein was too deleterious to be synthesized in E. coli. In this study, we generated deletion mutants of HBV envelope L protein (389 amino acid residues (aa)) containing three transmembrane domains (TM1, TM2, TM3). The ΔNC mutant spanning from TM2 to N-terminal half of TM3 (from 237 aa to 335 aa) was found as a shortest form showing spontaneous particle formation. After the N-terminal end of ΔNC mutant was optimized by the N-end rule for E. coli expression, the modified ΔNC mutant (mΔNC) was efficiently expressed as particles in E. coli. The molecular mass of mΔNC particle was approx. 670 kDa, and the diameter was 28.5 ± 6.2 nm (mean ± SD, N = 61). The particle could react with anti-HBV envelope S protein antibody, indicating the particles exhibited S antigenic domain outside as well as HBV envelope particles. Taken together, the E. coli-derived mΔNC particles could be used as a substitute of eukaryotic cell-derived HBV envelope particles for versatile applications.

    DOI: 10.1016/j.bbrc.2017.06.015

    PubMed

    researchmap

  • A cancer stem cell model as the point of origin of cancer-associated fibroblasts in tumor microenvironment. Reviewed International journal

    Neha Nair, Anna Sanchez Calle, Maram Hussein Zahra, Marta Prieto-Vila, Aung Ko Ko Oo, Laura Hurley, Arun Vaidyanath, Akimasa Seno, Junko Masuda, Yoshiaki Iwasaki, Hiromi Tanaka, Tomonari Kasai, Masaharu Seno

    Scientific reports   7 ( 1 )   6838 - 6838   2017.7

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    Cancer-associated fibroblasts (CAFs) are one of the most prominent cell types in the stromal compartment of the tumor microenvironment. CAFs support multiple aspects of cancer progression, including tumor initiation, invasion, and metastasis. The heterogeneous nature of the stromal microenvironment is attributed to the multiple sources from which the cells in this compartment originate. The present study provides the first evidence that cancer stem cells (CSCs) are one of the key sources of CAFs in the tumor niche. We generated CSC-like cells by treating mouse induced pluripotent stem cells with conditioned medium from breast cancer cell lines. The resulting cell population expressed both CSC and pluripotency markers, and the sphere-forming CSC-like cells formed subcutaneous tumors in nude mice. Intriguingly, these CSC-like cells always formed heterogeneous populations surrounded by myofibroblast-like cells. Based on this observation, we hypothesized that CSCs could be the source of the CAFs that support tumor maintenance and survival. To address this hypothesis, we induced the differentiation of spheres and purified the myofibroblast-like cells. The resulting cells exhibited a CAF-like phenotype, suggesting that they had differentiated into the subpopulations of cells that support CSC self-renewal. These findings provide novel insights into the dynamic interplay between various microenvironmental factors and CAFs in the CSC niche.

    DOI: 10.1038/s41598-017-07144-5

    PubMed

    researchmap

  • Tumor growth limited to subcutaneous site vs tumor growth in pulmonary site exhibit differential effects on systemic immunities. Reviewed International journal

    Junko Masuda, Eiji Takayama, Warren Strober, Ayano Satoh, Yuji Morimoto, Yasuko Honjo, Tatsuo Ichinohe, Shin-Ichi Tokuno, Toshiaki Ishizuka, Takahiro Nakata, Akifumi Mizutani, Naoki Umemura, Atsushi Kitani, Ivan J Fuss, Tsukasa Shigehiro, Harumi Kawaki, Masako Mizuno-Kamiya, Nobuo Kondoh, Masaharu Seno

    Oncology reports   38 ( 1 )   449 - 455   2017.7

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    To evaluate systemic immunity associated with tumor growth limited to a subcutaneous site versus growth proceeding at multiple tumor sites, we established syngeneic mouse subcutaneous and pulmonary tumor models by local subcutaneous and intravenous injection of colon carcinoma CT26 cells. We found that splenic myeloid-derived suppressor cell (MDSC) levels were significantly increased in the subcutaneous tumor model but not in the pulmonary tumor model. Furthermore, both CD4+ and CD8+ T cells as well as CD4+ Foxp3+ T cells were significantly decreased in the subcutaneous tumor model and were largely unchanged in the pulmonary tumor model. In addition, the subcutaneous model, but not the pulmonary model, displayed a Th1 polarization bias. This bias was characterized by decreased IL-4, IL-9, and IL-10 production, whereas the pulmonary model displayed increased production of IL-10. These results suggest that the mode of tumor development has differential effects on systemic immunity that may, in turn, influence approaches to treatment of cancer patients.

    DOI: 10.3892/or.2017.5646

    PubMed

    researchmap

  • A Novel Model of Meibomian Gland Dysfunction Induced with Complete Freund's Adjuvant in Rabbits. Reviewed International journal

    Hideki Miyake, Tomoko Oda, Osamu Katsuta, Masaharu Seno, Masatsugu Nakamura

    Vision (Basel, Switzerland)   1 ( 1 )   2017.2

     More details

    Language:English  

    A novel meibomian gland dysfunction (MGD) model induced by the injection of complete Freund's adjuvant (CFA) in rabbits was developed to facilitate the understanding of the pathophysiology of MGD with meibomitis. In addition, we sought to evaluate treatment with steroid eye drops in this model. Male Japanese white rabbits were subcutaneously injected with CFA into the upper eyelid margin. The eyelid margins of the rabbits were chronologically observed through slit lamp examination. The development of meibomitis was assessed through histopathology. We evaluated the effects of topically applied tobramycin/dexamethasone (Tob/Dex) eye drops on the plugged orifices and telangiectasia. After the injection of CFA, slit lamp examination revealed markedly plugged orifices, telangiectasia around the orifices and a toothpaste-like meibum, as compared with the normal eyelids. Histopathology revealed granulation tissue with infiltration of inflammatory cells, hyperkeratinization of the ductal epithelium, and cystic dilatation of ducts in the meibomian gland. The orifices were plugged with a proteinaceous substance. Tob/Dex eye drops significantly suppressed the plugging and telangiectasia around the orifices. Through the injection of CFA, we successfully established a novel rabbit MGD that mimics the symptoms observed in humans meibomitis. This model should be useful in the evaluation of the efficacy of drug candidates.

    DOI: 10.3390/vision1010010

    PubMed

    researchmap

  • Immunoliposomes: Recent Progress of Liposomal Drug Delivery System Coupled with Antibodies. in “Liposomes: Historical, Clinical and Molecular Perspectives

    Shigehiro T, Seno, M

    Liposomes Historical,Clinical and Molecular Perspectives   189 - 209   2017

     More details

  • 腫瘍を標的とした抗体提示リポソームの開発と製剤化

    妹尾昌治, 重廣司

    DDS先端技術の製剤への応用開発   249 - 262   2017

     More details

  • Hyaluronic Acid Mediated Enrichment of CD44 Expressing Glioblastoma Stem Cells in U251MG Xenograft Mouse Model.

    Vaidyanath A, Mahmud HB, Khayrani AC, Oo AK, Seno A, Asakura A, Kasai T, Seno M

    Journal of Stem Cell Research and Therapy   2017

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.4172/2157-7633.1000384

    researchmap

  • Transient Tcf3 Gene Repression by TALE-Transcription Factor Targeting

    Junko Masuda, Hiroshi Kawamoto, Warren Strober, Eiji Takayama, Akifumi Mizutani, Hiroshi Murakami, Tomokatsu Ikawa, Atsushi Kitani, Narumi Maeno, Tsukasa Shigehiro, Ayano Satoh, Akimasa Seno, Vaidyanath Arun, Tomonari Kasai, Ivan J. Fuss, Yoshimoto Katsura, Masaharu Seno

    APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY   180 ( 8 )   1559 - 1573   2016.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:HUMANA PRESS INC  

    Transplantation of hematopoietic stem and progenitor cells (HSCs) i.e., self-renewing cells that retain multipotentiality, is now a widely performed therapy for many hematopoietic diseases. However, these cells are present in low number and are subject to replicative senescence after extraction; thus, the acquisition of sufficient numbers of cells for transplantation requires donors able to provide repetitive blood samples and/or methods of expanding cell numbers without disturbing cell multipotentiality. Previous studies have shown that HSCs maintain their multipotentiality and self-renewal activity if TCF3 transcription function is blocked under B cell differentiating conditions. Taking advantage of this finding to devise a new approach to HSC expansion in vitro, we constructed an episomal expression vector that specifically targets and transiently represses the TCF3 gene. This consisted of a vector encoding a transcription activator-like effector (TALE) fused to a Kruppel-associated box (KRAB) repressor. We showed that this TALE-KRAB vector repressed expression of an exogenous reporter gene in HEK293 and COS-7 cell lines and, more importantly, efficiently repressed endogenous TCF3 in a human B lymphoma cell line. These findings suggest that this vector can be used to maintain multipotentiality in HSC being subjected to a long-term expansion regimen prior to transplantation.

    DOI: 10.1007/s12010-016-2187-4

    Web of Science

    researchmap

  • Meibomian gland dysfunction model in hairless mice bred under special diet limiting lipid content

    Hideki Miyake, Tomoko Oda, Osamu Katsuta, Masaharu Seno, Masatsugu Nakamura

    INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE   57 ( 12 )   2016.9

     More details

    Language:English   Publisher:ASSOC RESEARCH VISION OPHTHALMOLOGY INC  

    Web of Science

    researchmap

  • Characterization of gene expression patterns among artificially developed cancer stem cells using spherical self-organizing map

    Akimasa Seno, Tomonari Kasai, Masashi Ikeda, Arun Vaidyanath, Junko Masuda, Akifumi Mizutani, Hiroshi Murakami, Tetsuya Ishikawa, Masaharu Seno

    Cancer Informatics   15   163 - 178   2016.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Libertas Academica Ltd.  

    We performed gene expression microarray analysis coupled with spherical self-organizing map (sSOM) for artificially developed cancer stem cells (CSCs). The CSCs were developed from human induced pluripotent stem cells (hiPSCs) with the conditioned media of cancer cell lines, whereas the CSCs were induced from primary cell culture of human cancer tissues with defined factors (OCT3/4, SOX2, and KLF4). These cells commonly expressed human embryonic stem cell (hESC)/hiPSC-specific genes (POU5F1, SOX2, NANOG, LIN28, and SALL4) at a level equivalent to those of control hiPSC 201B7. The sSOM with unsupervised method demonstrated that the CSCs could be divided into three groups based on their culture conditions and original cancer tissues. Furthermore, with supervised method, sSOM nominated TMED9, RNASE1, NGFR, ST3GAL1, TNS4, BTG2, SLC16A3, CD177, CES1, GDF15, STMN2, FAM20A, NPPB, CD99, MYL7, PRSS23, AHNAK, and LOC152573 genes commonly upregulating among the CSCs compared to hiPSC, suggesting the gene signature of the CSCs.

    DOI: 10.4137/CIN.S39839

    Scopus

    researchmap

  • Cancer stem cells as the novel origin of cancer-associated fibroblast-like cells

    Neha Nair, Arun Vaidyanath, Kenta Hoshikawa, Anna Sanchez Calle, Tomonari Kasai, Masaharu Seno

    CANCER RESEARCH   76   2016.7

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2016-LB-278

    Web of Science

    researchmap

  • The significance of c-Kit protooncogene in iCSC-derived PDAC model

    Anna Sanchez Calle, Kenta Hoshikawa, Neha Nair, Marta Prieto-Vila, Arun Vaidyanath, Tomonari Kasai, Masaharu Seno

    CANCER RESEARCH   76   2016.7

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2016-1725

    Web of Science

    researchmap

  • Meibomian Gland Dysfunction Model in Hairless Mice Fed a Special Diet With Limited Lipid Content

    Hideki Miyake, Tomoko Oda, Osamu Katsuta, Masaharu Seno, Masatsugu Nakamura

    INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE   57 ( 7 )   3268 - 3275   2016.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ASSOC RESEARCH VISION OPHTHALMOLOGY INC  

    PURPOSE. A novel meibomian gland dysfunction (MGD) model was developed to facilitate understanding of the pathophysiology of MGD and to evaluate treatment with azithromycin ophthalmic solution (azithromycin). MGD was induced in HR-1 hairless mice by feeding them a special diet with limited lipid content (HR-AD).
    MEHODS. Male HR-1 hairless mice were fed an HR-AD diet for 16 weeks. Development of MGD was assessed by histopathology at 4-week intervals. The lid margin was observed by slit-lamp examination. After cessation of the HR-AD diet, the mice were fed a normal diet to restore normal eye conditions. Expression of cytokeratin 6 was determined by immunostaining. We evaluated the effects of topically applied azithromycin on the plugged orifice in this model.
    RESULTS. After mice were fed the HR-AD diet, histopathology analysis showed hyperkeratinization of the ductal epithelium in the meibomian gland. Ductal hyperkeratinization resulted in the loss of acini, followed by atrophy of the gland. Slit-lamp examination revealed a markedly plugged orifice, telangiectasia, and a toothpaste-like meibum compared with that of a normal eyelid. Cessation of feeding with HR-AD ameliorated both the MGD signs and the expression of cytokeratin 6, restoring the tissue to a histologically normal state. Azithromycin treatment significantly decreased the number of plugged orifices and ameliorated atrophy, as revealed by histopathologic analysis.
    CONCLUSIONS. We developed a novel model that mimics human MGD signs in HR-1 hairless mice fed an HR-AD diet. Azithromycin treatment led to therapeutic improvement in this model. This MGD model could be useful for the evaluation of drug candidates for MGD.

    DOI: 10.1167/iovs.16-19227

    Web of Science

    researchmap

  • Synthesis and in vitro cancer cell growth inhibition evaluation of 11-amino-modified 5-Me-indolo[2,3-b]quinolines and their COMPARE analyses

    Masashi Okada, Zhen-Wu Mei, Md. Imran Hossain, Li Wang, Taihei Tominaga, Takeshi Takebayashi, Masaharu Murakami, Mizuki Yasuda, Tsukasa Shigehiro, Tomonari Kasai, Akifumi Mizutani, Hiroshi Murakami, Ibrahim El Tantawy El Sayed, Shingo Dan, Takao Yamori, Masaharu Seno, Tsutomu Inokuchi

    MEDICINAL CHEMISTRY RESEARCH   25 ( 5 )   879 - 892   2016.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER BIRKHAUSER  

    A plant-derived neocryptolepine core, the 5-Me-indolo[2,3-b]quinoline skeleton, was emblazoned with substituents at C11 and C2 and then tested against various cancer cell lines to find potent anticancer agents. In the in vitro antiproliferative activity assay against the breast cancer MDA-MB-453 cell line, the attachment of alkylamino substituents at C11 of the 5-Me-indolo[2,3-b]quinoline induced improved activities. Specifically, 11-(3-aminopropylamino) and 11-(4-aminobutylamino) derivatives indicated the highest activity and selectivity against MDA-MB-453 (IC50 = 0.3-0.5 mu M) and also exhibited a higher cytotoxicity against the colon adenocarcinoma (WiDr) and ovarian cancer (SKOv3) cell lines. A synergistic effect by attachment of substituents at C2 was favorably observed with an electron-donating group, such as CH3O, and unfavorably observed with an electron-withdrawing one, such as F and CF3. Further modification of the terminal free amino group of the lariat attachment at C11 into the corresponding acylamides and 2,3-dihydrobenzo[e][1,3]thiazin-4-ones was not effective for the antiproliferative activity. The computer-assisted database analysis, COMPARE, suggested that 14e and 13b have a mode of action similar to actinomycin D and 13c has a mode of actions similar to vindesine sulfate or aclarubicin hydrochloride. However, the new compounds may have other unique mode of actions since the correlation coefficients (r) were in relatively low levels.

    DOI: 10.1007/s00044-016-1508-z

    Web of Science

    researchmap

  • Evaluation of glycosylated docetaxel-encapsulated liposomes prepared by remote loading under solubility gradient

    Tsukasa Shigehiro, Wenjia Zhai, Arun Vaidyanath, Junko Masuda, Akifumi Mizutani, Tomonari Kasai, Hiroshi Murakami, Hiroki Hamada, David S. Salomon, Katsuhiko Mikuni, Yuhki Seno, Tadakatsu Mandai, Masaharu Seno

    JOURNAL OF MICROENCAPSULATION   33 ( 2 )   172 - 182   2016.2

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    Docetaxel comprises one of the most effective anti-cancer drugs despite of serious side effects. Liposomes encapsulation is practically feasible to deliver the drug. However, due to the significant hydrophobicity, docetaxel will be integrated into the lipid bilayer resulting in poor encapsulation capacity. Here, we evaluated a remote loading strategy using a solubility gradient made between the two solvents for 7-glucosyloxyacetyldocetaxel, which has enhanced water solubility of docetaxel with a coupled glucose moiety. Therefore, 7-glucosyloxyacetyldocetaxel was more effectively encapsulated into liposomes with 71.0% of encapsulation efficiency than docetaxel. While 7-glucosyloxyacetyldocetaxel exhibited 90.9% of tubulin stabilisation activity of docetaxel, 7-glucosyloxyacetyldocetaxel encapsulated in liposomes significantly inhibited the growth of tumour in vivo with side effects less than unencapsulated drug. Collectively, the encapsulation of 7-glucosyloxyacetyldocetaxel into liposomes by remote loading under the solubility gradient is considered to be a promising application to prepare practical drug delivery system.

    DOI: 10.3109/02652048.2016.1144815

    Web of Science

    researchmap

  • Release of siRNA from Liposomes Induced by Curcumin

    Kazuyo Fujita, Yoshie Hiramatsu, Hideki Minematsu, Masaharu Somiya, Shun'ichi Kuroda, Masaharu Seno, Shuji Hinuma

    Journal of Nanotechnology   2016   2016

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Hindawi Limited  

    Liposomes are a potential carrier of small interfering RNA (siRNA) for drug delivery systems (DDS). In this study, we searched for a molecule capable of controlling the release of siRNA from a certain type of liposomes and found that curcumin could induce the release of siRNA from the liposomes encapsulating siRNA within 30 min. However, the release of siRNA from the liposomes by curcumin showed a unique dose-response (i.e., bell-shaped curve) with a maximal induction at around 60 μg/ml of curcumin. Liposomal lipid compositions and temperatures influenced the efficiency in the release of siRNA induced by curcumin. About 10% of curcumin at a 60 μg/ml dose was incorporated into the liposomes within 30 min under our experimental conditions. Our results suggest a possibility that curcumin is useful in controlling the permeability of liposomes carrying large molecules like siRNA.

    DOI: 10.1155/2016/7051523

    Scopus

    researchmap

  • A new PDAC mouse model originated from iPSCs-converted pancreatic cancer stem cells (CSCcm)

    Anna Sanchez Calle, Neha Nair, Aung KoKo Oo, Marta Prieto-Vila, Megumi Koga, Apriliana Cahya Khayrani, Maram Hussein, Laura Hurley, Arun Vaidyanath, Akimasa Seno, Yoshiaki Iwasaki, Malu Calle, Tomonari Kasai, Masaharu Seno

    AMERICAN JOURNAL OF CANCER RESEARCH   6 ( 12 )   2799 - 2815   2016

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:E-CENTURY PUBLISHING CORP  

    Pancreatic ductal adenocarcinoma (PDAC) is the most representative form of pancreatic cancers. PDAC solid tumours are constituted of heterogeneous populations of cells including cancer stem cells (CSCs), differentiated cancer cells, desmoplastic stroma and immune cells. The identification and consequent isolation of pancreatic CSCs facilitated the generation of genetically engineered murine models. Nonetheless, the current models may not be representative for the spontaneous tumour occurrence. In the present study, we show the generation of a novel pancreatic iPSC-converted cancer stem cell lines (CSCcm) as a cutting-edge model for the study of PDAC. The CSCcm lines were achieved only by the influence of pancreatic cancer cell lines conditioned medium and were not subjected to any genetic manipulation. The xenografts tumours from CSCcm lines displayed histopathological features of ADM, PanIN and PDAC lesions. Further molecular characterization from RNA-sequencing analysis highlighted primary culture cell lines (1st CSCcm) as potential candidates to represent the pancreatic CSCs and indicated the establishment of the pancreatic cancer molecular pattern in their subsequent progenies 2nd CSCcm and 3rd CSCcm. In addition, preliminary RNA-seq SNPs analysis showed that the distinct CSCcm lines did not harbour single point mutations for the oncogene Kras codon 12 or 13. Therefore, PDAC-CSCcm model may provide new insights about the actual occurrence of the pancreatic cancer leading to develop different approaches to target CSCs and abrogate the progression of this fatidic disease.

    Web of Science

    researchmap

  • iPSC-derived cancer stem cells provide a model of tumor vasculature

    Marta Prieto-Vila, Ting Yan, Anna Sanchez Calle, Neha Nair, Laura Hurley, Tomonari Kasai, Hiroki Kakuta, Junko Masuda, Hiroshi Murakami, Akifumi Mizutani, Masaharu Seno

    AMERICAN JOURNAL OF CANCER RESEARCH   6 ( 9 )   1906 - 1921   2016

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:E-CENTURY PUBLISHING CORP  

    To grow beyond a size of approximately 1-2 mm(3), tumor cells activate many processes to develop blood vasculature. Growing evidences indicate that the formation of the tumor vascular network is very complex, and is not restricted to angiogenesis. Cancer cell-derived tumor vasculatures have been recently described. Among them, endothelial differentiation of tumor cells have been directly related to cancer stem cells, which are cells within a tumor that possess the capacity to self-renew, and to exhibit multipotential heterogeneous lineages of cancer cells. Vasculogenic mimicry has been described to be formed by cancer cells expressing stemness markers. Thus, cancer stem cells have been proposed to contribute to vasculogenic mimicry, though its relation is yet to be clarified. Here, we analyzed the tumor vasculature by using a model of mouse cancer stem cells, miPS-LLCcm cells, which we have previously established from mouse induced pluripotent stem cells and we introduced the DsRed gene in miPS-LLCcm to trace them in vivo. Various features of vasculature were evaluated in ovo, in vitro, and in vivo. The tumors formed in allograft nude mice exhibited angiogenesis in chick chorioallantoic membrane assay. In those tumors, along with penetrated host endothelial vessels, we detected endothelial differentiation from cancer stem cells and formation of vasculogenic mimicry. The angiogenic factors such as VEGF-A and FGF2 were expressed predominantly in the cancer stem cells subpopulation of miPS-LLCcm cells. Our results suggested that cancer stem cells play key roles in not only the recruitment of host endothelial vessels into tumor, but also in maturation of endothelial linage of cancer stem cell's progenies. Furthermore, the undifferentiated subpopulation of the miPS-LLCcm participates directly in the vasculogenic mimicry formation. Collectively, we show that miPS-LLCcm cells have advantages to further study tumor vasculature and to develop novel targeting strategies in the future.

    Web of Science

    researchmap

  • A Unique Procedure to Identify Cell Surface Markers Through a Spherical Self-Organizing Map Applied to DNA Microarray Analysis.

    Sugii Y, Kasai T, Ikeda M, Vaidyanath A, Kumon K, Mizutani A, Seno A, Tokutaka H, Kudoh T, Seno M

    Biomark Cancer   8   17 - 23   2016

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.4137/BIC.S33542

    researchmap

  • Synthesis, Biological Evaluation, Docking and QSAR Studies of Some Novel Naphthalimide Dithiocarbamate Analogs as Antitumor and Anti-Inflammatory Agents.

    Zahra MH, Osman AMA, Agwa H, Nair N, Calle AS, Hurley L, Farag D, Kasai T, Seno M, Zahran M

    Medicinal Chemistry (Los Angeles)   6 ( 12 )   694 - 703   2016

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • 第3節 iPS細胞を用いたがん幹細胞モデル作成のポイントとその評価, 「次世代のがん治療薬・診断のための研究開発~免疫療法・遺伝子治療・がん幹細胞~、第13章 がん幹細胞を標的とした治療薬研究」

    技術情報協会   348 - 353   2016

     More details

  • 球面自己組織化マップを用いたキナーゼパネルアセイデータのクラスタリング

    工藤孝幸, 妹尾昌治

    CCISJ Bulletin   34 ( 1 )   2 - 5   2016

  • がんモデル動物を用いた悪性度の違いによる全身免疫能変化の解析

    増田 潤子, 高山 英次, 佐藤 あやの, 守本 祐司, 本庶 仁子, 石塚 俊晶, 徳野 慎一, 青笹 季文, 光吉 俊二, 重廣 司, 前野 成実, 村上 宏, 笠井 智成, 水谷 昭文, Vaidyanath Arun, 妹尾 彬正, 川木 晴美, 神谷 真子, 野, 近藤 信夫, 一瀬 雅夫, 一戸 辰夫, 妹尾 昌治

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集   88回・38回   [1P1079] - [1P1079]   2015.12

     More details

    Language:Japanese   Publisher:(公社)日本生化学会  

    researchmap

  • 鉄コントロールによる新規がん幹細胞治療 Reviewed

    二宮 卓之, 大原 利章, 勝部 亮一, 賀島 肇, 加藤 卓也, 野間 和広, 田澤 大, 香川 俊輔, 水谷 昭文, 笠井 智成, 妹尾 昌治, 藤原 俊義

    日本癌学会総会記事   74回   IS4 - 6   2015.10

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Dextran sodium sulfate-induced colitis in dendritic cell deletion mice Reviewed

    Junko Masuda, Atsuhi Kitani, Ivan Fuss, Masaharu Seno, Warren Strober

    CANCER RESEARCH   75   2015.8

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2015-4296

    Web of Science

    researchmap

  • Derivation of a model of cancer stem cell from human induced pluripotent stem cells

    Tomonari Kasai, Kenta Hoshikawa, Shuto Takejiri, Masashi Ikeda, Kazuki Kumon, Anna Sanchez Calle, Arun Vaidyanath, Akifumi Mizutani, Chen Ling, Masaharu Seno

    CANCER RESEARCH   75   2015.8

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2015-LB-144

    Web of Science

    researchmap

  • Iron control is a novel therapeutic target of cancer stem cells

    Takayuki Ninomiya, Toshiaki Ohara, Hajime Kashima, Ryoichi Katsube, Kazuhiro Noma, Yasuko Tomono, Akifumi Mizutani, Tomonari Kasai, Masaharu Seno, Shinji Kuroda, Hiroyuki Kishimoto, Hiroshi Tazawa, Yasuhiro Shirakawa, Shunsuke Kagawa, Toshiyoshi Fujiwara

    CANCER RESEARCH   75   2015.8

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2015-4243

    Web of Science

    researchmap

  • Eosinophil Cationic Protein Shows Survival Effect on H9c2 Cardiac Myoblast Cells with Enhanced Phosphorylation of ERK and Akt/GSK-3 beta under Oxidative Stress

    Hiroko Ishii, Shigeshi Kamikawa, Satoshi Hirohata, Akifumi Mizutani, Koji Abe, Masaharu Seno, Toshitaka Ohashi, Yoshifumi Ninomiya

    ACTA MEDICA OKAYAMA   69 ( 3 )   145 - 153   2015.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    Eosinophil cationic protein (ECP) is well known as a cationic protein contained in the basic granules of activated eosinophils. Recent studies have reported that ECP exhibits novel activities on various types of cells, including rat neonatal cardiomyocytes. Here we evaluated the effects of ECP on rat cardiac myoblast H9c2 cells. Our results showed that ECP enhanced the survival of the cells, in part by promoting the ERK and Akt/GSK-3 beta signaling pathways. ECP attenuated the cytotoxic effects of H2O2 on H9c2 cells as well as the production of reactive oxygen species, the number of apoptotic cells and caspase 3/7 activity in the cells. In conclusion, ECP activated the ERR and Akt/GSK-3 beta pathways, resulting in anti-oxidative effects on H9c2 cells that attenuated apoptosis.

    DOI: 10.18926/AMO/53521

    Web of Science

    researchmap

  • Insight into Cancer Stem Cell Niche; Lessons from Cancer Stem Cell Models Generated In Vitro.

    Biology in Stem Cell Niche, Part of the series Stem Cell Biology and Regenerative Medicine   211 - 226   2015

     More details

  • Spherical Self-Organizing Map Detects MYBL 1 As Candidate Gene for Triple-Negative Breast Cancer.

    Ikeda M, Kumon K, Omoto K, Sugii Y, Mizutani A, Vaidyanath A, Kudoh T, Kasai T, Masuda S, Seno M

    Neurosci Biomed Eng   3 ( 2 )   94 - 101   2015

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Bacteria: Prospective Savior in Battle against Cancer

    Neha Nair, Tomonari Kasai, Masaharu Seno

    ANTICANCER RESEARCH   34 ( 11 )   6289 - 6296   2014.11

     More details

    Language:English   Publisher:INT INST ANTICANCER RESEARCH  

    Conventional anticancer therapies such as chemotherapy are losing their sheen in the battle against cancer. Therefore, strategies for treatment of cancer need to be constantly modified to fulfill the growing demands of alternative therapies. Several viral and non-viral vectors have been exploited for anticancer gene therapy. But over the years bacteria have been proven to be an important candidate for successful evasion of cancer. They serve as invaluable source of tumor-specific anticancer genes, toxins, polysaccharides for synthesis of nanodrugs and gene-delivery vectors. The current review assesses the role of important bacterial groups in different spheres of anti-cancer research.

    Web of Science

    researchmap

  • Cancer stem cells maintain a hierarchy of differentiation by creating their niche Reviewed

    Akifumi Mizutani, Shuichi Matsuda, Ting Yan, Marta Prieto-Vila, Ling Chen, Ayano Satoh, Tomonari Kasai, Junko Masuda, Takyuki Kudoh, Hiroshi Murakami, Li Fu, David S. Salomon, Masaharu Seno

    CANCER RESEARCH   74 ( 19 )   2014.10

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2014-3026

    Web of Science

    researchmap

  • Characterization of cancer stem-like cells derived from mouse induced pluripotent stem cells transformed by tumor-derived exosomes/microvesicles Reviewed

    Yan Ting, Junko Masuda, Akifumi Mizutani, Ling Chen, Tsukasa Shigehiro, Shuichi Matsuda, Tomonari Kasai, Takayuki Kudoh, Hiroshi Murakami, Mary J. C. Hendrix, Luigi Strizzi, David S. Salomon, Li Fu, Masaharu Seno

    CANCER RESEARCH   74 ( 19 )   2014.10

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2014-3016

    Web of Science

    researchmap

  • Anti-cancer activity of immunoliposomes encapsulated effective amount of glycosylated paclitaxel with novel loading strategy

    Tsukasa Shigehiro, Tomonari Kasai, Akifumi Mizutani, Hiroshi Murakami, Katsuhiko Mikuni, Tadakatsu Mandai, Hiroki Hamada, Masaharu Seno

    CANCER RESEARCH   74 ( 19 )   2014.10

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2014-4461

    Web of Science

    researchmap

  • Efficient Drug Delivery of Paclitaxel Glycoside: A Novel Solubility Gradient Encapsulation into Liposomes Coupled with Immunoliposomes Preparation

    Tsukasa Shigehiro, Tomonari Kasai, Masaharu Murakami, Sreeja C. Sekhar, Yuki Tominaga, Masashi Okada, Takayuki Kudoh, Akifumi Mizutani, Hiroshi Murakami, David S. Salomon, Katsuhiko Mikuni, Tadakatsu Mandai, Hiroki Hamada, Masaharu Seno

    PLOS ONE   9 ( 9 )   e107976   2014.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PUBLIC LIBRARY SCIENCE  

    Although the encapsulation of paclitaxel into liposomes has been extensively studied, its significant hydrophobic and uncharged character has generated substantial difficulties concerning its efficient encapsulation into the inner water core of liposomes. We found that a more hydrophilic paclitaxel molecule, 7-glucosyloxyacetylpaclitaxel, retained tubulin polymerization stabilization activity. The hydrophilic nature of 7-glucosyloxyacetylpaclitaxel allowed its efficient encapsulation into the inner water core of liposomes, which was successfully accomplished using a remote loading method with a solubility gradient between 40% ethylene glycol and Cremophor EL/ethanol in PBS. Trastuzumab was then conjugated onto the surface of liposomes as immunoliposomes to selectively target human epidermal growth factor receptor-2 (HER2)-overexpressing cancer cells. In vitro cytotoxicity assays revealed that the immunoliposomes enhanced the toxicity of 7-glucosyloxyacetylpaclitaxel in HER2-overexpressing cancer cells and showed more rapid suppression of cell growth. The immunoliposomes strongly inhibited the tumor growth of HT-29 cells xenografted in nude mice. Notably, mice survived when treated with the immunoliposomes formulation, even when administered at a lethal dose of 7glucosyloxyacetylpaclitaxel in vivo. This data successfully demonstrates immunoliposomes as a promising candidate for the efficient delivery of paclitaxel glycoside.

    DOI: 10.1371/journal.pone.0107976

    Web of Science

    researchmap

  • 鉄コントロールによるがん幹細胞の新規治療法(Iron control is a potent novel therapeutic for cancer stem cells) Reviewed

    二宮 卓之, 大原 利章, 浦野 真一, 勝部 亮一, 野間 和広, 田澤 大, 香川 俊輔, 水谷 昭文, 笠井 智成, 妹尾 昌治, 藤原 俊義

    日本癌学会総会記事   73回   P - 1187   2014.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • In vitro anti-proliferative and anti-angiogenic activities of thalidomide dithiocarbamate analogs

    Bishoy Y. A. El-Aarag, Tomonari Kasai, Magdy A. H. Zahran, Nadia I. Zakhary, Tsukasa Shigehiro, Sreeja C. Sekhar, Hussein S. Agwa, Akifumi Mizutani, Hiroshi Murakami, Hiroki Kakuta, Masaharu Seno

    INTERNATIONAL IMMUNOPHARMACOLOGY   21 ( 2 )   283 - 292   2014.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Inhibition of angiogenesis is currently perceived as a promising strategy in the treatment of cancer. The anti-angiogenicity of thalidomide has inspired a second wave of research on this teratogenic drug. The present study aimed to investigate the anti-proliferative and anti-angiogenic activities of two thalidomide dithiocarbamate analogs by studying their anti-proliferative effects on human umbilical vein endothelial cells (HUVECs) and MDA-MB-231 human breast cancer cell lines. Their action on the expression levels of IL-6, IL-8, TNF-alpha, VEGF(165), and MMP-2 was also assessed. Furthermore, their effect on angiogenesis was evaluated through wound healing, migration, tube formation, and nitric oxide (NO) assays. Results illustrated that the proliferation of HUVECs and MDA-MB-231 cells was not significantly affected by thalidomide at 625-100 mu M. Thalidomide failed to block angiogenesis at similar concentrations. By contrast, thalidomide dithiocarbamate analogs exhibited significant anti-proliferative action on HUVECs and MDA-MB-231 cells without causing cytotoxicity and also showed powerful anti-angiogenicity in wound healing, migration, tube formation, and NO assays. Thalidomide analogs 1 and 2 demonstrated more potent activity to suppress expression levels of IL-6, IL-8, TNF-alpha, VEGF(165), and MMP-2 than thalidomide. Analog 1 consistently, showed the highest potency and efficacy in all the assays. Taken together, our results support further development and evaluation of novel thalidomide analogs as antitumor and anti-angiogenic agents. (C) 2014 The Authors. Published by Elsevier B.V.

    DOI: 10.1016/j.intimp2014.05.007

    Web of Science

    researchmap

  • Cancer stem cells maintain a hierarchy of differentiation by creating their niche. Reviewed

    Int J Cancer   135 ( 1 )   27 - 36   2014.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • A Cancer Stem Cell Model: An Insight into the Conversion of Induced Pluripotent Stem Cells to Cancer Stem-Like Cells

    Akifumi Mizutani, Ling Chen, Tomonari Kasai, Takayuki Kudoh, Hiroshi Murakami, Li Fu, Masaharu Seno

    Cancer Stem Cells   79 - 87   2014.4

     More details

    Language:English   Publishing type:Part of collection (book)   Publisher:Wiley Blackwell  

    Appropriate model cell lines recapitulating cancer stem cell (CSC) properties would accelerate not only investigation of cancer stem cells but also development of new clinical cancer therapy by establishing a screening system for anti-cancer stem cell agents. In this chapter, the authors introduce their recent work and others to generate cells with cancer stem cell properties in vitro. They also discuss the concept of cancer stem cells with results obtained from originally established cancer stem-like cells. Finally, they describe future applications of the model to basic and clinical studies. In the field of regeneration therapy, the pluripotent stem cells such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are promising sources of differentiated cells for transplantation. Interestingly, these cancer stem-like cells were not required to be introduced with the genes for cell surface markers that are commonly used to characterize and isolate cancer stem cells.

    DOI: 10.1002/9781118356203.ch6

    Scopus

    researchmap

  • The new proposal of the calculation for the significance degree by once SOM learning - Using iris, gene, and Tof-SIMS data Reviewed

    M. Oyabu, H. Tokutaka, M. Ohkita, M. Seno, M. Ohki

    2014 Joint 7th International Conference on Soft Computing and Intelligent Systems, SCIS 2014 and 15th International Symposium on Advanced Intelligent Systems, ISIS 2014   1114 - 1119   2014.2

     More details

    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:Institute of Electrical and Electronics Engineers Inc.  

    The significance degree of each component was calculated only by once SOM learning where the Spherical Self-Organizing-Map (SSOM) was used for the demonstration. The method can also be used by the usual planar SOM. In the method, kinds of specimens of the data are inserted in each column as each specimen. The method is first demonstrated using the iris data.

    DOI: 10.1109/SCIS-ISIS.2014.7044648

    Scopus

    researchmap

  • Development of In-111-Labeled Liposomes for Vulnerable Atherosclerotic Plaque Imaging

    Mikako Ogawa, Izumi O. Umeda, Mutsumi Kosugi, Ayumi Kawai, Yuka Hamaya, Misato Takashima, Hongxia Yin, Takayuki Kudoh, Masaharu Seno, Yasuhiro Magata

    JOURNAL OF NUCLEAR MEDICINE   55 ( 1 )   115 - 120   2014.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SOC NUCLEAR MEDICINE INC  

    Macrophage infiltration is a common characteristic feature of atherosclerotic-vulnerable plaques. Macrophages recognize phosphatidylserine (PS) exposed on the surface of apoptotic cells, which triggers the engulfment of the apoptotic cells by macrophages through phagocytosis. In this study, we prepared radiolabeled PS liposomes for detection of vulnerable plaques. Methods: PS liposomes were prepared by lipid film hydration. Phosphatidylcholine (PC) liposomes were prepared as controls. Liposomes (100 or 200 nm) were generated by an extruder to produce PS100, PS200, PC100, and PC200 liposomes. These were then radiolabeled by encapsulating In-111-nitrilotriacetic acid using an active-loading method. In-111 liposomes were incubated with cultured macrophages for 2 h, and the uptake level was measured. For biodistribution studies, the In-111 liposomes were injected intravenously into ddY mice. In addition, the In-111 liposomes were injected into apolipoprotein E-deficient (apoE-/-) mice, and the aortas were harvested for autoradiography and oil red O staining. For SPECT imaging, In-111 liposomes were injected intravenously into Watanabe heritable hyperlipidemic rabbits and scanned 48 h after injection. Results: The radiochemical yields were greater than 95% for all the prepared In-111 liposomes. The level of in vitro uptake by macrophages was 60.5, 14.7, 32.0, and 14.4 percentage injected dose per milligram of protein for In-111-PS100, In-111-PC100, In-111-PS200, and In-111-PC200, respectively. In biodistribution studies, high spleen uptake was seen with PC liposomes. Liver uptake was high for all liposomes but was lowest with In-111-PS200. The blood half-lives were 3.2, 22.0, 3.6, and 7.4 min for In-111-PS100, In-111-PC100, In-111-PS200, and In-111-PC200, respectively. The distribution of In-111-labeled PS liposomes into atherosclerotic regions determined by autoradiography was well matched with the results of oil red O staining in apoE-/- mice. The target-to-nontarget ratios were 2.62, 2.23, 3.27, and 2.51 for In-111-PS100, In-111-PC100, In-111-PS200, and In-111-PC200, respectively. The aorta was successfully visualized by SPECT at 48 h after In-111-labeled PS liposome injection; however, high liver uptake was also observed. Discussion: From the in vitro uptake study, it has been demonstrated that macrophage targeting was accomplished by PS modification. Also, an atherosclerotic region was successfully detected by In-111-PS200 in apoE-/- mice and Watanabe heritable hyperlipidemic rabbits in vivo. Liposome modification to obtain slower blood clearance and lower liver uptake would be required to improve the SPECT images.

    DOI: 10.2967/jnumed.113.123158

    Web of Science

    researchmap

  • Characterization of Cancer Stem-Like Cells Derived from Mouse Induced Pluripotent Stem Cells Transformed by Tumor-Derived Extracellular Vesicles

    Ting Yan, Akifumi Mizutani, Ling Chen, Mai Takaki, Yuki Hiramoto, Shuichi Matsuda, Tsukasa Shigehiro, Tomonari Kasai, Takayuki Kudoh, Hiroshi Murakami, Junko Masuda, Mary J. C. Hendrix, Luigi Strizzi, David S. Salomon, Li Fu, Masaharu Seno

    JOURNAL OF CANCER   5 ( 7 )   572 - 584   2014

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:IVYSPRING INT PUBL  

    Several studies have shown that cancer niche can perform an active role in the regulation of tumor cell maintenance and progression through extracellular vesicles-based intercellular communication. However, it has not been reported whether this vesicle-mediated communication affects the malignant transformation of normal stem cells/progenitors. We have previously reported that the conditioned medium derived from the mouse Lewis Lung Carcinoma (LLC) cell line can convert mouse induced pluripotent stem cells (miPSCs) into cancer stem cells (CSCs), indicating that normal stem cells when placed in an aberrant microenvironment can give rise to functionally active CSCs. Here, we focused on the contribution of tumor-derived extracellular vesicles (tEVs) that are secreted from LLC cells to induce the transformation of miPSCs into CSCs. We isolated tEVs from the conditioned medium of LLC cells, and then the differentiating miPSCs were exposed to tEVs for 4 weeks. The resultant tEV treated cells (miPS-LLCev) expressed Nanog and Oct3/4 proteins comparable to miPSCs. The frequency of sphere formation of the miPS-LLCev cells in suspension culture indicated that the self-renewal capacity of the miPS-LLCev cells was significant. When the miPS-LLCev cells were subcutaneously transplanted into Balb/c nude mice, malignant liposarcomas with extensive angiogenesis developed. miPS-LLCevPT and miPS-LLCevDT, the cells established from primary site and disseminated liposarcomas, respectively, showed their capacities to self-renew and differentiate into adipocytes and endothelial cells. Moreover, we confirmed the secondary liposarcoma development when these cells were transplanted. Taken together, these results indicate that miPS-LLCev cells possess CSC properties. Thus, our current study provides the first evidence that tEVs have the potential to induce CSC properties in normal tissue stem cells/progenitors.

    DOI: 10.7150/jca.8865

    Web of Science

    researchmap

  • Cancer stem cells converted from pluripotent stem cells and the cancerous niche.

    Kasai T, Chen L, Mizutani A, Kudoh T, Murakami H, Fu L, Seno M

    Journal of Stem cells & regenerative medicine   10 ( 1 )   2 - 7   2014

     More details

  • Mouse induced pluripotent stem cell microenvironment generates epithelial-mesenchymal transition in mouse Lewis lung cancer cells

    Ling Chen, Akifumi Mizutani, Tomonari Kasai, Ting Yan, Guoliang Jin, Arun Vaidyanath, Bishoy Y. A. El-Aarag, Yixin Liu, Takayuki Kudoh, David S. Salomon, Li Fu, Masaharu Seno

    AMERICAN JOURNAL OF CANCER RESEARCH   4 ( 1 )   80 - U92   2014

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:E-CENTURY PUBLISHING CORP  

    Induced pluripotent stem (iPS) cells may be a powerful tool in regenerative medicine, but their potential tumorigenicity is a significant challenge for the clinical use of iPS cells. Previously, we succeeded in converting miPS cells into cancer stem cells (CSCs) under the conditions of tumor microenvironment. Both stem cells and tumor cells are profoundly influenced by bi-directional communication with their respective microenvironment, which dictates cell fate determination and behavior. The microenvironment derived from iPS cells has not been well studied. In this paper, we have investigated the effects of secreted factors from Nanog-mouse iPS (miPS) cells on mouse Lewis lung cancer (LLC) cells that are found in the conditioned media. The results demonstrated that miPS cells secrete factors that can convert the epithelia phenotype of LLC cells to a mesenchymal phenotype, and that can promote tumorigenisity, migration and invasion. Furthermore, LLC cells that have been exposed to miPS conditioned medium became resistant to apoptosis. These various biological effects suggest that the miPS microenvironment contain factors that can promote an epithelial-mesenchymal transition (EMT) through an active Snail-MMP axis or by suppressing differentiation in LLC cells.

    Web of Science

    researchmap

  • Chlorotoxin-Fc Fusion Inhibits Release of MMP-2 from Pancreatic Cancer Cells

    Samah El-Ghlban, Tomonari Kasai, Tsukasa Shigehiro, Hong Xia Yin, Sreeja Sekhar, Mikiko Ida, Anna Sanchez, Akifumi Mizutani, Takayuki Kudoh, Hiroshi Murakami, Masaharu Seno

    BIOMED RESEARCH INTERNATIONAL   152659   2014

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:HINDAWI PUBLISHING CORPORATION  

    Chlorotoxin (CTX) is a 36-amino acid peptide derived from Leiurus quinquestriatus (scorpion) venom, which inhibits low-conductance chloride channels in colonic epithelial cells. It has been reported that CTX also binds to matrix metalloproteinase-2 (MMP-2), membrane type-1 MMP, and tissue inhibitor of metalloproteinase-2, as well as CLC-3 chloride ion channels and other proteins. Pancreatic cancer cells require the activation of MMP-2 during invasion and migration. In this study, the fusion protein was generated by joining the CTX peptide to the amino terminus of the human IgG-Fc domain without a hinge domain, the monomeric form of chlorotoxin (M-CTX-Fc). The resulting fusion protein was then used to target pancreatic cancer cells (PANC-1) in vitro. M-CTX-Fc decreased MMP-2 release into the media of PANC-1 cells in a dose-dependent manner. M-CTX-Fc internalization into PANC-1 cells was observed. When the cells were treated with chlorpromazine (CPZ), the internalization of the fusion protein was reduced, implicating a clathrin-dependent internalization mechanism of M-CTX-Fc in PANC-1 cells. Furthermore, M-CTX-Fc clearly exhibited the inhibition of the migration depending on the concentration, but human IgG, as negative control of Fc, was not affected. The M-CTX-Fc may be an effective instrument for targeting pancreatic cancer.

    DOI: 10.1155/2014/152659

    Web of Science

    researchmap

  • Mutual dependence between cancer stem cells and their progenies: the niche created by the progenies is sustaining cancer stem cells.

    Cancer Cell & Microenvironment   1 ( 4 )   141 - 144   2014

     More details

  • Cytokine expression in human dermal fibroblasts stimulated with eosinophil cationic protein measured by protein array

    Takamaro Sato, Yoshihiko Soga, Tomoko Yamaguchi, Michio Meguro, Hiroshi Maeda, Joji Tada, Takayuki Otani, Masaharu Seno, Shogo Takashiba

    ASIAN PACIFIC JOURNAL OF ALLERGY AND IMMUNOLOGY   31 ( 4 )   271 - 276   2013.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ALLERGY IMMUNOL SOC THAILAND,  

    Background: Eosinophil cationic protein (ECP) was reported previously to be involved in allergic inflammation with cytotoxic activity. On the other hand, recent studies showed that ECP did not induce cell death but inhibited the growth of cancer-derived cells. Our previous study indicated that human ECP enhanced differentiation of rat neonatal cardiomyocytes and stress fiber formation in Balb/c 3T3 mouse fibroblasts, while the effects of human ECP on human fibroblasts are unknown.
    Objective: The present study was performed to determine the effects of human ECP on cytokine expression in human fibroblasts by protein array.
    Methods: The effects of recombinant human ECP (rhECP) on normal human dermal fibroblasts (NHDF) were examined by assaying cell growth. Furthermore, cytokine expression of NHDF stimulated by ECP, which could influence cell growth, was evaluated by protein array.
    Results: ECP was not cytotoxic but enhanced the growth of NHDF. The peak rhECP concentration that enhanced the cell counts by 1.56-fold was 100 ng/mL, which was significantly different from cultures without ECP stimulation (ANOVA/ Scheffe's test, P &lt; 0.05). Array analyses indicated that ciliary neurotrophic factor (CNTF), neutrophil-activating peptide (NAP)-2, and neurotrophin (NT)-3 were significantly upregulated in NHDF stimulated with 100 ng/mL ECP compared to those without stimulation.
    Conclusion: ECP is not cytotoxic but enhances the growth of NHDF. CNTF, NAP-2, and NT-3 were suggested to be involved in enhancing the growth of NHDF. These findings will contribute to determination of the role of ECP in allergic inflammation.

    DOI: 10.12932/AP0287.31.4.2013

    Web of Science

    researchmap

  • High epiregulin expression in human U87 glioma cells relies on IRE1 alpha and promotes autocrine growth through EGF receptor

    Gregor Auf, Arnaud Jabouille, Maylis Delugin, Sylvaine Guerit, Raphael Pineau, Sophie North, Natalia Platonova, Marlene Maitre, Alexandre Favereaux, Peter Vajkoczy, Masaharu Seno, Andreas Bikfalvi, Dmitri Minchenko, Oleksandr Minchenko, Michel Moenner

    BMC CANCER   13   13 - 597   2013.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOMED CENTRAL LTD  

    Background: Epidermal growth factor (EGF) receptors contribute to the development of malignant glioma. Here we considered the possible implication of the EGFR ligand epiregulin (EREG) in glioma development in relation to the activity of the unfolded protein response (UPR) sensor IRE1 alpha. We also examined EREG status in several glioblastoma cell lines and in malignant glioma.
    Methods: Expression and biological properties of EREG were analyzed in human glioma cells in vitro and in human tumor xenografts with regard to the presence of ErbB proteins and to the blockade of IRE1 alpha. Inactivation of IRE1 alpha was achieved by using either the dominant-negative strategy or siRNA-mediated knockdown.
    Results: EREG was secreted in high amounts by U87 cells, which also expressed its cognate EGF receptor (ErbB1). A stimulatory autocrine loop mediated by EREG was evidenced by the decrease in cell proliferation using specific blocking antibodies directed against either ErbB1 (cetuximab) or EREG itself. In comparison, anti-ErbB2 antibodies (trastuzumab) had no significant effect. Inhibition of IRE1 alpha dramatically reduced EREG expression both in cell culture and in human xenograft tumor models. The high-expression rate of EREG in U87 cells was therefore linked to IRE1 alpha, although being modestly affected by chemical inducers of the endoplasmic reticulum stress. In addition, IRE1-mediated production of EREG did not depend on IRE1 RNase domain, as neither the selective dominant-negative invalidation of the RNase activity (IRE1 kinase active) nor the siRNA-mediated knockdown of XBP1 had significant effect on EREG expression. Finally, chemical inhibition of c-Jun N-terminal kinases (JNK) using the SP600125 compound reduced the ability of cells to express EREG, demonstrating a link between the growth factor production and JNK activation under the dependence of IRE1 alpha.
    Conclusion: EREG may contribute to glioma progression under the control of IRE1 alpha, as exemplified here by the autocrine proliferation loop mediated in U87 cells by the growth factor through ErbB1.

    DOI: 10.1186/1471-2407-13-597

    Web of Science

    researchmap

  • Development of an Epidermal Growth Factor Derivative with EGFR Blocking Activity Reviewed

    Clara Panosa, Francesc Tebar, Montserrat Ferrer-Batalle, Humphrey Fonge, Masaharu Seno, Raymond M. Reilly, Anna Massaguer, Rafael De Llorens

    PLOS ONE   8 ( 7 )   2013.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PUBLIC LIBRARY SCIENCE  

    The members of the epidermal growth factor (EGF)/ErbB family are prime targets for cancer therapy. However, the therapeutic efficiency of the existing anti-ErbB agents is limited. Thus, identifying new molecules that inactivate the ErbB receptors through novel strategies is an important goal on cancer research. In this study we have developed a shorter form of human EGF (EGFt) with a truncated C-terminal as a novel EGFR inhibitor. EGFt was designed based on the superimposition of the three-dimensional structures of EGF and the Potato Carboxypeptidase Inhibitor (PCI), an EGFR blocker previously described by our group. The peptide was produced in E. coli with a high yield of the correctly folded peptide. EGFt showed specificity and high affinity for EGFR but induced poor EGFR homodimerization and phosphorylation. Interestingly, EGFt promoted EGFR internalization and translocation to the cell nucleus although it did not stimulate the cell growth. In addition, EGFt competed with EGFR native ligands, inhibiting the proliferation of cancer cells. These data indicate that EGFt may be a potential EGFR blocker for cancer therapy. In addition, the lack of EGFR-mediated growth-stimulatory activity makes EGFt an excellent delivery agent to target toxins to tumours over-expressing EGFR.

    DOI: 10.1371/journal.pone.0069325

    Web of Science

    researchmap

  • Nano-micrometer-architectural acidic silica prepared from iron oxide of Leptothrix ochracea origin. Reviewed

    Hashimoto H, Itadani A, Kudoh T, Fukui S, Kuroda Y, Seno M, Kusano Y, Ikeda Y, Benino Y, Nanba T, Nakanishi M, Fujii T, Takada J.

    ACS Appl Mater Interfaces   12 ( 5 )   518 - 523   2013.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • A novel remote loading method with solubility gradient to encapsulate effevtive amount of taxanes into liposomes.

    Tsukasa Shigehiro, Tomonari Kasai, Akifumi Mizutani, Hiroshi Murakami, Katsuhiko Mikuni, Tadakatsu Mandai, Hiroki Hamada, Masaharu Seno

    CANCER RESEARCH   73 ( 8 )   2013.4

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2013-LB-8

    Web of Science

    researchmap

  • マウスiPS細胞から作るがん幹細胞モデル

    笠井智成, 陳 凌, 工藤孝幸, 水谷昭文, 妹尾昌治

    細胞工学   32 ( 3 )   330 - 337   2013

     More details

  • Development and characterization of cancer stem cell model from mouse iPS cells

    Ling Chen, Shuichi Matsuda, Tomonari Kasai, Yuh Sugii, Masashi Okada, Koichi Igarashi, Ayano Satoh, Takayuki Kudoh, Takayuki Kudoh, Li Fu, Masaharu Seno

    CANCER RESEARCH   72   2012.4

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2012-418

    Web of Science

    researchmap

  • Preparation and evaluation of glycosylated paclitaxel loaded in tastuzumab-immunoliposome

    Masaharu Murakami, Tomonari Kasai, Masashi Okada, Katsuhiko Mikuni, Naoyoshi Egashira, Masaharu Seno, Hiroki Hamada

    CANCER RESEARCH   72   2012.4

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2012-5700

    Web of Science

    researchmap

  • Chemo-enzymatic synthesis of glycolyl-ester-linked taxol-monosaccharide conjugate and its drug delivery system using hepatitis B virus envelope L bio-nanocapsules. International journal

    Kei Shimoda, Manabu Hamada, Masaharu Seno, Tadakatsu Mandai, Hiroki Hamada

    Biochemistry insights   5   11 - 5   2012

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Chemo-enzymatic synthesis of glycolyl-ester-linked taxol-glucose conjugate, ie, 7-glycolyltaxol 2″-O-α-D-glucoside, was achieved by using α-glucosidase as a biocatalyst. The water-solubility of 7-glycolyltaxol 2″-O-α-D-glucoside (21 μM) was 53 fold higher than that of taxol. The hepatitis B virus envelope L particles (bio-nanocapsules) are effective for delivering 7-glycolyltaxol 2″-O-α-D-glucoside to human hepatocellular carcinoma NuE cells.

    DOI: 10.4137/BCI.S9824

    PubMed

    researchmap

  • Extracellular Matrix Modulates Insulin Production During Differentiation of AR42J Cells: Functional Role of Pax6 Transcription Factor

    Kohei Hamamoto, Satoko Yamada, Akemi Hara, Tsutomu Kodera, Masaharu Seno, Itaru Kojima

    JOURNAL OF CELLULAR BIOCHEMISTRY   112 ( 1 )   318 - 329   2011.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-BLACKWELL PUBLISHING, INC  

    Extracellular matrix (ECM) modulates differentiation of pancreatic beta-cells during development. However, the mechanism by which ECM proteins modulate differentiation is not totally clear. We investigated the effect of ECM proteins on differentiation beta-cells in vitro. We investigated the effect of basement membrane ECM on differentiation of AR42J cells and rat ductal cells. First, we examined the effect of reconstituted basement membrane, Matrigel on differentiation of AR42J cells induced by activin and betacellulin. Matrigel augmented insulin production and increased the expression of GLUT2, SUR1, and glucokinase. Among various transcription factors investigated, Matrigel markedly upregulated the expression of Pax6. When Pax6 was overexpressed in cells treated with activin and betacellulin, the expression of insulin was upregulated. Conversely, knockdown of Pax6 significantly reduced the insulin expression in cells cultured on Matrigel. The effects of Matrigel on insulin-production and induction of Pax6 were reproduced partially by laminin-1, a major component of Matrigel, and inhibited by anti-integrin-beta 1 antibody. Matrigel also enhanced the activation of p38 mitogen-activated kinase induced by activin and betacellulin, which was inhibited by anti-beta 1 antibody. Finally, the effect of Matrigel on differentiation was reproduced in rat cultured ductal cells, and Matrigel also increased the expression of Pax6. These results indicate that basement membrane ECM augments differentiation of pancreatic progenitor cells to insulin-secreting cells by upregulating the expression of Pax6. J. Cell. Biochem. 112: 318329, 2011. (C) 2010 Wiley-Liss, Inc.

    DOI: 10.1002/jcb.22930

    Web of Science

    researchmap

  • Construction of a high-efficiency multi-site-directed mutagenesis

    Haidong Tan, Yueguang Li, Ling Chen, Tomonari Kasai, Masaharu Seno

    AFRICAN JOURNAL OF BIOTECHNOLOGY   10 ( 3 )   449 - 452   2011.1

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC JOURNALS  

    Although site-directed mutagenesis has been used in many fields, it still has low rate of success and high cost because of low-yield target products. A modified method for multi-site-directed mutagenesis was developed with shifted primer design and cold-start polymerase chain reaction (PCR). The developed method was successfully applied to hexapeptide gene synthesis and recombinant enterokinase gene modification in the plasmids pET41a and pET24b-EK. The efficiency was pronounced at a 1:10 molar ratio of 7-base mutant products to 705-bp fragment products as control. Even in a 10-base substitution mutagenic PCR, a 1:50 molar ratio of mutant products to 705-bp fragment products was reached. Meanwhile, the quality of mutants was proved through the transformation efficiency and sequencing. This method was beneficial to prepare high-quality multibase mutagenesis and also implied that large-scale multibase mutagenesis was feasible, efficient, economical, and productive.

    Web of Science

    researchmap

  • Generation of Cancer Stem Cell Model from Mouse iPS Cells.

    A. Mizutani, S-I. Matsuda, T. Kasai, T. Kudoh, L. Chen, M. Seno

    MOLECULAR BIOLOGY OF THE CELL   22   2011

     More details

    Language:English   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Analysis of endothelium mimicry by the CSC-like cells derived from iPS cells.

    S-I. Matsuda, A. Mizutani, T. Kasai, A. Satoh, T. Kudoh, L. Chen, M. Seno

    MOLECULAR BIOLOGY OF THE CELL   22   2011

     More details

    Language:English   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Optimization of Bacterial Plasmid Transformation Using Nanomaterials Based on the Yoshida Effect

    Haidong Tan, Li Fu, Masaharu Seno

    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   11 ( 12 )   4962 - 4973   2010.12

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    With the help of sepiolite, a unique method for transforming DNA into bacteria, based on the Yoshida effect, has been developed recently. However, we confronted many problems when this newest method was tried. Only a few transformants could be obtained even when 100 ng of plasmid pET15b was used, and a successful result seemed difficult to repeat. To address this problem, we optimized the operating method and could achieve about 15,000 transformants using the same amount of plasmid, which could match the efficiency gained using the calcium chloride transformation method. Meanwhile, the results could also be reproduced well. In the same way, carbon nanotubes were used to attain more than 15,000 transformants in the same situation. Therefore, the transformation method could be extended to other nanomaterials. Meanwhile, compared with the mechanism previously reported, we verified quite a different principle for the mechanism responsible for such a transformation. In sum, this unique transformation can be developed to become the third widely-used transformation method in laboratories in addition to the chemical method and electroporation.

    DOI: 10.3390/ijms11124962

    Web of Science

    researchmap

  • First Report of Cucumber mosaic virus in Sweet Cherry in the People&apos;s Republic of China

    H. D. Tan, S. Y. Li, X. F. Du, M. Seno

    PLANT DISEASE   94 ( 11 )   1378 - 1378   2010.11

     More details

    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER PHYTOPATHOLOGICAL SOC  

    DOI: 10.1094/PDIS-07-10-0549

    Web of Science

    researchmap

  • Novel and simple loading procedure of cisplatin into liposomes and targeting tumor endothelial cells

    M. Hirai, H. Minematsu, Y. Hiramatsu, H. Kitagawa, T. Otani, S. Iwashita, T. Kudoh, L. Chen, Y. Li, M. Okada, D. S. Salomon, K. Igarashi, M. Chikuma, M. Seno

    INTERNATIONAL JOURNAL OF PHARMACEUTICS   391 ( 1-2 )   274 - 283   2010.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Although intravenous administration of high levels of cisplatin (CDDP) are limited due to its severe side effects, efficient delivery of CDDP directly to the tumor should improve the therapeutic response while potentially by-passing significant side effects.
    High loading of CDDP into liposomes is one technique that could be used as a potential drug delivery system. Since cis-diamminedinitratoplatinum (CDDP3) is highly soluble in water and converts to CDDP in the presence of chloride ions, we encapsulated CDDP3 into liposomes in the absence of chloride ions and supplemented chloride ions to prepare CDDP-encapsulated liposomes (CDDP-Lip) resulting in a significantly improved loading efficiency of COOP. We further conjugated the CDDP-Lip with Sialyl Lewis(X) (CDDP-SLX-Lip) because we previously demonstrated Sialyl Lewis(X) enhanced efficient accumulation of liposomes into tumors in vivo. CDDP-SLX-Lip treated mice showed a survival rate of 75% at 14 days even if a lethal level of CDDP was injected into mice. Loss of body weight was negligible and no histological abnormality was found in a variety of normal tissues. Accumulation of CDDP-SLX-Lip was about 6 times more than that of CDDP-Lip or CDDP. As the result, there was better antitumor activity of CDDP-SLX-Lip than that of CDDP-Lip with significantly less toxic effects in normal tissues. (C) 2010 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.ijpharm.2010.02.030

    Web of Science

    researchmap

  • E-selectin targeting to visualize tumors in vivo

    Masahiko Hirai, Yoshie Hiramatsu, Shinki Iwashita, Takayuki Otani, Ling Chen, Yue-guang Li, Masashi Okada, Kazunori Oie, Koich Igarashi, Hideaki Wakita, Masaharu Seno

    CONTRAST MEDIA & MOLECULAR IMAGING   5 ( 2 )   70 - 77   2010.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JOHN WILEY & SONS LTD  

    Generally angiogenic factors induce the expression of E-selectin in vascular endothelial cells in the tumors. In this study, we employed an anti-E-selectin monoclonal antibody to target tumors in vivo and evaluated an optical imaging reagent to visualize tumor regions. The anti-E-selectin antibody was conjugated on the surface of liposomes, which encapsulated the near-infrared fluorescent substances Cy3 or Cy5.5. The liposomes efficiently recognized human umbilical vein endothelial cells only when E-selectin was induced by angiogenic factors such as TNF-alpha in vitro. Cy5.5 encapsulated into liposomes that were conjugated with an anti-E-selectin antibody successfully visualized Ehrlich ascites tumor cells when transplanted into mice. Thus, E-selectin targeting with liposomes containing a near-infrared fluorescent dye was found effective in visualizing tumors in vivo. This strategy should be extremely useful as a method to identify sentinel lymphatic nodes and angiogenic tumors as well as use for drug delivery to tumor cells. Copyright (C) 2010 John Wiley & Sons, Ltd.

    DOI: 10.1002/cmmi.367

    Web of Science

    researchmap

  • Production of biologically active IgG hinge-tag soluble epidermal growth factor receptors (ErbB)

    Takayuki Otani, Toshihiro Hashizume, Tadahiro Nagaoka, Tomoko Fukuda, Careen K. Tang, David S. Salomon, Masaharu Seno

    BIOTECHNOLOGY LETTERS   32 ( 3 )   361 - 366   2010.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER  

    The extracellular domains (ECD) of epidermal growth factor receptors, ErbB1, 2, 3 and 4, were designed as soluble dimeric forms. Each ECD was fused to a short hinge region derived from IgG, such that the stable dimer could be formed with disulfide bridges. This hinge-tagged design minimized the molecular weight to approximately 50% of the conventional Fc-fusion design without an Fc domain of IgG. The refolded dimers could be easily analyzed and characterized by SDS-PAGE. Hinge-tagged soluble ErbBs demonstrated significant affinity for betacellulin and heregulin. The IgG hinge-tag should be a simple method to design soluble dimers that would be useful for high throughput screening of ligands, antagonists or derivatives.

    DOI: 10.1007/s10529-009-0160-9

    Web of Science

    researchmap

  • Administration of Conophylline and Betacellulin-delta 4 Increases the beta-cell Mass in Neonatal Streptozotocin-treated Rats

    Tsutomu Kodera, Satoko Yamada, Yoritsuna Yamamoto, Akemi Hara, Yuji Tanaka, Masaharu Seno, Kazuo Umezawa, Izumi Takei, Itaru Kojima

    ENDOCRINE JOURNAL   56 ( 6 )   799 - 806   2009.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN ENDOCRINE SOC  

    The present study was conducted to examine the effect of administration of conophylline (CnP) and betacellulin delta 4 (BTC delta 4) on the beta-cell mass in neonatal streptozotocin-treated rats (neonatal STZ rats). STZ (100 mu g/g) was injected into neonatal rats, and then CnP (2 mu g/g) and/or BTC delta 4 (200 pmol/g) were administered to neonatal STZ rats for 1 week. The plasma glucose concentration was monitored, and an intraperitoneal glucose tolerance test (ipGTT) was performed on day 8 and at 8 weeks after the STZ injection. In neonatal STZ rats treated with control solution (S group), the plasma glucose concentration increased for several days after the STZ injection, returned to nearly normal levels, and then increased gradually after six weeks of age. Eight weeks after the STZ-injection, the plasma glucose concentration was increased significantly compared to that of normal rats. The glucose response to ipGTT was significantly reduced in neonatal STZ rats treated with CnP (CnP group), BTC delta 4 (delta 4 group) and CnP+BTC delta 4 (CnP+delta 4 group). The beta-cell mass and the insulin content of the pancreas were significantly increased in the CnP group and delta 4 group. The effect of CnP+delta 4 was greater than ;that of CnP alone or BTC delta 4 alone. CnP+BTC delta 4 significantly increased the number of PDX-1-positive ductal cells and the number of insulin/BrdU double-positive ductal cells. These results indicate the efficacy of CnP and BTC delta 4 in increasing the beta-cells mass of neonatal STZ-treated rats.

    DOI: 10.1507/endocrj.K09E-158

    Web of Science

    researchmap

  • Delivery of sodium borocaptate to glioma cells using immunoliposome conjugated with anti-EGFR antibodies by ZZ-His

    Bin Feng, Kazuhito Tomizawa, Hiroyuki Michiue, Shin-ichi Miyatake, Xiao-Jian Han, Atsushi Fujimura, Masaharu Seno, Mitsunori Kirihata, Hideki Matsui

    BIOMATERIALS   30 ( 9 )   1746 - 1755   2009.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCI LTD  

    Nanoparticles are effective of delivering cargo into cells. Here, sodium borocaptate (BSH) was encapsulated in liposomes composed of nickel lipid, and anti-epidermal growth factor receptor (EGFR) antibodies were conjugated to the liposomes using the antibody affinity motif of protein A (ZZ) as an adaptor (immunoliposomes). The immunoliposomes were used to deliver BSH into EGFR-overexpressing glioma cells. Immunohistochemical analysis using an anti-BSH monoclonal antibody revealed that BSH was delivered effectively into the cells but not into EGFR-deficient glioma or primary astrocytes. In an animal model of brain tumors, both the liposomes and the BSH were only observed in the tumor. Moreover, the efficiency of (10)B&apos;s delivery into glioma cells was confirmed by inductively coupled plasma-atomic emission spectrometry (ICP-AES) both in vitro and in vivo. The results suggest that this system utilizing immunoliposomes provides an effective means of delivering (10)B into glioma cells in boron neutron capture therapy (BNCT). (C) 2008 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.biomaterials.2008.12.010

    Web of Science

    researchmap

  • Human eosinophil cationic protein enhances stress fiber formation in Balb/c 3T3 fibroblasts and differentiation of rat neonatal cardiomyocytes

    Takayuki Fukuda, Miki Iwata, Midori Kitazoe, Takashi Maeda, David Salomon, Satoshi Hirohata, Katsuyuki Tanizawa, Shun&apos;ichi Kuroda, Masaharu Seno

    GROWTH FACTORS   27 ( 4 )   228 - 236   2009

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    We found that eosinophil cationic protein (ECP) stimulated the growth of mouse Balb/c 3T3 fibroblasts. ECP-treated 3T3 cells were more flattened and exhibited enhanced stress fiber formation. The enhancement of cytoskeleton after addition of recombinant ECP appeared stable and was able to inhibit disassembly of actin filaments that was induced by fibroblast growth factor-2. The ROCK inhibitor, Y-27632, abrogated this enhancement on stress fiber formation that was induced by ECP indicating the involvement of Rho/ROCK signaling pathway. The effect of ECP was assessed on the differentiation of primary cardiomyocytes derived from rat neonatal heart since the development of actin filaments is significantly related with organization of stress fibers. As the result, both beating rate and the expression of cardiac muscle specific markers such as atrial natriuretic factor were enhanced in the presence of ECP. Thus ECP may also function as a cardiomyocyte differentiation factor.

    DOI: 10.1080/08977190902987149

    Web of Science

    researchmap

  • Spherical self-organizing map as a helpful tool to identify category-specific cell surface markers

    Tuoya, Yuh Sugii, Hitomi Satoh, Dongwei Yu, Yasaburo Matsuura, Heizo Tokutaka, Masaharu Seno

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   376 ( 2 )   414 - 418   2008.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We analyzed gene expression profiles of five tumor cell lines (NB2a, NB41A3, C1300N18, BC3H1, and Neuro2a) derived from a category of nervous system using our originally developed cell surface marker DNA microarray in order to search for tumor-specific cell surface markers common to these cells. To visualize the expression patterns and to extract candidate genes of interest based on the expression profiles of several cell lines, we employed the clustering procedure of spherical self-organizing-map. As the result, three candidates of tumor-specific cell surface markers were picked up when the expression profiles were compared with that from normal brain tissue. RT-qPCR showed the expression of these genes was higher in tumor cells than in normal brain. Here we demonstrated the spherical self-organizing-map analysis should be useful to identify the candidates of cell surface markers common and specific to the group of cells or tissues of interest. (C) 2008 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2008.09.010

    Web of Science

    researchmap

  • Intracellular delivery of glutathione S-transferase-fused proteins into mammalian cells by polyethylenimine-glutathione conjugates

    Hitoshi Murata, Junichiro Futami, Midori Kitazoe, Takayuki Yonehara, Hidetaka Nakanishi, Megumi Kosaka, Hiroko Tada, Masakiyo Sakaguchi, Yasuyuki Yagi, Masaharu Seno, Nam-ho Huh, Hidenori Yamada

    JOURNAL OF BIOCHEMISTRY   144 ( 4 )   447 - 455   2008.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OXFORD UNIV PRESS  

    The glutathione S-transferase (GST)-fused protein expression system has been extensively used to generate a large quantity of proteins and has served for functional analysis in vitro. In this study, we developed a novel approach for the efficient intracellular delivery of GST-fused proteins into living cells to expand their usefulness up to in vivo use. Since protein cationization techniques are powerful strategies for efficient intracellular uptake by adsorptive-mediated endocytosis, GST-fused proteins were cationized by forming a complex with a polycationic polyethylenimine (PEI)-glutathione conjugate. On screening of protein transduction, optimized PEI-glutathione conjugate for protein transduction was characterized by a partly oligomerized mixture of PEI with average molecular masses of 600 (PEI600) modified with multiple glutathiones, which could have sufficient avidity for GST. Furthermore, enhanced endosomal escape of transduced GST-fused proteins was observed when they were delivered with a glutathione-conjugated PEI600 derivative possessing a hydroxybutenyl moiety. These results were confirmed by both intracellular confocal imaging of GST-fused green fluorescent protein and activation of an endogenous growth signal transduction pathway by a GST-fused constitutively active mutant of a kinase protein. These PEI-glutathione conjugates seem to be convenient molecular tools for protein transduction of widely used GST-fused proteins.

    DOI: 10.1093/jb/mvn087

    Web of Science

    researchmap

  • Smad2 functions as a co-activator of canonical Wnt/beta-catenin signaling pathway independent of Smad4 through histone acetyltransferase activity of p300

    Morihisa Hirota, Kazuhide Watanabe, Shin Hamada, Youping Sun, Luigi Strizzi, Mario Mancino, Tadahiro Nagaoka, Monica Gonzales, Masaharu Seno, Caterina Bianco, David S. Salomon

    CELLULAR SIGNALLING   20 ( 9 )   1632 - 1641   2008.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE INC  

    Both canonical Wnt/beta-catenin and TGF beta/Smad signaling pathways coordinately regulate pattern formation during embryogenesis as well as tumor progression. Evidence of cross-talk between these two pathways has been reported. Here we demonstrated that the Activin-like kinase 4 (Alk4)/Smad2 pathway facilitates the transcriptional activity of the oncogenic Wnt/beta-catenin/Tcf4 pathway through a novel Smad4-independent mechanism. Upon activation, Smad2 physically interacted with Tcf4, beta-catenin and the co-activator p300 to enhance transcriptional activity of beta-catenin/Tcf4 through the histone acetyltransferase activity of p300. Transactivation by Smad2 was independent of a Smad-binding element (SBE) and Smad4. Indeed, the enhancement of beta-catenin/Tcf4 transcriptional activity by activated Smad2 was negatively regulated by the presence of Smad4. Moreover, a tumor-derived missense mutant of Smad2, lacking the ability to bind to Smad4 was still able to enhance the Tcf4 transcriptional reporter in the presence of p-catenin and Tcf4. Our findings suggest that Smad2 may function as an activator of canonical Wnt/beta-catenin/Tcf4 signaling through a SBE/Smad4-independent pathway. Published by Elsevier Inc.

    DOI: 10.1016/j.cellsig.2008.05.003

    Web of Science

    researchmap

  • Reversal of streptozotocin-induced hyperglycemia by continuous supply of betacellulin in mice

    Yoritsuna Yamamoto, Satoko Yamada, Tsutomu Kodera, Akemi Hara, Kazuo Motoyoshi, Yuji Tanaka, Tadahiro Nagaoka, Masaharu Seno, Itaru Kojima

    GROWTH FACTORS   26 ( 4 )   173 - 179   2008.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:TAYLOR & FRANCIS LTD  

    Previous studies have shown the efficacy of betacellulin (BTC) to promote P-cell regeneration. Because of its short half-life, however, the effect of BTC may have been underestimated. This study was conducted to assess the effect of continuous administration of BTC on P-cell regeneration. Adenovirus vectors encoding proBTC (Ad-proBTC) and mature BTC (Ad-mBTC) were prepared, and the efficacy of secretion of BTC was compared in AML12 hepatocytes. When AML12 cells were infected with Ad-proBTC or Ad-mBTC, cells infected with Ad-mBTC secreted considerably larger amount of BTC. We then infused Ad-mBTC into the mouse tail vein. Expression of BTC was detected in the liver for at least 21 days, and serum BTC was maintained at approximately 1 ng/ml for 7 days. When Ad-mBTC was infused immediately after administration of STZ (170 mg/kg), elevation of the plasma glucose induced by STZ was markedly inhibited, and the plasma glucose concentration remained at less than 200 mg/dl for 21 days. The insulin content and the P-cell mass were significantly increased in Ad-mBTC-infused mice. These results indicate that continuous administration of BTC is quite effective in promoting regeneration of P-cells.

    DOI: 10.1080/08977190802136854

    Web of Science

    researchmap

  • Cell type dependent endocytic internalization of ErbB2 with an artificial peptide ligand that binds to ErbB2

    Toshihiro Hashizume, Takayuki Fukuda, Tadahiro Nagaoka, Hiroko Tada, Hidenori Yamada, Kazuhide Watanabe, David S. Salomon, Masaharu Seno

    CELL BIOLOGY INTERNATIONAL   32 ( 7 )   814 - 826   2008.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD  

    ErbB2, which is a member of the epidermal growth factor (erbB) receptor family, is frequently overexpressed in breast and ovarian cancers. Antibody and small molecule anti-tyrosine kinase inhibitors have been developed for targeted therapies for cancers overexpressing erbB2. Internalization and downregulation of erbB2, which is induced by a ligand, may be important for efficacious therapeutic effects. However, ligand-dependent erbB2 internalization has not been well characterized. Here we investigated the internalization of erbB2 in SKBr3 and SKOv3 cells, both overexpressing erbB2, using an EC-1 peptide fused to eGFP (EC-eGFP), which specifically binds to erbB2. ErbB2 was internalized in SKOv3 cells when the cells were treated with EC-eGFP. The accumulation of endosomal erbB2 was EC-eGFP dependent, which colocalized with transferrin implying endocytosis via clathrin-coated pits. In contrast, internalization of erbB2 was not observed in SKBr3 cells. As a result, two different mechanisms, which are cell type dependent for the internalization of erbB2, are proposed. (C) 2008 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.cellbi.2008.03.012

    Web of Science

    researchmap

  • A betacellulin mutant promotes differentiation of pancreatic acinar AR42J cells into insulin-producing cells with low affinity of binding to ErbB1

    Tadahiro Nagaoka, Takayuki Fukuda, Toshihiro Hlashizume, Tomoko Nishiyama, Hiroko Tada, Hidenori Yamada, David S. Salomon, Satoko Yamada, Itaru Kojima, Masaharu Seno

    JOURNAL OF MOLECULAR BIOLOGY   380 ( 1 )   83 - 94   2008.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD  

    Betacellulin (BTC) is one of the members of the epidermal growth factor (EGF) ligand family of ErbB receptor tyrosine kinases. It is a differentiation factor as well as a potent mitogen. BTC promotes the differentiation of pancreatic acinar-derived AR42J cells into insulin-producing cells. It independently and preferentially binds to two type I tyrosine kinase receptors, the EGF receptor (ErbB1) and ErbB4. However, the physiochemical characteristics of BTC that are responsible for its preferential binding to these two receptors have not been fully defined. In this study, to investigate the essential amino acid residues of BTC for binding to the two receptors, we introduced point mutations into the EGF domain of BTC employing error-prone PCR. The receptor binding abilities of 190 mutants expressed in Escherichia coli were assessed by enzyme immunoassay. Replacement of the glutamic acid residue at position 88 with a lysine residue in BTC was found to produce a significant loss of affinity for binding to ErbB1, while the affinity of binding to ErbB4 was unchanged. In addition, the mutant of BTC-E/88/K showed less growth-promoting activity on BALB/c 3T3 cells compared with that of the wild-type BTC protein. Interestingly, the BTC mutant protein promoted differentiation of pancreatic acinar AR42J cells at a high frequency into insulin-producing cells compared with AR42J cells that were treated with wild-type BTC protein. These results indicate the possibility of designing BTC mutants, which have an activity of inducing differentiation only, without facilitating growth promotion. (C) 2008 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.jmb.2008.03.054

    Web of Science

    researchmap

  • A simple method to induce differentiation of murine bone marrow mesenchymal cells to insulin-producing cells using conophylline and betacellulin-delta4

    Etsuko Hisanaga, Kee-Yong Park, Satoko Yamada, Hiromi Hashimoto, Toshlyukj Takeuchi, Masatomo Mori, Masaharu Seno, Kazuo Umezawa, Izumi Takei, Itaru Kojima

    ENDOCRINE JOURNAL   55 ( 3 )   535 - 543   2008.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN ENDOCRINE SOC  

    The present study was conducted to establish a method to induce differentiation of bone marrow (MB)-derived mesenchymal cells into insulin-producing cells. When mouse BM-derived mesenchymal cells were cultured for 60 days in medium containing 10% fetal calf serum and 25 mM glucose, they expressed insulin. Addition of activin A and betacellulin (BTC) accelerated differentiation, and immunoreactive insulin was detected 14 days after the treatment. Insulin-containing secretory granules were observed in differentiated cells by electron microscopy. Treatment of BM-derived mesenchymal cells with conophylline (CnP) and BTC-delta4 further accelerated differentiation, and mRNA for insulin was detected 5 to 7 days after the treatment. Mesencymal cells treated with CnP and BTC-delta4 responded to a high concentration of glucose and secreted mature insulin. When these cells were transplanted into streptozotocin-treated mice, they markedly reduced the plasma glucose concentration, and the effect continued for at least 4 weeks. These results indicate an efficacy of the combination of CnP and BTC-delta4 in inducing differentiation of BM-derived mesenchymal cells into insulin-producing cells.

    DOI: 10.1507/endocrj.K07E-173

    Web of Science

    researchmap

  • Involvement of MAPK signaling molecules and Runx2 in the NELL1-induced osteoblastic differentiation

    Nobuyuki Bokui, Takayuki Otani, Koichi Igarashi, Junichiro Kaku, Mitsuo Oda, Tadahiro Nagaoka, Masaharu Seno, Kenji Taternatsu, Toshihide Okajima, Takashi Matsuzaki, Kang Ting, Katsuyuki Tanizawa, Shunichi Kuroda

    FEBS LETTERS   582 ( 2 )   365 - 371   2008.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    NELL1 is an extracellular protein inducing osteogenic differentiation and bone formation of osteoblastic cells. To elucidate the intracellular signaling cascade evoked by NELL1, we have shown that NELL1 protein transiently activates the MAPK signaling cascade, induces the phosphorylation of Runx2, and promotes the rapid intracellular accumulation of Tyr-phosphorylated proteins. Unlike BMP2, NELL1 protein does not activate the Smad signaling cascade. These findings suggest that upon binding to a specific receptor NELL1 transduces an osteogenic signal through activation of certain Tyr-kinases associated with the Ras-MAPK cascade, and finally leads to the osteogenic differentiation. (C) 2007 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.febslet.2007.12.006

    Web of Science

    researchmap

  • Morphological and Microstructural Study of Iron Oxide Microtubes formed by Iron Oxidizing Bacteria, Leptothrix ochracea

    H. Hashimoto, H. Asaoka, J. Takada, T. Fujii, M. Nakanishi, S. Yokoyama, R. Murakami, Y. Kusano, Y. Ikeda, M. Seno

    Global Roadmap of Ceramics - ICC2 Proceedings   6-P-011-1-6-P-011-4   2008

     More details

  • Growth factor induction of cripto-1 shedding by glycosylphosphatidylinositol-phospholipase D and enhancement of endothelial cell migration

    Kazuhide Watanabe, Caterina Bianco, Luigi Strizzi, Shin Hamada, Mario Mancino, Veronique Bailly, Wenjun Mo, Dingyi Wen, Konrad Miatkowski, Monica Gonzales, Michele Sanicola, Masaharu Seno, David S. Salomon

    JOURNAL OF BIOLOGICAL CHEMISTRY   282 ( 43 )   31643 - 31655   2007.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Cripto-1 (CR-1) is a glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that has been shown to play an important role in embryogenesis and cellular transformation. CR-1 is reported to function as a membrane-bound co-receptor and as a soluble ligand. Although a number of studies implicate the role of CR-1 as a soluble ligand in tumor progression, it is unclear how transition from the membrane-bound to the soluble form is physiologically regulated and whether differences in biological activity exist between these forms. Here, we demonstrate that CR-1 protein is secreted from tumor cells into the conditioned medium after treatment with serum, epidermal growth factor, or lysophosphatidic acid, and this soluble form of CR-1 exhibits the ability to promote endothelial cell migration as a paracrine chemoattractant. On the other hand, membranebound CR-1 can stimulate endothelial cell sprouting through direct cell-cell interaction. Shedding of CR-1 occurs at the GPI-anchorage site by the activity of GPI-phospholipase D (GPIPLD), because CR-1 shedding was suppressed by siRNA knockdown of GPI-PLD and enhanced by overexpression of GPI-PLD. These findings describe a novel molecular mechanism of CR-1 shedding, which may contribute to endothelial cell migration and possibly tumor angiogenesis.

    DOI: 10.1074/jbc.M702713200

    Web of Science

    researchmap

  • Development of bionanocapsules targeting brain tumors

    Yumi Tsutsui, Kazuhito Tomizawa, Mana Nagita, Hiroyuki Michiue, Tei-ichi Nishiki, Iori Ohmori, Masaharu Seno, Hideki Matsui

    JOURNAL OF CONTROLLED RELEASE   122 ( 2 )   159 - 164   2007.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER  

    Bionanocapsules (BNCs) are hollow nanoparticles that are composed of L protein (the hepatitis B virus surface antigen) and show specific affinity for human hepatocytes. The pre-S1 peptide displayed on the surface of BNCs is the specific ligand for binding to the receptor on human hepatocytes. Therefore, BNCs are not delivered to other tissues, such as the brain. The aim of the present study was to develop a novel drug delivery system (DIDS) targeting brain tumors using BNCs that selectively targeted brain tumors. Epidermal growth factor receptor (EGFR), especially a constitutively active genomic sequence deletion variant of EGFR (EGFRvIII), is overexpressed in human glioblastoma. In the present study, we replaced the pre-S1 peptide with the antibody affinity motif of protein A and made hybrid BNCs conjugated with anti-human EGFR antibody recognizing EGFRvIII. The hybrid BNCs were efficiently delivered to glioma cells but not normal glial cells. Moreover, we confirmed the specific delivery of the hybrid BNCs to brain tumors in an in vivo brain tumor model. These results suggest that this new approach using BNCs is a promising system for brain tumor-targeted drug delivery. (C) 2007 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jconrel.2007.06.019

    Web of Science

    researchmap

  • 'Crystal lattice engineering,' an approach to engineer protein crystal contacts by creating intermolecular symmetry: Crystallization and structure determination of a mutant human RNase 1 with a hydrophobic interface of leucines

    Hidenori Yamada, Taro Tamada, Megumi Kosaka, Kohei Miyata, Shinya Fujiki, Masaru Tano, Masayuki Moriya, Mamoru Yamanishi, Eijiro Honjo, Hiroko Tada, Takeshi Ino, Hiroshi Yamaguchi, Junichiro Futami, Masaharu Seno, Takashi Nomoto, Tomoko Hirata, Motonobu Yoshimura, Ryota Kuroki

    PROTEIN SCIENCE   16 ( 7 )   1389 - 1397   2007.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT  

    A protein crystal lattice consists of surface contact regions, where the interactions of specific groups play a key role in stabilizing the regular arrangement of the protein molecules. In an attempt to control protein incorporation in a crystal lattice, a leucine zipper-like hydrophobic interface (comprising four leucine residues) was introduced into a helical region (helix 2) of the human pancreatic ribonuclease 1 (RNase 1) that was predicted to form a suitable crystallization interface. Although crystallization of wild-type RNase 1 has not yet been reported, the RNase 1 mutant having four leucines (4L-RNase 1) was successfully crystallized under several different conditions. The crystal structures were subsequently determined by Xray crystallography by molecular replacement using the structure of bovine RNase A. The overall structure of 4L-RNase 1 is quite similar to that of the bovine RNase A, and the introduced leucine residues formed the designed crystal interface. To characterize the role of the introduced leucine residues in crystallization of RNase 1 further, the number of leucines was reduced to three or two (3L-and 2L-RNase 1, respectively). Both mutants crystallized and a similar hydrophobic interface as in 4L-RNase 1 was observed. A related approach to engineer crystal contacts at helix 3 of RNase 1 (N4L- RNase 1) was also evaluated. N4L- RNase 1 also successfully crystallized and formed the expected hydrophobic packing interface. These results suggest that appropriate introduction of a leucine zipper-like hydrophobic interface can promote intermolecular symmetry for more efficient protein crystallization in crystal lattice engineering efforts.

    DOI: 10.1110/ps.072851407

    Web of Science

    researchmap

  • Conophylline and betacellulin-delta 4: an effective combination of differentiation factors for pancreatic beta cells Reviewed

    Ryu-ichi Kitamura, Takeki Ogata, Yuji Tanaka, Kazuo Motoyoshi, Masaharu Seno, Izumi Takei, Kazuo Umezawa, Itaru Kojima

    ENDOCRINE JOURNAL   54 ( 2 )   255 - 264   2007.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN ENDOCRINE SOC  

    Conophylline and betacellulin-delta 4 reproduce differentiation-inducing activity of activin A and betacellulin, respectively. We examined the effect of conophylline and betacellulin-delta 4 on P cell differentiation. In AR42J cells, conophylline and betacellulin-delta 4 converted them into insulin-producing cells. Cells treated with conophylline and betacellulin-delta 4 continued to grow after differentiation. Thus, cell number and insulin content were much greater compared to cells treated with activin A and betacellulin. Furthermore, cells treated with conophylline and betacellulin-delta 4 secreted insulin in response to glucose. Likewise, conophylline and betacellulin-delta 4 converted pancreatic ductal cells into insulin-producing cells. Insulin content, cell number and glucose-evoked insulin secretion were significantly greater than those in cells treated with activin A and betacellulin. Transplantation of pseudoislets prepared using ductal cells treated with conophylline and betacellulin-delta 4 was able to reduce effectively the plasma glucose concentration in streptozotocin-treated nude mice. Conophylline and betacellulin-delta 4 are effective in inducing differentiation of beta cells from progenitors.

    Web of Science

    researchmap

  • Gene therapy of liver tumors with human liver-specific nanoparticles (vol 14, pg 74, 2007)

    M. Ueda, Y. Iwasaki, T. Yamada, A. Kondo, M. Seno, K. Tanizawa, S. Kuroda, M. Sakamoto, M. Kitajima

    CANCER GENE THERAPY   14 ( 4 )   440 - 440   2007.4

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    DOI: 10.1038/sj.cgt.7701031

    Web of Science

    researchmap

  • AR42J細胞の分化に対する細胞外基質の影響

    濱本 耕平, 山田 聡子, 山本 頼綱, 小寺 力, 原 朱美, 妹尾 昌治, 小島 至

    糖尿病   50 ( Suppl.1 )   S - 349   2007.4

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

    researchmap

  • Characterization of bio-nanocapsule as a transfer vector targeting human hepatocyte carcinoma by disulfide linkage modification

    Tadahiro Nagaoka, Takayuki Fukuda, Shinnosuke Yoshida, Hirohito Nishimura, Dongwei Yu, Shun'ichi Kuroda, Katsuyuki Tanizawa, Akhko Kondo, Masakazu Ueda, Hidenori Yamada, Hiroko Tada, Masaharu Seno

    JOURNAL OF CONTROLLED RELEASE   118 ( 3 )   348 - 356   2007.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    The bio-nanocapsules (BNCs) composed of the recombinant envelope L-protein of hepatitis B virus constitute efficient delivery vectors specifically targeting human hepatocytes. Here, we have tried to enhance the stability of the BNCs because the L-proteins in the BNCs were aggregated due to random disulfide bridging when stored for a long period at 4 degrees C. The envelope protein contains fourteen cysteine residues in the S domain. Aggregation of the envelope proteins might be avoided if unessential cysteine residues are replaced or removed because the irreversible alkylation of the free sulfhydryl group protects against the aggregation and enhances the efficiency of encapsulation. In this study, the possibility of reducing the number of cysteine residues in the S domain to enhance the stability of the BNCs was assessed. The replacement of each cysteine residue by site-directed mutation showed that nine of fourteen cysteine residues were not essential to obtaining BNCs secreted into the culture media. Furthermore, upon evaluating the combination of these mutations, it was found that eight residues of replacement were acceptable. The mutant BNCs with replaced eight cysteine residues were not only more resistant against trypsin, but also more effective in transducing genes into human hepatoma-derived HepG2 cells than the original type BNC. Thus, we demonstrated that the minimized number of cysteine residues in the S domain could enhance the stability of the BNCs. (c) 2006 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jconrel.2006.12.020

    Web of Science

    researchmap

  • 脳腫瘍を標的としたバイオナノカプセルの開発

    筒井 佑美, 富澤 一仁, 西木 禎一, 大守 伊織, 名木田 真奈, 妹尾 昌治, 松井 秀樹

    日本生理学雑誌   69 ( 3 )   125 - 125   2007.3

     More details

    Language:Japanese   Publisher:(一社)日本生理学会  

    researchmap

  • Characteristics of hollow microtubes consisting of amorphous iron oxide nanoparticles produced by iron oxidizing bacteria, Leptothrix ochracea

    H. Hashimoto, S. Yokoyama, H. Asaoka, Y. Kusano, Y. Ikeda, M. Seno, J. Takada, T. Fujii, M. Nakanishi, R. Murakami

    JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS   310 ( 2 )   2405 - 2407   2007.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Some features of characteristic iron oxide sheaths which the iron oxidizing bacteria Leptothrix ochracea ( L. oceracea) formed were studied in order to make clear their morphology microstructure, chemical composition, and crystal structure through scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDX), and X-ray diffraction (XRD). Each sheath was a hollow tube with average outer and inner diameters of 1.1 and 1.4 m, respectively. Their length ranged from 10 to 200 mu m and the aspect ratio was 10-200. Each sheath was constructed by very small particles with a diameter of less than 100 nm. The hollow sheaths were mainly composed of Fe and O with small amounts of Si and P. The chemical composition analyzed by EDX was roughly Fe: Si: P = 80: 15: 5 with the exception of O. XRD measurement revealed that crystal structures of the sheath were similar to that of 2-line ferrihydrite. The sheath showed spin-glass-like magnetic properties. (c) 2006 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jmmm.2006.10.793

    Web of Science

    researchmap

  • Gene therapy of liver tumors with human liver-specific nanoparticles

    Y. Iwasaki, M. Ueda, T. Yamada, A. Kondo, M. Seno, K. Tanizawa, S. Kuroda, M. Sakamoto, M. Kitajima

    CANCER GENE THERAPY   14 ( 1 )   74 - 81   2007.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    The development of safe and efficient liver-specific gene delivery approaches offers new perspectives for the treatment of liver disease, in particular, liver cancer. We evaluated the therapeutic potential of hepatotropic nanoparticles for gene therapy of liver tumor. These nanoparticles do not contain a viral genome and display the hepatitis B virus L antigen, which is essential to confer hepatic specificity. It has not been shown whether a therapeutic effect could be obtained using L nanoparticles in a human liver tumor xenograft model. Rats bearing human hepatic ( NuE) and non-hepatic tumors were injected with L nanoparticles containing a green fluorescent protein ( GFP) expression plasmid. GFP expression was observed only in NuE-derived tumors but not in the non-hepatic tumor. The potential for treatment of liver tumors was analyzed using L nanoparticles containing the herpes simplex virus thymidine kinase gene, in conjunction with ganciclovir pro-drug administration. The growth of NuE-derived tumors in L particle-injected rats was significantly suppressed, but not of the non-hepatic tumor control. In summary, this is the first demonstration that nanoparticles could be used for delivery of therapeutic genes with anti-tumor activity into human liver tumors. This intravenous delivery system may be one of the major advantages as compared to many other viral vector systems.

    DOI: 10.1038/sj.cgt.7700990

    Web of Science

    researchmap

  • Conophylline and betacellulin delta 4 promote differentiation of pancreatic cells both in vitro and in vivo

    Tsutomu Kodera, Takeki Ogata, Yoritsuna Yamamoto, Kazuo Umezawa, Masaharu Seno, Yuji Tanaka, Itaru Kojima, Yasuhiro Watanabe

    JOURNAL OF PHARMACOLOGICAL SCIENCES   103   80P - 80P   2007

     More details

    Language:English   Publisher:JAPANESE PHARMACOLOGICAL SOC  

    Web of Science

    researchmap

  • バイオナノキャリアの開発とがん遺伝子治療への応用

    近藤昭彦, 黒田俊一, 谷澤克行, 妹尾昌治, 上田政和

    バイオテクノロジージャーナル   7 ( 1 )   41 - 47   2007

     More details

  • 心筋症治療薬の開発シーズ:好酸球由来塩基性タンパク質

    妹尾昌治

    ちゅうごく産業創造センター会報   No.74, p40-41.   2007

     More details

  • Anti-tumor effect in an in vivo model by human-derived pancreatic RNase with basic fibroblast growth factor insertional fusion protein through antiangiogenic properties

    Hiroshi Yagi, Masakazu Ueda, Hiromitsu Jinno, Koichi Aiura, Shuji Mikami, Hiroko Tada, Masaharu Seno, Hidenori Yamada, Masaki Kitajima

    CANCER SCIENCE   97 ( 12 )   1315 - 1320   2006.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BLACKWELL PUBLISHING  

    It is thought that the export of angiogenic fibroblast growth factors (FGF) from tumors may be involved in the onset of tumor angiogenesis. To create a new active targeting drug that inhibits the tumor angiogenic process without toxicities to normal cells, human basic FGF (h-bFGF) was inserted genetically into the Gly89 position of cross-linked RNase1 (the ribonuclease inhibitor protein [RI] binding site of cross-linked human pancreatic RNase) to prevent stereospecific binding to RI. The resultant insertional-fusion protein (CL-RFN89) was active both as h-bFGF and as RNase1. Furthermore, it acquired an additional ability of evading RI through steric blockade of RI binding caused by the fused h-bFGF domain. In the present study, the effect of the resultant protein, CL-RFN89, on the antitumor response though its antiangiogenic properties was investigated in an in vivo model. Continuous systemic treatment with CL-RFN89 significantly inhibited the growth of human A431 squamous cell carcinomas in vivo. Seven days of treatment with CL-RFN89 resulted in a 58.2% inhibition of tumor growth compared with control mice (P &lt; 0.0001). Furthermore, immunohistochemistry using a rat antimouse CD31 antibody showed that treatment with CL-RFN89 reduced tumor vascularization. These findings identify CL-RFN89 as a potent systemic inhibitor of tumor growth as a result of its antiangiogenic properties. This protein appears to be a new systemic antitumor agent.

    DOI: 10.1111/j.1349-7006.2006.00336.x

    Web of Science

    researchmap

  • Secretory production system of bionanocapsules using a stably transfected insect cell line

    Takuya Shishido, Masaru Muraoka, Masakazu Ueda, Masaharu Seno, Katsuyuki Tanizawa, Shun'ichi Kuroda, Hideki Fukuda, Akihiko Kondo

    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY   73 ( 3 )   505 - 511   2006.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER  

    Bionanocapsules (BNCs) are hollow nanoscale particles composed of L protein of the hepatitis B virus surface antigen that represent specific affinity for human hepatocytes. BNCs can transfer genes and drugs into human hepatocytes efficiently and specifically. BNC can be expressed in yeast cells. In this study, we developed a new L particle production system using a stably transfected insect cell line. For this purpose, we established a host vector system using the Trichoplusia ni insect cell line. L particles were efficiently secreted by the overexpression of the L protein, which was fused to the secretion signal peptide. The concentration of L particles was reached approximately 1.7 mu g/ml in 5 days during cultivation in a serum-free medium without antibiotic selective pressure. The production of L particles was maintained for at least 75 days. The secretory production of L particles facilitated their easy purification by chromatography. Furthermore, it was demonstrated that purified L particles can transfect only human hepatocytes. Therefore, an insect cell expression system is an attractive tool for the production of BNC.

    DOI: 10.1007/s00253-006-0486-3

    Web of Science

    researchmap

  • Hydrogen peroxide and endothelin-1 are novel activators of betacellulin ectodomain shedding

    Michael P. Sanderson, Catherine A. Abbott, Hiroko Tada, Masaharu Seno, Peter J. Dempsey, Andrew J. Dunbar

    JOURNAL OF CELLULAR BIOCHEMISTRY   99 ( 2 )   609 - 623   2006.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-LISS  

    The betacellulin precursor (pro-BTC) is a novel substrate for ADAM10-mediated ectodomain shedding. In this report, we investigated the ability of novel physiologically relevant stimuli, including G-protein coupled receptor (GPCR) agonists and reactive oxygen species (ROS), to stimulate pro-BTC shedding. We found that in breast adenocarcinoma MCF7 cells overexpressing pro-BTC, hydrogen peroxide (H2O2) was a powerful stimulator of ectodomain shedding. The stimulation of pro-BTC shedding by H2O2 was blocked by the broad-spectrum metalloprotease inhibitor TAPI-0 but was still functional in ADAM17 (TACE)-deficient stomach epithelial cells indicating the involvement of a distinct metalloprotease. H2O2-induced pro-BTC shedding was blocked by co-culturing cells in the anti-oxidant N-acetyl-L-cysteine but was unaffected by culture in calcium-deficient media. By contrast, calcium ionophore, which is a previously characterized activator of pro-BTC shedding, was sensitive to calcium depletion but was unaffected by co-culture with the anti-oxidant, identifying a clear distinction between these stimuli. We found that in vascular smooth muscle cells overexpressing pro-BTC, the GPCR agonist endothelin-1 (ET-1) was a strong inducer of ectodomain shedding. This was blocked by a metalloprotease inhibitor and by overexpression of catalytically inactive E385AADAM10. However, overexpression of wild-type ADAM10 or ADAM17 led to an increase in ET-1-induced pro-BTC shedding providing evidence for an involvement of both enzymes in this process. This study identifies ROS and ET-1 as two novel inducers of pro-BTC shedding and lends support to the notion of activated shedding occurring under the control of physiologically relevant stimuli. J. Cell. Biochem. 99: 609-623, 2006. (c) 2006 Wiley-Liss, Inc.

    DOI: 10.1002/jcb.20968

    Web of Science

    researchmap

  • FGF挿入型のヒト組み替えbFGF-RNaseを用いた腫瘍増殖抑制効果の検討

    八木 洋, 上田 政和, 神野 浩光, 相浦 浩一, 妹尾 昌治, 多田 宏子, 山田 秀徳, 北島 政樹

    日本癌治療学会誌   41 ( 2 )   697 - 697   2006.9

     More details

    Language:Japanese   Publisher:(一社)日本癌治療学会  

    researchmap

  • FGF挿入型のヒト組み替えbFGF-RNaseを用いた腫瘍増殖抑制効果の検討

    八木 洋, 上田 政和, 神野 浩光, 相浦 浩一, 妹尾 昌治, 多田 宏子, 山田 秀徳, 北島 政樹

    外科と代謝・栄養   40 ( 3 )   132 - 132   2006.6

     More details

    Language:Japanese   Publisher:日本外科代謝栄養学会  

    researchmap

  • Denatured and reversibly cationized p53 readily enters cells and simultaneously folds to the functional protein in the cells

    H Murata, M Sakaguchi, J Futami, M Kitazoe, T Maeda, H Doura, M Kosaka, H Tada, M Seno, N Huh, H Yamada

    BIOCHEMISTRY   45 ( 19 )   6124 - 6132   2006.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    Cationization is a powerful strategy for internalizing a protein into living cells. On the other hand, a reversibly cationized denatured protein through disulfide bonds is not only soluble in water but also able to fold to the native conformation in vitro. When these advantages in cationization were combined, we developed a novel method to deliver a denatured protein into cells and simultaneously let it fold to express its function within cells. This "in-cell folding" method enhances the utility of recombinant proteins expressed in Escherichia coli as inclusion bodies; that is, the recombinant proteins in inclusion bodies are solubilized by reversible cationization through cysteine residues by disulfide bonds with aminopropyl methanethiosulfonate or pyridyldithiopropionylpolyethylenimine and then incubated with cells without an in vitro folding procedure. As a model protein, we investigated human tumor-suppressor p53. Treatment of p53-null Saos-2 cells with reversibly cationized p53 revealed that all events examined as indications of the activation of p53 in cells, such as reduction of disulfide bonds followed by tetramer formation, localization into the nucleus, induction of p53 target genes, and induction of apoptosis of cells, occurred. These results suggest that reversible cationization of a denatured protein through cysteine residues is an alternative method for delivery of a functional protein into cells. This method would be very useful when a native folded protein is not readily available.

    DOI: 10.1021/bi052642a

    Web of Science

    researchmap

  • Mechanisms of the growth-inhibitory effect of the RNase-EGF fused protein against EGFR-overexpressing cells

    S Hoshimoto, M Ueda, H Jinno, M Kitajima, J Futami, M Seno

    ANTICANCER RESEARCH   26 ( 2A )   857 - 863   2006.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT INST ANTICANCER RESEARCH  

    Background: We previously showed the usefulness of a fused protein of human pancreatic ribonuclease1 (hRNase1) with human epidermal growth factor (hEGF) for molecular targeting of EGF receptor (EGFR)-overexpressing cells. In this study, the mechanisms underlying the inhibition of cell growth by RNase-EGF fused proteins was confirmed. Materials and Methods: Des. 1-7 hRNase1 was genetically fused to hEGF. The fused proteins were expressed and isolated from Escherichia coli. The internalization of hRNase1-hEGF was confirmed by confocal fluorescence microscopy. The growth-inhibitory effect of the fused proteins was evaluated by MTT assay. Results: FITC-labelled hRNase1-hEGF was internalized into EGFR-overexpressing A431 cells. The internalization was not observed in A 431 cells pre-treated with hEGF and EGFR-deficient H69 cells. The growth-inhibitory effect of des.1-7 hRNase1-hEGF against A431 cells was statistically significantly more pronounced than that of hRNase1-hEGF. Conclusion: RNase-EGF fused proteins are internalized through EGFR and inhibit cell growth by exerting their ribonucleolytic activity in the cytosol.

    Web of Science

    researchmap

  • Engineered bio-nanocapsules, the selective vector for drug delivery system

    DW Yu, T Fukuda, Tuoya, S Kuroda, K Tanizawa, A Kondo, M Ueda, T Yamada, H Tada, M Seno

    IUBMB LIFE   58 ( 1 )   1 - 6   2006.1

     More details

    Language:English   Publisher:TAYLOR & FRANCIS INC  

    The bio-nanocapsule (BNC) is our concept of artificial hollow nanoparticles that have been designed and produced through biotechnological procedures. We proposed an empty virus-like particle, which consists of a recombinant L envelope protein of hepatitis B virus (HBV) and a lipid derived from the host cell, as an engineered BNC. Although this BNC was first developed as an immunogen of hepatitis B vaccine, the pre-S1 region in N-terminus of L envelope protein confers hepatocyte specific infectivity of HBV on the BNC. This recombinant BNC is now being developed as a novel platform of drug delivery system (DDS) vector for selective delivery.

    DOI: 10.1080/15216540500484368

    Web of Science

    researchmap

  • Separation of Dual Affinity of Betacellulin to ErbB Receptors

    T. Nagaoka, H. Tada, H. Yamada, M. Seno

    MOLECULAR BIOLOGY OF THE CELL   17   2006

     More details

    Language:English   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Development of dds using hollow bio-nanoparticles and their commercialization

    Akihiko Kondo, Shun-ichi Kuroda, Katsuyuki Tanizawa, Masaharu Seno, Masakazu Ueda

    Drug Delivery System   21 ( 4 )   435 - 443   2006

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    We succeeded overproduction of the HBV envelope L particles with an approximate average particle size of 80 nm in yeast cells. Because the L particle is an empty bionanoparticles containing no viral DNA, it can be used as a safe and efficient carrier for human liver-specific delivery (pinpoint delivery) of drug and gene. In addition, genetically engineered L particles that are able to target to various organs were constructed by deleting the hepatocyte binding domain of L protein (pre-S region) and displaying targeting peptide or protein ligands. Therefore, bionanoparticles are a novel nano-carrier applicable to the broad range of pinpoint DDS. © 2006, THE JAPAN SOCIETY OF DRUG DELIVERY SYSTEM. All rights reserved.

    DOI: 10.2745/dds.21.435

    Scopus

    researchmap

  • Betacellulin-delta 4, a novel differentiation factor for pancreatic beta-cells, ameliorates glucose intolerance in streptozotocin-treated rats

    T Ogata, AJ Dunbar, Y Yamamoto, Y Tanaka, M Seno, Kojima, I

    ENDOCRINOLOGY   146 ( 11 )   4673 - 4681   2005.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ENDOCRINE SOC  

    We previously described a novel alternatively spliced mRNA transcript of the betacellulin (BTC) gene. This splice isoform, termed BTC-delta 4, lacks the C-loop of the epidermal growth factor motif and the transmembrane domain as a result of exon 4 'skipping'. In this study, we expressed BTC-delta 4 recombinantly to explore its biological function. When BTC-delta 4 was expressed in COS-7 cells, it was secreted largely into the culture medium, in contrast to BTC. Unlike BTC, highly purified recombinant BTC-delta 4 produced in Escherichia coli failed to bind or induce tyrosine phosphorylation of either ErbB1 or ErbB4, nor did it antagonize the binding of BTC to these receptors. Consistent with this, BTC-delta 4 failed to stimulate DNA synthesis in Balb/c 3T3 and INS-1 cells. However, BTC-delta 4 induced differentiation of pancreatic beta-cells; BTC-delta 4 converted AR42J cells to insulin-producing cells. When recombinant BTC-delta 4 was administered to streptozotocin-treated neonatal rats, it reduced the plasma glucose concentration and improved glucose tolerance. Importantly, BTC-delta 4 significantly increased the insulin content, the beta-cell mass, and the numbers of islet-like cell clusters and PDX-1-positive ductal cells. Thus, BTC-delta 4 is a secreted protein that stimulates differentiation of beta-cells in vitro and in vivo in an apparent ErbB1- and ErbB4- independent manner. The mechanism by which BTC-delta 4 exerts this action on beta-cells remains to be defined but presumably involves an, as yet, unidentified unique receptor.

    DOI: 10.1210/en.2005-0456

    Web of Science

    researchmap

  • Identification of cell surface marker candidates on SV-T2 cells using DNA microarray on DLC-coated glass

    Tuoya, K Hirayama, T Nagaoka, DW Yu, T Fukuda, H Tada, H Yamada, M Seno

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   334 ( 1 )   263 - 268   2005.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We analyzed gene expression profiles of normal mouse fibroblast BALB/c 3T3 cells and its SV40 transformant SV-T2 cells using our originally developed cell surface marker DNA microarray, which is prepared on a diamond-like carbon-coated glass. As a result, CD62L and IL-6 receptor alpha gene expressions were upregulated in SV-T2 and were thought to be candidates for cell surface markers of the cells. The result of microarray analysis was validated by real-time quantitative PCR, immunohistochemistry and biological assays. These data show that our cell surface marker DNA microarray should be useful in finding the candidates of cell type-specific surface markers. (c) 2005 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2005.06.083

    Web of Science

    researchmap

  • 細胞および組織特異的遺伝子導入を可能にするバイオナノカプセル

    YAMADA Tadanori, SENO Masaharu, UEDA Masakazu, KONDO Akihiko, TANIZAWA Katsuyuki, KURODA Shun'ichi

    生物工学会誌   83・8・380-383   2005.8

     More details

    Language:Japanese  

    researchmap

  • Anti-angiogenic effect of an insertional fusion protein of human basic fibroblast growth factor and ribonuclease-1

    T Hayashida, M Ueda, K Aiura, H Tada, M Onizuka, M Seno, H Yamada, M Kitajima

    PROTEIN ENGINEERING DESIGN & SELECTION   18 ( 7 )   321 - 327   2005.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OXFORD UNIV PRESS  

    Human pancreatic ribonuclease-1 (RNase1) does not exhibit its cytotoxicity unless it is artificially internalized into the cytosol. Furthermore, once it encounters the cytosolic RNase inhibitor (RI), the activity of RNase1 is seriously reduced. To achieve the cellular targeting of RNase1 and the blocking of RI binding simultaneously, the basic fibroblast growth factor (bFGF) sequence was inserted into RNase1 at the RI binding site using a gene fusion technique. The effect of this fusion protein, CL-RFN89, on the angiogenesis, which was accelerated by FGF-FGF receptor interaction, was investigated. It was shown by using fluorescein-labeled CL-RFN89, that the binding to human umbilical vein endothelial cells (HUVECs) was dependent on the existence of the FGF receptors. In addition, CL-RFN89 inhibited the cellular growth of HUVECs in vitro and also inhibited the tube formation, using a three-dimensional tube formation assay. Furthermore, this fusion protein was shown to prevent in vivo tumor cell-induced angiogenesis, using the mouse dorsal air sac assay. These results demonstrated that CL-RFN89 inhibits angiogenesis in vitro and in vivo and that it can be expected to be a potent antiangiogenic agent.

    DOI: 10.1093/protein/gzi040

    Web of Science

    researchmap

  • The specific delivery of proteins to human liver cells by engineered bio-nanocapsules

    DW Yu, C Amano, T Fukuda, T Yamada, S Kuroda, K Tanizawa, A Kondo, M Ueda, H Yamada, H Tada, M Seno

    FEBS JOURNAL   272 ( 14 )   3651 - 3660   2005.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BLACKWELL PUBLISHING  

    A bio-nanocapsule (BNC), composed of the surface antigen (sAg) of the hepatitis B virus, is an efficient nanomachine with which to accomplish the liver-specific delivery of genes and drugs. Approximately 110 molecules of sAg are associated to form a BNC particle with an average diameter of 130 nm. The L protein is an sAg peptide composed mainly of preS and S regions. The preS region, with specific affinity for human hepatocytes, is localized in the N-terminus. The S region following the preS has two transmembrane regions responsible for the formation of particles. In this study, the fusion of emerald green fluorescent protein (EGFP) at the C-terminus of the S region was designed to deliver proteins to human hepatocytes. Truncation of the C-terminus of the S region was required to obtain sufficient expression levels in Cos7 cells. The nanoparticles that were produced delivered EGFP to human hepatoma cells, displaying the EGFP moiety outside, or enclosing it inside. However, only a single orientation characterizes the particle, so that either type of L fusion particle could be effectively and independently separated by an antibody affinity column. The dual C-terminal topologies of the L fusion particles designed in this study could be applied to various proteins for the C-terminal moiety of the L fusion proteins, depending on the character of the proteins, such as cytoplasmic proteins, as well as cytokines or ligands to cell surface receptors. We suggest that this fusion design is the most efficient way to prepare a BNC that delivers proteins to specific cells or tissues.

    DOI: 10.1111/j.1742-4658.2005.04790.x

    Web of Science

    researchmap

  • Protein transduction assisted by polyethylenimine-cationized carrier proteins

    M Kitazoe, H Murata, J Futami, T Maeda, M Sakaguchi, M Miyazaki, M Kosaka, H Tada, M Seno, N Huh, M Namba, M Nishikawa, Y Maeda, H Yamada

    JOURNAL OF BIOCHEMISTRY   137 ( 6 )   693 - 701   2005.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    Previously, we have reported that cationized-proteins covalently modified with polyethylenimine (PEI) (direct PEI-cationization) efficiently enter cells and function in the cytosol [Futami et al. (2005) J. Biosci. Bioeng. 99,95-1031. However, it may be more convenient if a protein could be delivered into cells just by mixing the protein with a PEI-cationized carrier protein having a specific affinity (indirect PEI-cationization). Thus, we prepared PEI-cationized avidin (PEI-avidin), streptavidin (PEI-streptavidin), and protein G (PEI-protein G), and examined whether they could deliver biotinylated proteins and antibodies into living cells. PEI-avidin (and/or PEI-streptavidin) carried biotinylated GFPs into various mammalian cells very efficiently. A GFP variant containing a nuclear localization signal was found to arrive even in the nucleus. The addition of a biotinylated RNase A derivative mixed with PEI-streptavidin to a culture medium of 3T3-SV-40 cells resulted in remarkable cell growth inhibition, suggesting that the biotinylated RNase A derivative entered cells and digested intracellular RNA molecules. Furthermore, the addition of a fluorescein-labeled antiS100C (beta-actin binding protein) antibody mixed with PEI-protein G to human fibroblasts resulted in the appearance of a fluorescence image of actin-like filamentous structures in the cells. These results indicate that indirect PEI-cationization using non-covalent interaction is as effective as the direct PEI-cationization for the transduction of proteins into living cells and for expression of their functions in the cytosol. Thus, PEI-cationized proteins having a specific affinity for certain molecules such as PEI-streptavidin, PEI-avidin and PEI-protein G are concluded to be widely applicable protein transduction carrier molecules.

    DOI: 10.1093/jb/mvi081

    Web of Science

    researchmap

  • Intracellular delivery of proteins into mammalian living cells by polyethylenimine-cationization

    Junichiro Futami, Midori Kitazoe, Takashi Maeda, Emiko Nukui, Masakiyo Sakaguchi, Jun Kosaka, Masahiro Miyazaki, Megumi Kosaka, Hiroko Tada, Masaharu Seno, Junzo Sasaki, Nam-Hu Huh, Masayoshi Namba, Hidenori Yamada

    Journal of Bioscience and Bioengineering   99 ( 2 )   95 - 103   2005

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Society of Fermentation and Bioengineering  

    In the post-genomic era, there is pressing need for development of protein manipulation methodology to analyze functions of proteins in living cells. For this purpose, techniques to deliver functional proteins into living cells are currently being evaluated as alternative approaches to the introduction of transcriptionally active DNA. Here, we describe a novel method for efficient protein transduction into living cells in which a protein is simply cationized with polyethylenimine (PEI) by limited chemical conjugation. PEI-cationized proteins appear to adhere to the cell surface by ionic charge interaction and then internalize into cells in a receptor- and transporter-independent fashion. Since PEI is an organic macromolecule with a high cationic-charge density, limited coupling with PEI results in endowment of sufficient cationic charge to proteins without causing serious decline in their fundamental functions. A number of PEI-cationized proteins, such as ribonuclease (RNase), green fluorescent protein (GFP) and immunoglobulin (IgG), efficiently entered cells and functioned in the cytosol. Our results suggest that protein cationization techniques using PEI will be useful for the development of protein transduction technology. © 2005, The Society for Biotechnology.

    DOI: 10.1263/jbb.99.95

    Scopus

    PubMed

    researchmap

  • 革新的なナノキャリア:中空バイオナノ粒子によるピンポイントDDS

    近藤昭彦, 黒田俊一, 谷澤克行, 妹尾昌治, 上田政和

    バイオインダストリー   22 ( 5 )   22 - 27   2005

     More details

  • Conophylline and betacellulin-delta 4: An effective combination to induce differentiation of pancreatic beta cells

    RI Kitamura, T Ogata, Y Tanaka, MH Seno, Takei, I, K Umezawa, Kojima, I

    DIABETES   54 ( 2 )   A393 - A393   2005

     More details

    Language:English   Publisher:AMER DIABETES ASSOC  

    DOI: 10.1507/endocrj.K06-199

    Web of Science

    researchmap

  • 中空バイオナノ粒子を用いたピンポイントDDSの開発

    近藤昭彦, 黒田俊一, 谷澤克行, 妹尾昌治

    Chemical Engineering   50 ( 5 )   351 - 356   2005

     More details

  • Differentiation of adult hepatic stem-like cells into pancreatic endocrine cells

    S Yamada, K Terada, Y Ueno, T Sugiyama, M Seno, Kojima, I

    CELL TRANSPLANTATION   14 ( 9 )   647 - 653   2005

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:COGNIZANT COMMUNICATION CORP  

    To apply cell transplantation for treatment of diabetes mellitus, a sufficient number of beta-cell sources are required. In the present study, we examined whether an epithelial cell line obtained from normal adult rat liver, namely hepatic stem-like (HSL) cells, which can be converted to both hepatocytes and billiary epithelial cells, could be a potential beta-cell source. The growth speed of HSL cells was rapid and these cells were easily expanded in vitro. Bipotential hepatic stem cells, HSL cells, also expressed PGP9.5, which is expressed in neurons, beta-cells, and progenitor cells of the pancreatic endocrine cells as well. Sodium butyrate induced morphological changes in HSL cells and converted them into flattened cells with large cytoplasm. When HSL cells were incubated with a combination of 5 mM sodium butyrate and I nM betacellulin, most of the cells were converted into morphologically neuron-like cells. RT-PCR analysis revealed that a series of transcriptional factors involved in differentiation of pancreatic endocrine cells was induced by the treatment with sodium butyrate and betacellulin. mRNAs for insulin, pancreatic polypeptide, and somatostatin were also observed. Immunoreactive pancreatic polypeptide, somatostatin, and insulin were detected in sodium butyrate and betacellulin-treated HSL cells. In conclusion, HSL cells obtained from adult normal liver also have the potential to differentiate into pancreatic endocrine cells in vitro. HSL cells may be one of the potential beta-cell sources for cell transplant therapy for insulin-dependent diabetes.

    DOI: 10.1016/j.ssc.2005.01.003

    Web of Science

    researchmap

  • Characterization of cell surface marker proteins in SV-T2 cells

    Y Tuo, K Hirayama, T Nagaoka, H Tada, H Yamada, M Seno

    MOLECULAR BIOLOGY OF THE CELL   15   324A - 325A   2004.11

     More details

    Language:English   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Epithelial mesenchymal transition is a characteristic of hyperplasias and tumors in mammary gland from MMTV-cripto-1 transgenic mice

    L Strizzi, C Bianco, N Normanno, M Seno, C Wechselberger, B Wallace-Jones, NI Khan, M Hirota, YP Sun, M Sanicola, DS Salomon

    JOURNAL OF CELLULAR PHYSIOLOGY   201 ( 2 )   266 - 276   2004.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-LISS  

    Epithelial-mesenchymal transition (EMT) facilitates migration and invasion of epithelial tumor cells. Cripto-1 (CR-1), a member of the epidermal growth factor-CFC protein family increases migration of cells in vitro. Here the expression of molecular markers and signaling molecules characteristic of EMT were assessed in mammary gland hyperplasias and tumors from mice expressing the human CR-1 transgene by the MMTV promoter (MMTV-CR-1) and in mouse mammary epithelial cell line HC-11 overexpressing CR-1 (HC-11/CR-1). Western blot analysis showed decreased expression of E-cadherin in MMTV-CR-1 tumors and in HC-11/CR-1 cells. The expression of N-cadherin, vimentin, cyclin-D1, and of the zinc-finger transcription factor, snail, was increased in MMTV-CR-1 tumors. Increased snail mRNA was also found in HC-11/CR-1 cells. Expression of phosphorylated (N-c-Src, P-focal adhesion kinase (FAK), P-Akt, P-glycogen synthease kinase 30 (GSK-3beta), dephosphorylated (DP)-beta-catenin, and various integrins such as, alpha 3, alpha v, beta 1, beta 3, and beta 4 was also increased in MMTV-CR-1 tumors. Immunohistochemistry showed positive staining for vimentin, N-cadherin, cyclin-D1, smooth muscle actin, fibronectin, snail, and beta-catenin in MMTV-CR-1 tumor sections. HC-11/CR-1 cells treated with the c-Src inhibitor PP2 reduced the expression of P-c-Src and of P-FAK, P-Akt, P-GSK-3beta, DP-beta-catenin all known to be activated by c-Src. Migration of HC-1I/CR-1 cells was also reduced by PP2 treatment. These results suggest that CR-1 may play a significant role in promoting the increased expression of markers and signaling molecules associated with EMT. (C) 2004 Wiley-Liss, Inc.

    DOI: 10.1002/jcp.20062

    Web of Science

    researchmap

  • Reversal of streptozotocin-incluced hyperglycemia by transplantation of pseudoislets consisting of beta cells derived from ductal cells

    T Ogata, KY Park, M Seno, Kojima, I

    ENDOCRINE JOURNAL   51 ( 3 )   381 - 386   2004.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN ENDOCRINE SOCIETY  

    The present study was conducted in an attempt to treat streptozotocin (STZ)-induced hyperglycemia by transplanting beta cells derived from pancreatic ductal cells. Ductal cells obtained from neonatal rats were cultured in vitro. Approximately 70% of the cells were converted to insulin-secreting cells by incubating with betacellulin and activin A. Differentiated cells responded to a depolarizing concentration of potassium, tolbutamide and a high concentration of glucose, and insulin secretion increased by 2.5-, 2.3- and 1.6-fotd, respectively. We then prepared pseudoislets using the differentiated cells, which exhibited greatly improved glucose-responsiveness, with a high concentration of glucose inducing a 3-fold increase in insulin secretion. We transplanted these pseudoislets into the portal vein of STZ-treated nude mice. Before transplantation, the plasma glucose concentration was above 400 mg/dl, and after transplantation it was markedly reduced, the effect of which persisted for two weeks. These results indicate that STZ-induced hyperglycemia can be treated by transplanting pseudoislets consisting of 0 cells derived from ductal cells.

    DOI: 10.1507/endocrj.51.381

    Web of Science

    researchmap

  • Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction

    H Tada, M Onizuka, K Muraki, W Masuzawa, J Futami, M Kosaka, M Seno, H Yamada

    FEBS LETTERS   568 ( 1-3 )   39 - 43   2004.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Basic fibroblast growth factor (bFGF) was inserted in the middle of human ribonuclease 1 (RNase1) sequence at an RNase inhibitor (RI)-binding site (Gly89) by a new gene fusion technique, insertional-fusion. The resultant insertional-fusion protein (CL-RFN89) was active both as bFGF and as RNase. Furthermore, it acquired an additional ability of evading RI through steric blockade of RI-binding caused by fused bFGF domain. As a result, CL-RFN89 showed stronger growth inhibition on B16/BL6 melanoma cells than an RI-sensitive tandem fusion protein. Thus, the insertional-fusion technique increases accessible positions for gene fusion on RNase, resulting in construction of a potent cytotoxic RNase. (C) 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.febslet.2004.05.007

    Web of Science

    researchmap

  • Activin A and betacellulin - Effect on regeneration of pancreatic beta-cells in neonatal streptozotocin-treated rats

    L Li, ZH Yi, M Seno, Kojima, I

    DIABETES   53 ( 3 )   608 - 615   2004.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER DIABETES ASSOC  

    Activin A and betacellulin (BTC) are thought to regulate differentiation of pancreatic beta-cells during development and regeneration of beta-cells in adults. In the present study, we used neonatal rats treated with streptozotocin (STZ) to investigate the effects of activin A and BTC on regeneration of pancreatic beta-cells. One-dayold Sprague-Dawley rats were injected with STZ (85 mug/g) and then administered for 7 days with activin A and/or BTC. Treatment with activin A and BTC significantly reduced the plasma glucose concentration and the plasma glucose response to intraperitoneal glucose loading. The pancreatic insulin content and beta-cell mass in rats treated with activin A and BTC were significantly increased compared with the control group on day 8 and at 2 months. Treatment with activin A and BTC significantly increased the DNA synthesis in preexisting beta-cells, ductal cells, and 8-cells. The number of islet cell-like clusters (ICCs) and islets was significantly increased by treatment with activin A and BTC. In addition, the number of insulin/somatostatin-positive cells and pancreatic duodenal homeobox-1/somatostatin-positive cells was significantly increased. These results indicate that, in neonatal STZ-treated rats, a combination of activin A and BTC promoted regeneration of pancreatic beta-cells and improved glucose metabollism. in adults.

    DOI: 10.2337/diabetes.53.3.608

    Web of Science

    researchmap

  • Novel tissue and cell type-specific gene/drug delivery system using surface engineered hepatitis B virus nano-particles

    Tadanori Yamada, Masakazu Ueda, Masaharu Seno, Akihiko Kondo, Katsuyuki Tanizawa, Shun'ichi Kuroda

    Current Drug Targets - Infectious Disorders   4 ( 2 )   163 - 167   2004

     More details

    Language:English  

    The hepatitis B virus (HBV) surface antigen (HBsAg) L particle is a hollow nano-scale particle. HBsAg L particles have many properties that make them useful for in vivo gene transfer vectors and drug delivery systems. Gene therapy so far has required the in vivo pinpoint delivery of genetic materials into the target organs and cells. Gene transfer by HBsAg L particles might be an attractive method, since their tropism is the same as that of HBV. The HBsAg L particles are able to deliver therapeutic payloads with high specificity to human hepatocytes. In addition, the specificity of L particle can be altered by displaying various cell-binding molecules on the surface. Our results indicate that the L particle is suitable for a cell- and tissue-specific gene/drug transfer vector. In this review, we discuss HBsAg L particles as a gene/drug transfer vector and its potential for the treatment of infectious diseases. © 2004 Bentham Science Publishers Ltd.

    DOI: 10.2174/1568005043341037

    Scopus

    PubMed

    researchmap

  • 中空バイオナノ粒子が拓く新しい医療技術

    黒田俊一, 山田忠範, 妹尾昌治, 近藤昭彦, 上田政和, 谷澤克行

    化学工業   vol.55, no.12, pp.936-942   2004

     More details

  • Pinpoint drug delivery system using hollow bio-nanoparticles Reviewed

    T Yamada, M Seno, A Kondo, M Ueda, K Tanizawa, S Kuroda

    KOBUNSHI RONBUNSHU   61 ( 12 )   606 - 612   2004

     More details

    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:SOC POLYMER SCIENCE JAPAN  

    Hepatitis B virus envelope L proteins produced in yeast cells form hollow nanoparticles (L particles, average diameter 220 nm) displaying human liver-specific receptor. Recently, the L particles were found to incorporate genes, proteins, and drugs, and act as an efficient pinpoint delivery system to human liver-derived tissues in xenograft models. By substituting the epidermal growth factor (EGF) for human liver-specific receptor, the mutated L particles showed the affinity to the EGF receptor, not to human liver. Other similar HBV envelope proteins, e.g., M and S particles, have already been commercialized for hepatitis B vaccine, strongly suggesting the safety of L particles in human. These results indicate that the hollow bio-nanoparticles are a promising candidate for the next-generation platform of DDS, especially that related to gene therapy.

    Web of Science

    researchmap

  • Betacellulin improves glucose metabolism by promoting conversion of intraislet precursor cells to beta-cells in streptozotocin-treated mice

    L Li, M Seno, H Yamada, Kojima, I

    AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM   285 ( 3 )   E577 - E583   2003.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER PHYSIOLOGICAL SOC  

    Beta-cellulin (BTC) induces differentiation of pancreatic beta-cells and promotes regeneration of beta-cells in experimental diabetes. The present study was conducted to determine if BTC improved glucose metabolism in severe diabetes induced by a high dose of streptozotocin (STZ) in mice. Male ICR mice were injected with 200 mug/g ip STZ, and various doses of BTC were administered daily for 14 days. The plasma glucose concentration increased to a level of &gt;500 mg/dl in STZ-injected mice. BTC (0.2 mug/g) significantly reduced the plasma glucose concentration, but a higher concentration was ineffective. The effect of BTC was marked by day 4 but became smaller on day 6 or later. The plasma insulin concentration and the insulin content were significantly higher in mice treated with 0.1 and 0.2 mug/g BTC. BTC treatment significantly increased the number of beta-cells in each islet as well as the number of insulin-positive islets. Within islets, the numbers of 5-bromo-2-deoxyuridine/somatostatin-positive cells and pancreatic duodenal homeobox-1/somatostatin-positive cells were significantly increased by BTC. These results indicate that BTC improved hyperglycemia induced by a high dose of STZ by promoting neoformation of beta-cells, mainly from somatostatin-positive islet cells.

    DOI: 10.1152/ajpendo.00120.2003

    Web of Science

    researchmap

  • Nanoparticles for the delivery of genes and drugs to human hepatocytes

    T Yamada, Y Iwasaki, H Tada, H Iwabuki, MKL Chuah, T VandenDriessche, H Fukuda, A Kondo, M Ueda, M Seno, K Tanizawa, S Kuroda

    NATURE BIOTECHNOLOGY   21 ( 8 )   885 - 890   2003.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    Hepatitis B virus envelope L particles form hollow nanoparticles displaying a peptide that is indispensable for liver-specific infection by hepatitis B virus in humans. Here we demonstrate the use of L particles for the efficient and specific transfer of a gene or drug into human hepatocytes both in culture and in a mouse xenograft model. In this model, intravenous injection of L particles carrying the gene for green fluorescent protein (GFP) or a fluorescent dye resulted in observable fluorescence only in human hepatocellular carcinomas but not in other human carcinomas or in mouse tissues. When the gene encoding human clotting factor IX was transferred into the xenograft model using L particles, factor IX was produced at levels relevant to the treatment of hemophilia B. The yeast-derived L particle is free of viral genomes, highly specific to human liver cells and able to accommodate drugs as well as genes. These advantages should facilitate targeted delivery of genes and drugs to the human liver.

    DOI: 10.1038/nbt843

    Web of Science

    researchmap

  • US venture capital for biotechnology Reviewed

    MD Dibner, M Trull, M Howell

    NATURE BIOTECHNOLOGY   21 ( 6 )   613 - 617   2003.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    DOI: 10.1038/nbt0603-613

    Web of Science

    researchmap

  • The cytotoxicity of a conjugate composed of human epidermal growth factor and eosinophil cationic protein

    H Jinno, M Ueda, S Ozawa, T Ikeda, M Kitajima, T Maeda, M Seno

    ANTICANCER RESEARCH   22 ( 6C )   4141 - 4145   2002.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT INST ANTICANCER RESEARCH  

    Background: Conventional targeted therapy with foreign proteins is highly immunogenic. In this study, we developed targeted therapy composed of human endogenous proteins and evaluated its efficacy in vitro. Materials and Methods: Human epidermal growth factor (EGF) was chemically linked to human eosinophil cationic protein (ECP). The cytotoxicity of the EGF-ECP conjugate was evaluated by MTT assay. Results: The conjugate showed dose-dependent cytotoxicity on EGF receptor (EGFR)-overexpressing BT-20 cells with an IC50 of 1.5 x 10(-7) M, whereas the IC50 of ECP alone was almost 10(-4) M, The conjugate had no detectable cytotoxicity against EGF receptor- deficient H69 cells. Excess EGF protected BT-20 cells from the cytotoxicity of the conjugate. Comparing the cytotoxicity and the level of EGFR expression, the cytotoxicity of the conjugate was positively correlated with the level of EGFR expression of each cell line. Conclusion: Conjugates composed solely of human proteins might be useful with less immunogenicity and less toxicity than the conventional immunotoxins for targeted therapy.

    Web of Science

    researchmap

  • RNase 3 (ECP) is an extraordinarily stable protein among human pancreatic-type RNases

    T Maeda, K Mahara, M Kitazoe, J Futami, A Takidani, M Kosaka, H Tada, M Seno, H Yamada

    JOURNAL OF BIOCHEMISTRY   132 ( 5 )   737 - 742   2002.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    There have been some attempts to develop immunotoxins utilizing human RNase as a cytotoxic domain of antitumor agents. We have recently shown that only human RNase 3 (eosinophil cationic protein, ECP) among five human pancreatic-type RNases excels in binding to the cell surface and has a growth inhibition effect on several cancer cell lines, even though the RNase activity of RNase 3 is completely inhibited by the ubiquitously expressed cytosolic RNase inhibitor. This phenomenon may be explained by that RNase 3 is very stable against proteolytic degradation because RNase 3 internalized through endocytosis could have a longer life time in the cytosol, resulting in the accumulation of enough of it to exceed the concentration of RNase inhibitor, which allows the degradation of cytosolic RNA molecules. Thus, we compared the stabilities of human pancreatic-type RNases (RNases 1-5) and bovine RNase A by means of guanidium chloride-induced denaturation experiments based on the assumption of a two-state transition for unfolding. It was demonstrated that RNase 3 is extraordinarily stabler than either RNase A or the other human RNases (by more than 25 kJ/mol). Thus, our data suggest that in addition to its specific affinity for certain cancer cell lines, the stability of RNase 3 contributes to its unique cytotoxic effect and that it is important to stabilize a human RNase moiety through protein engineering for the design of human RNase-based immunotoxins.

    Web of Science

    researchmap

  • 上皮増殖因子を用いたターゲッテイング療法の検討

    神野 浩光, 上田 政和, 池田 正, 北島 政樹, 妹尾 昌治

    日本癌治療学会誌   37 ( 2 )   373 - 373   2002.9

     More details

    Language:Japanese   Publisher:(一社)日本癌治療学会  

    researchmap

  • Optimum modification for the highest cytotoxicity of cationized ribonuclease

    J Futami, E Nukui, T Maeda, M Kosaka, H Tada, M Seno, H Yamada

    JOURNAL OF BIOCHEMISTRY   132 ( 2 )   223 - 228   2002.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    Cationization of a protein is considered to be a powerful strategy for internalizing a functional protein into cells. Cationized proteins appear to adsorb to the cell surface by electrostatic interactions, then enter the cell in a receptor- and transporter-independent fashion. Thus, in principle, all cell types appear to take up cationized proteins. Since ribonucleases (RNases) have a latent cytotoxic potential, cationized RNases could be useful cancer chemotherapeutics. In this study, we investigated the effect of the degree of cationization on the cytotoxicity of RNase A by modifying carboxyl groups with ethylenediamine. We found that there is an optimum degree of modification for cytotoxicity, in which 5 to 7 out of 11 carboxyl groups in RNase A are modified, toward MCF-7 and 3T3-SV-40 cells. More interestingly, the cytotoxicity of cationized RNase As correlates well with the value of [RNase activity] x [estimated concentration of RNase free from RNase inhibitor], mimicking the practical enzymatic activity of cationized RNase As in cytosol. The results indicate that cationization of a protein to an optimum level is important for maintaining protein function in the cytosol. Sophisticated protein cationization techniques will help to advance protein transduction technology.

    Web of Science

    researchmap

  • Solution structure of betacellulin, a new member of EGF-family ligands

    K Miura, H Doura, T Aizawa, H Tada, M Seno, H Yamada, K Kawano

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   294 ( 5 )   1040 - 1046   2002.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The solution structure of the EGF-like domain of betacellulin (BTCe), a newly discovered member of the epidermal growth factor (EGF) family, has been determined using two-dimensional nuclear magnetic resonance spectroscopy. This is the first report to identify the solution structure of the EGF-family ligand monomers that interact with both ErbB-1 and ErbB-4. The solution structure of BTCe was calculated using 538 NMR-derived restraints. The overall structure of BTCe was stabilized by three disulfide bonds, a hydrophobic core, and 23 hydrogen bonds. It appears that BTCe is comprised of five beta-strands and one short 3(10) helical turn. The secondary structural elements of BTCe are basically similar to those of the other EGF-family proteins, except that several significant variations of the structural properties were found. It is suggested that the structural variations between BTCe and the other EGF-family ligands may affect the specific receptor-recognition properties of EGF-family ligands. (C) 2002 Elsevier Science (USA). All rights reserved.

    DOI: 10.1016/S0006-291X(02)00585-5

    Web of Science

    researchmap

  • Combined expression of pancreatic duodenal homeobox 1 and islet factor 1 induces immature enterocytes to produce insulin

    H Kojima, T Nakamura, Y Fujita, A Kishi, M Fujimiya, S Yamada, M Kudo, Y Nishio, H Maegawa, M Haneda, H Yasuda, Kojima, I, M Seno, NCW Wong, R Kikkawa, A Kashiwagi

    DIABETES   51 ( 5 )   1398 - 1408   2002.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER DIABETES ASSOC  

    Immature rat intestinal stem cells (IEC-6) given the ability to express the transcription factor, pancreatic duodenal homeobox 1 (Pdx-1), yielded YK cells. Although these cells produced multiple enteroendocrine hormones, they did not produce insulin. Exposure of YK cells to 2 nmol/l betacellulin yielded BYK cells that showed the presence of insulin expression in cytoplasm and that secreted insulin into culture media. By examining the mechanism of differentiation in BYK cells, we found that another transcription factor, islet factor 1 (Isl-1) was newly expressed with the disappearance of Pax-6 expression in those cells after exposure to betacellulin. These results indicated that combined expression of Pdx-1 and Isl-1 in IEC-6 cells was required for the production of insulin. In fact, overexpression of both Pdx-1 and Isl-1 in IEC-6 cells (Isl-YK-12, -14, and -15 cells) gave them the ability to express insulin without exposure to betacellulin. Furthermore, implantation of the Isl-YK-14 cells into diabetic rats reduced the animals' plasma glucose levels; glucose levels dropped from 19.4 to 16.9 mmol/l 1 day after the injection of cells. As expected, the plasma insulin concentrations were 2.7 times higher in the diabetic rats injected with Isl-YK-14 cells compared to in controls. In summary, our results indicated that immature intestinal stem cells can differentiate into insulin-producing cells given the ability to express the transcription factors Pdx-1 and Isl-1.

    Web of Science

    researchmap

  • Cripto-1 activates nodal- and ALK4-dependent and -independent signaling pathways in mammary epithelial cells

    C Bianco, HB Adkins, C Wechselberger, M Seno, N Normanno, A De Luca, YP Sun, N Khan, N Kenney, A Ebert, KP Williams, M Sanicola, DS Salomon

    MOLECULAR AND CELLULAR BIOLOGY   22 ( 8 )   2586 - 2597   2002.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC MICROBIOLOGY  

    Cripto-1 (CR-1), an epidermal growth factor-CFC (EGF-CFC) family member, has a demonstrated role in embryogenesis and mammary gland development and is overexpressed in several human tumors. Recently, EGF-CFC proteins were implicated as essential signaling cofactors for Nodal, a transforming growth factor beta family member whose expression has previously been defined as embryo specific. To identify a receptor for CR-1, a human brain cDNA phage display library was screened using CR-1 protein as bait. Phage inserts with identity to ALK4, a type I serine/threonine kinase receptor for Activin, were identified. CR-1 binds to cell surface ALK4 expressed on mammalian epithelial cells in fluorescence-activated cell sorter analysis, as well as by coimmunoprecipitation. Nodal is coexpressed with mouse Cr-1 in the mammary gland, and CR-1 can phosphorylate the transcription factor Smad-2 in EpH-4 mammary epithelial cells only in the presence of Nodal and ALK4. In contrast, CR-1 stimulation of mitogen-activated protein kinase and AKT in these cells is independent of Nodal and ALK4, suggesting that CR-1 may modulate different signaling pathways to mediate its different functional roles.

    DOI: 10.1128/MCB.22.8.2586-2597.2002

    Web of Science

    researchmap

  • EGFRを分子標的としたRNase-EGF recombinantタンパクの殺細胞効果

    星本 相淳, 上田 政和, 神野 浩光, 相浦 浩一, 渡辺 靖夫, 北島 政樹, 二見 淳一郎, 妹尾 昌治

    日本外科学会雑誌   103 ( 臨増 )   532 - 532   2002.3

     More details

    Language:Japanese   Publisher:(一社)日本外科学会  

    researchmap

  • Detection and localization of Cripto-1 binding in mouse mammary epithelial cells and in the mouse mammary gland using an immunoglobulin-Cripto-1 fusion protein

    C Bianco, N Normanno, A De Luca, MR Maiello, C Wechselberger, Y Sun, N Khan, H Adkins, M Sanicola, B Vonderhaar, B Cohen, M Seno, D Salomon

    JOURNAL OF CELLULAR PHYSIOLOGY   190 ( 1 )   74 - 82   2002.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-LISS  

    Human Cripto-1 (CR-1), a member of the epidermal growth factor-CFC (EGF-CFC) family of peptides, is expressed in the developing mouse mammary gland and can modulate mammary epithelial cell migration, branching morphogenesis and milk protein expression in vitro. In order to screen for a CR-1 receptor and to identify potential CR-1 target tissues, we constructed a fusion protein comprising the EGF-like domain of CR-1 and the Fc domain of a human IgG1. The recombinant CR-1 fusion protein (CR-1-Fc) was biologically active as it was able to activate the ras/raf/mitogen activated protein kinase (MAPK) pathway and to inhibit transcription of the milk protein P-casein in NMuMG and HC-11 mouse mammary epithelial cells. By using immunocytochemistry and by an in situ enzyme-linked immunosorbent assay (ELISA), CR-1-Fc was found to specifically bind to NMuMG and HC-11 cells. Finally, immunohistochemical analysis using CR-1-Fc showed a specific localization of CR-1 binding to tissue sections from mouse mammary gland. In particular, more than 60% of the epithelial cells were intensely stained with the CR-1-Fc fusion protein in the lactating mouse mammary gland, whereas approximately 25% of the mammary epithelial cells were stained in the gland from pregnant mouse. Since expression of mouse cripto-1 (Cr-1) in the pregnant and lactating mouse mammary gland as well as its presence in milk has been previously demonstrated, these data strongly suggest that an autocrine pathway involving Cr-1 and its putative receptor is operating in the mouse mammary gland during pregnancy and lactation. Published 2002 Wiley-Liss, Inc.(dagger)

    DOI: 10.1002/jcp.10037

    Web of Science

    researchmap

  • Growth inhibition of mammalian cells by eosinophil cationic protein

    T Maeda, M Kitazoe, H Tada, R de Llorens, DS Salomon, M Ueda, H Yamada, M Seno

    EUROPEAN JOURNAL OF BIOCHEMISTRY   269 ( 1 )   307 - 316   2002.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BLACKWELL PUBLISHING LTD  

    Eosinophil cationic protein (ECP), one of the major components of basic granules of eosinophils, is cytotoxic to tracheal epithelium. However, the extent of this effect on other cell types has not been evaluated in vitro. In this study, we evaluated the effect of ECP on 13 mammalian cell lines. ECP inhibited the growth of several cell lines including those derived from carcinoma and leukemia in a dose-dependent manner. The IC50 values on A431 cells, MDA-MB-453 cells, HL-60 cells and K562 cells were estimated to be approximate to1-5 muM. ECP significantly suppressed the size of colonies of A431 cells, and decreased K562 cells in G(1)/G(0) phase. However, there was little evidence that ECP killed cells in either cell line. These effects of ECP were not enhanced by extending its N-terminus. Rhodamine B isothiocyanate-labeled ECP started to bind to A431 cells after 0.5 h and accumulated for up to 24 h, indicating that specific affinity for the cell surface may be important. ne affinity of ECP for heparin was assessed and found to be reduced when tryptophan residues, one of which is located at a position in the catalytic subsite of ribonuclease in ECP, were modified. The growth-inhibitory effect was also attenuated by this modification. These results suggest that growth inhibition by ECP is dependent on cell type and is cytostatic.

    DOI: 10.1046/j.0014-2956.2001.02653.x

    Web of Science

    researchmap

  • Human betacellulin structure modeled from other members of EGF family Reviewed

    Inés López-Torrejón, Enrique Querol, Francesc Avilés, Masaharu Seno, Rafael de Llorens, Baldomero Oliva

    Journal of Molecular Modeling   8 ( 4 )   131 - 144   2002

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    We have modeled betacellulin (BTC) to gain insight into the structural elements that can explain its properties. The epidermal growth factor (EGF) signal transduction pathway, a significant mediator of several cell functions, is based on four closely related tyrosine kinase receptors. The ErbB receptors are transmembrane glycoproteins and signal transduction is initiated by ligand binding that induces receptor homo- or heterodimerization to form a complex containing two molecules of ligand and two molecules of receptor. The EGF family of ligands can be divided into three groups based on their ability to bind and activate distinct ErbB receptor homo- and heterodimers. Each member of the group formed by BTC, heparin binding EGF (HB-EGF) and epiregulin (EP) can interact with both the EGF receptor (EGFR) and heregulin receptors (ErbB-3 and ErbB-4), and are hence called "bispecific" ligands. BTC and EP also present the distinctive feature that they activate all possible heterodimeric ErbB receptors. BTC has Been modeled with the program MODELLER, using human EGF, human transforming growth factor alpha (hTGFα), human HB-EGF and human heregulin one alpha (hHRG-1α) as templates. The structure of the model as well as that of the templates were optimized and a simulation of 100 ps was run for all. The main structural properties of the model and the templates were compared and in conclusion the hBTC conformation was closely similar to that of hTGFα.

    DOI: 10.1007/s00894-002-0072-2

    Scopus

    PubMed

    researchmap

  • 細胞増殖因子ベータセルリン -膵臓再生と糖尿病研究へのニューマテリアル-

    妹尾昌治

    和光純薬時報   70(3): 6-8.   2002

     More details

  • Promotion of beta-cell regeneration by betacellulin in ninety percent-pancreatectomized rats

    L Li, M Seno, H Yamada, Kojima, I

    ENDOCRINOLOGY   142 ( 12 )   5379 - 5385   2001.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ENDOCRINE SOC  

    Betacellulin is thought to promote growth and differentiation of pancreatic beta -cells. We investigated the effect of betacellulin on regeneration of pancreatic beta -cells in 90%-pancreatectomized rats. Ninety percent pancreatectomy was performed in male Wistar rats and betacellulin (0.5 mug/g body weight) or saline was administered daily for 10 d starting immediately after pancreatectomy. In pancreatectomized rats, the morning-fed plasma glucose was significantly lower and the plasma insulin concentration was significantly higher in betacellulin-treated rats than those in control rats for up to 4 wk. Thirty days after pancreatectomy, a glucose tolerance test was performed. Betacellulin reduced the plasma glucose response to ip glucose loading. In control rats, the plasma insulin concentration was significantly lower and did not respond to glucose. In contrast, the plasma insulin concentration increased slightly but significantly in betacellulin-treated rats. Thirty days after pancreatectomy, the beta -cell mass was greater and the insulin content was significantly higher in betacellulin-treated rats than those in control rats. The numbers of islet cell-like cluster and bromodeoxy uridine/insulin double-positive cells in both islet cell-like cluster and islets were significantly higher in betacellulin-treated rats. These results indicate that administration of betacellulin improves glucose metabolism by promoting beta -cell regeneration in 90%-pancreatectomized rats.

    DOI: 10.1210/en.142.12.5379

    Web of Science

    researchmap

  • Preparation of potent cytotoxic ribonucleases by cationization: Enhanced cellular uptake and decreased interaction with ribonuclease inhibitor by chemical modification of carboxyl groups

    J Futami, T Maeda, M Kitazoe, E Nukui, H Tada, M Seno, M Kosaka, H Yamada

    BIOCHEMISTRY   40 ( 25 )   7518 - 7524   2001.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    Carboxyl groups of bovine RNase A were amidated with ethylenediamine (to convert negative charges of carboxylate anions to positive ones), 2-aminoethanol (to eliminate negative charges), and taurine (to keep negative charges), respectively, by a carbodiimide reaction. Human RNase 1 was also modified with ethylenediamine. Surprisingly, the modified RNases were all cytotoxic toward 3T3-SV-40 cells despite their decreased ribonucleolytic activity. However, their enzymatic activity was not completely eliminated by the presence of excess cytosolic RNase inhibitor (RI). As for native RNase A and RNase 1 which were not cytotoxic, they were completely inactivated by RI. More interestingly, within the cytotoxic RNase derivatives. cytotoxicity correlated well with the net positive charge. RNase 1 and RNase A modified with ethylenediamine were more cytotoxic than naturally occurring cytotoxic bovine seminal RNase. An experiment using the fluorescence-labeled RNase derivatives indicated that the more cationic RNases were more efficiently adsorbed to the cells. Thus, it is suggested that the modification of carboxyl groups could change complementarity of RNase to RI and as a result endow RNase cytotoxicity and that cationization enhances the efficiency of cellular uptake of RNase so as to strengthen its cytotoxicity. The finding that an extracellular human enzyme such as RNase 1 could be effectively internalized into the cell by cationization suggests that cationization is a simple strategy for efficient delivery of a protein into cells and may open the way of the development of new therapeutics.

    DOI: 10.1021/bi010248g

    Web of Science

    researchmap

  • Physicochemical and immunological characterization of hepatitis B virus envelope particles exclusively consisting of the entire L (pre-S1+pre-S2+S) protein

    T Yamada, H Iwabuki, T Kanno, H Tanaka, T Kawai, H Fukuda, A Kondo, M Seno, K Tanizawa, S Kuroda

    VACCINE   19 ( 23-24 )   3154 - 3163   2001.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCI LTD  

    The hepatitis B virus (HBV) envelope (env) protein is composed of three regions: the 108- or 119-residue pre-Si region involved in the direct interaction with hepatocytes. the 55-residue pre-SZ region associated with the polymerized albumin-mediated interaction, and the major 226-residue S protein region. Thus, to improve the immunogenic potency of conventional HE vaccines, development of a new vaccine containing the entire pre-Si region in addition to pre-S2 and S is desired. We previously reported the efficient production of the HBV env L (pre-S1 + pre-S2 + S) protein in the recombinant yeast cells [J Biol Chem 267 (1992) 1953]. In this study, the HBV env L protein produced as nano-particles in yeast has been purified and characterized. By equilibrium sedimentation. an average molecular weight of L particle was estimated to be approximately 6.3 x 10(6), indicating that about 110 molecules of L proteins are assembled into an L particle. By atomic force microscopy in a moist atmosphere. the L particles were observed as large spherical particles with a diameter of 50-500 nm. The L particles were stable on short-time heating at a high temperature and long-time storage at a low temperature but rather unstable on repeated freezing and thawing and treatment with dithiothreitol. When immunized in mice. L particles elicited efficiently and simultaneously the anti-g, anti-pre-S2, and anti-pre-Si antibodies. The ED50 values in mice for the anti-S and anti-pre-S2 antibodies were similar to those elicited by the M (pre-S2 + S) particles. Furthermore. the anti-pre-Si rabbit antibodies were found to recognize various segments of the pre-Si region, including the pre-Si (21-47) segment. These results show the high ability of L particles to induce all antibodies against HBV env proteins, hence promising the future application of L particles for the next generation HE vaccine. (C) 2001 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0264-410X(01)00017-2

    Web of Science

    researchmap

  • Three-dimensional structure of human RNase 1 Delta N7 at 1.9 angstrom resolution

    J Pous, G Mallorqui-Fernandez, R Peracaula, SS Terzyan, J Futami, H Tada, H Yamada, M Seno, R de Llorens, FX Gomis-Ruth, M Coll

    ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY   57   498 - 505   2001.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:MUNKSGAARD INT PUBL LTD  

    Human pancreatic ribonuclease 1 (RNase 1) is considered to be the human counterpart of bovine pancreatic RNase A. Truncation of seven amino-acid residues from the aminoterminal sequence resulted in RNase 1 Delta N7, which has a reduced ribonucleolytic activity and a lower affinity for the human placental RNase inhibitor (PRI). This RNase 1 variant has been cloned, heterologously overexpressed, purified and crystallized. Its crystal structure has been determined and refined using data to 1.9 Angstrom resolution. The molecule displays the alpha + beta folding topology typical of members of the RNase A superfamily. The main distinct features found in RNase 1 Delta N7 are basically located in three loops affecting the fitting of the enzyme to the active site of subtilisin and the shape of the B2 subsite. These changes, taken with the lack of the catalytically active residue Lys7, may explain the reduced affinity of RNase 1 Delta N7 for PRI and the low ribonucleolytic activity of the protein when compared with the native enzyme.

    DOI: 10.1107/S0907444901001147

    Web of Science

    researchmap

  • Expression of phospholipid hydroperoxide glutathione peroxidase (PHGPx) mRNA in rat testes

    H Mori, T Nomura, M Seno, Y Miki, T Kimura, T Kogami, J Sasaki

    ACTA HISTOCHEMICA ET CYTOCHEMICA   34 ( 1 )   25 - 30   2001

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

    We investigated the expression of reactive oxygen species (ROS)-related enzyme mRNA to clarify the role of ROS in reproduction. In the present study, we investigated the expression of phospholipid hydroperoxide glutathione peroxidase (PHGPx) mRNA in rat testes by in situ hybridization. To prepare digoxigenin-labeled probes, we obtained 5 ' -phosphorylated oligonucleotides containing sense and antisense sequences (93-mer) with EcoR I or Hind III restriction sites as protruding cohesive ends (total 99-mer). Then, both oligonucleotides were annealed and cloned into a pGEM4Z vector. The resultant plasmid DNA was linearized with EcoR I or Hind III and used as a template for the synthesis of labeled sense or antisense riboprobes.
    PHGPx mRNA was expressed stage-specifically during spermatogenesis in adult rats. It was not expressed during spermatogonia, but its expression first appeared in stage VII pachytene spermatocytes, became evident in stage VIII pachytene spermatocytes and increased gradually until becoming diplotene spermatocytes. It decreased slightly during the metaphase of meiosis, and then gradually increased as the spermatids differentiated. The expression was marked in spermatids between steps 7 and 12, and the maximum expression was observed in step 9-10 spermatids. Expression was diminished completely in step 18 spermatids. These results suggest that PHGPx plays an essential role in spermatogenesis.

    Web of Science

    researchmap

  • ベータセルリン:細胞増殖分化因子を応用した再生医療への試み

    妹尾昌治

    ハイテクインフォメーション   134号14-21   2001

     More details

  • The EGF-CFC family: novel epidermal growth factor-related proteins in development and cancer

    DS Salomon, C Bianco, AD Ebert, NI Khan, M De Santis, N Normanno, C Wechselberger, M Seno, K Williams, M Sanicola, S Foley, W Gullick, G Persico

    ENDOCRINE-RELATED CANCER   7 ( 4 )   199 - 226   2000.12

     More details

    Language:English   Publisher:SOC ENDOCRINOLOGY  

    The EGF-CFC gene family encodes a group of structurally related proteins that serve as important competence factors during early embryogenesis in Xenopus, zebrafish, mice and humans. This multigene family consists of Xenopus FRL-1, zebrafish one-eyed-pinhead (oep), mouse cripto (Cr-l) and cryptic, and human cripto (CR-1) and criptin. FRL-1, oep and mouse cripto are essential for the formation of mesoderm and endoderm and for correct establishment of the anterior/posterior axis. In addition, oep and cryptic are important for the establishment of left-right (UR) asymmetry, In zebrafish, there is strong genetic evidence that oep functions as an obligatory cc-factor for the correct signaling of a transforming growth factor-beta (TGF beta)-related gene, nodal, during gastrulation and during UR asymmetry development. Expression of Cr-l and cryptic is extinguished in the embryo after day 8 of gestation except for the developing heart where Cr-l expression is necessary for myocardial development. In the mouse, cryptic is not expressed in adult tissues whereas Cr-l is expressed at a low level in several different tissues including the mammary gland, In the mammary gland, expression of Cr-l in the ductal epithelial cells increases during pregnancy and lactation and immunoreactive and biologically active Cr-l protein can be detected in human milk. Overexpression of Cr-l in mouse mammary epithelial cells can facilitate their in vitro transformation and in vivo these Cr-l-transduced cells produce ductal hyperplasias in the mammary gland. Recombinant mouse or human cripto can enhance cell motility and branching morphogenesis in mammary epithelial cells and in some human tumor cells. These effects are accompanied by an epithelial-mesenchymal transition which is associated with a decrease in beta -catenin function and an increase in vimentin expression. Expression of cripto is increased several-fold in human colon, gastric, pancreatic and lung carcinomas and in a variety of different types of mouse and human breast carcinomas. More importantly, this increase can first be detected in premalignant lesions in some of these tissues. Although a specific receptor for the EGF-CFC proteins has not yet been identified, oep depends upon an activin-type RIIB and RIB receptor system that functions through Smad-e. Mouse and human cripto have been shown to activate a ras/raf/MAP kinase signaling pathway in mammary epithelial cells. Activation of phosphatidylinositol 3-kinase and Akt are also important for the ability of CR-1 to stimulate cell migration and to block lactogenic hormone-induced expression of p-casein and whey acidic protein. In mammary epithelial cells, part of these responses may depend on the ability of CR-1 to transactivate erb B-4 and/or fibroblast growth factor receptor 1 through an src-like tyrosine kinase.

    DOI: 10.1677/erc.0.0070199

    Web of Science

    researchmap

  • 上皮増殖因子受容体(EGFR)過剰発現癌に対する生理活性物質を用いた低免疫原性ターゲッテイング療法の開発

    神野 浩光, 上田 政和, 菊池 潔, 池田 正, 北島 政樹, 二見 淳一郎, 妹尾 昌治

    日本外科学会雑誌   101 ( 臨増 )   411 - 411   2000.3

     More details

    Language:Japanese   Publisher:(一社)日本外科学会  

    researchmap

  • Processing and juxtacrine activity of membrane-anchored betacellulin

    H Tada, R Sasada, Y Kawaguchi, Kojima, I, WJ Gullick, DS Salomon, K Igarashi, M Seno, H Yamada

    JOURNAL OF CELLULAR BIOCHEMISTRY   72 ( 3 )   423 - 434   1999.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-LISS  

    Betacellulin (BTC) was originally isolated as a secreted growth factor from a mouse pancreatic beta-tumor cell line, whereas the cDNA sequence predicts that ETC is synthesized as a larger transmembrane protein. In the present study, we have characterized the membrane-anchored forms of BTC, using Chinese hamster ovary (CHO) cells, mouse fibroblast A9 cells, and a human breast cancer cell line MCF-7, all of which were stably transfected with human ETC cDNA. A9 and MCF-7 transfectants produced membrane-anchored ETC isoforms of 21, 25, 29, and 40 kDa on the cell surface, as well as a secreted BTC isoform. CHO transfectants secreted little ETC but accumulated the membrane-anchored isoforms. The cleavage of the membrane-anchored forms to release a secreted from of ETC was not enhanced by biological mediators such as a phorbol ester, which stimulates the cleavage of other membrane-anchored growth factors. The membrane-anchored forms of ETC expressed on the transfected cells induced the insulin production and/or promoted the growth in subclones of AR42J rat pancreatic cells. These results suggest that the membrane-anchored ETC can function as a juxtacrine factor in regulating the growth and differentiation of pancreatic endocrine cells. J. Cell. Biochem. 72.423-134, 1999. (C) 1999 Wiley-Liss, Inc.

    DOI: 10.1002/(SICI)1097-4644(19990301)72:3<423::AID-JCB11>3.0.CO;2-P

    Web of Science

    researchmap

  • The three-dimensional structure of human RNase 4, unliganded and complexed with d(Up), reveals the basis for its uridine selectivity

    SS Terzyan, R Peracaula, R de Llorens, Y Tsushima, H Yamada, M Seno, FX Gomis-Ruth, M Coll

    JOURNAL OF MOLECULAR BIOLOGY   285 ( 1 )   205 - 214   1999.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS LTD  

    The RNase 4 family is unique among RNase enzymes, displaying the highest level of sequence similarity and encompassing the shortest polypeptide chain. It is the only one showing high specificity. The human representative is an intracellular and plasma enzyme, first isolated from colon adenocarcinoma cell line HT-29. The crystal structures of human recombinant RNase 4, unliganded and in complex with d(Up), have been determined, revealing in the unique active site an explanation for the uridine specificity. Arg101, at a position not involved in catalysis in the other RNase enzymes, penetrates the enzyme moiety shaping the recognition pocket, a flip that is mediated by the interaction with the (shorter chain) C-terminal carboxylate group, providing an anchoring point for the O4 atom of the substrate uridine. The bulky Phe42 side-chain forces Asp80 to be in the chi(1) = -72.49 degrees rotamer, accepting a hydrogen bond from Thr44, further converting the latter into a hydrogen bond acceptor. This favours an interaction with the -NH-donor group of uridine at position 3 over that with the =N-acceptor of cytidine. The two chemical groups that distinguish uracyl from cytosine are used by the enzyme to discriminate between these two bases. (C) 1999 Academic Press.

    DOI: 10.1006/jmbi.1998.2288

    Web of Science

    researchmap

  • 新しい臓器特異的癌関連遺伝子のクローニングと癌遺伝子産物を分子標的とした治療法の開発

    上田 政和, 菊池 潔, 板野 理, プサラス・キリヤコス, 北島 政樹, 妹尾 昌治

    日本癌治療学会誌   33 ( 3 )   355 - 355   1998.8

     More details

    Language:Japanese   Publisher:(一社)日本癌治療学会  

    researchmap

  • Identification of the genes associated with differentiation of AR42J cells into insulin-secreting cells

    H Mashima, J Takeda, M Seno, H Yamada, Kojima, I

    DIABETES   47   A176 - A176   1998.5

     More details

    Language:English   Publisher:AMER DIABETES ASSOC  

    Web of Science

    researchmap

  • Molecular targeting for activated T cell by recombinant human RNase and IL-2.

    M Ueda, K Psarras, M Tanabe, T Yamamura, M Kitajima, M Seno, S Komatsu

    GASTROENTEROLOGY   114 ( 4 )   A1186 - A1186   1998.4

     More details

    Language:English   Publisher:W B SAUNDERS CO  

    Web of Science

    researchmap

  • Characterization of the betacellulin receptor in pancreatic AR42J-B20 cells

    N Ishiyama, M Kanzaki, M Furukawa, J Miyagawa, T Hanafusa, M Seno, H Yamada, Kobayashi, I, Kojima, I

    DIABETOLOGIA   40   91 - 91   1997.6

     More details

    Language:English   Publisher:SPRINGER VERLAG  

    Web of Science

    researchmap

  • Genes expressed during the differentiation of AR42J cells into insulin-secreting

    H Mashima, J Takeda, M Seno, H Yamada, Kojima, I

    DIABETES   46   1378 - 1378   1997.5

     More details

    Language:English   Publisher:AMER DIABETES ASSOC  

    Web of Science

    researchmap

  • Molecular Targeting for Epidermal Growth Factor Receptor Expressed on Breast Cancer Cells by Human Fusion Protein Reviewed

    Ueda M, Psarras K, Jinno H, Ikeda T, Enomoto K, Kitajima M, Futami J, Yamada H, Seno M

    Breast Cancer   4 ( 4 )   253 - 255   1997

     More details

▼display all

Books

  • 次世代のがん治療薬・診断のための研究開発~免疫療法・遺伝子治療・がん幹細胞~、第13章 がん幹細胞を標的とした治療薬研究 iPS細胞を用いたがん幹細胞モデル作成のポイントとその評価

    2016 

     More details

  • 遺伝子・組織・生物活性化合物のクラスタリング

    海文堂出版  2015 

     More details

  • Insight into Cancer Stem Cell Niche; Lessons from Cancer Stem Cell Models Generated In Vitro. In: Biology in Stem Cell Niche (Ed: Kursad Turksen)

    Humana Press  2015 

     More details

  • がん幹細胞から考えるがんの正体 —クローン説に基づく治療薬/治療法からのパラダイムシフト

    化学同人社  2014 

     More details

  • マウスiPS細胞から作るがん幹細胞モデル

    羊土社  2013 

     More details

  • Exploring the mechanism for biological evolution: DNA methyltransferase is the pushing power of DNA and protein evolution.

    Lambert Academic Publishing  2011 

     More details

  • Self Organizing Maps - Applications and Novel Algorithm Design

    InTech  2011 

     More details

  • 中空バイオナノ粒子を用いるピンポイントDDSおよび遺伝子導入法

    シーエムシー出版(東京)  2009 

     More details

  • Pinpoint DDS and gene transduction using hollow bio-nanoparticle

    2009 

     More details

  • 自己組織化マップとそのツール

    シュプリンガー・ジャパン  2008 

     More details

  • Chapter 11 Protein Sequence Analysis; in “Bioinformatics – a practical approach- ed.Shui Qing Ye, (Mathematical and Computational Biology Series)”

    Chapman & Hall/CRC  2007 

     More details

  • 自己組織化マップとその応用

    シュプリンガー・ジャパン  2007 

     More details

  • Bioinformatics - a practical approach -

    Chapman & Hall/CRC  2007 

     More details

  • ナノパーティクル・テクノロジー

    日刊工業新聞社(東京)  2003 

     More details

  • 「中空バイオナノ粒子を 用いるピンポイントDDSおよび遺伝子導入法」ナノバイオテクノロジー最前線

    シ ーエムシー出版(東京)  2003 

     More details

  • 生物工学系テキストシリーズ「バイオ機器分析入門」

    講談社サイエンティフィク  2000 

     More details

  • FGF-7(KGF)

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • 線維芽細胞成長因子ファミリーFGF

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • FGF-1(酸性線維芽細胞成長因子,aFGF)

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • FGF-2(塩基性線維芽細胞成長因子,bFGF)

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • FGF-3(int-2遺伝子産物)

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • FGF-4(hst-1/kFGF遺伝子産物)

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • FGF-5

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • FGF-6(hst-2遺伝子産物)

    タンパク質化学第8巻「成長因子I」広川書店 

     More details

  • 生物工学系テキストシリーズ「バイオ機能分析入門」、「第2章 クロマトグラフィー」

    講談社サイエンティフィク 

     More details

▼display all

MISC

  • HSP-enriched properties of extracellular vesicles involve survival of metastatic oral cancer cells. Reviewed International journal

    Kisho Ono, Takanori Eguchi, Chiharu Sogawa, Stuart K Calderwood, Junya Futagawa, Tomonari Kasai, Masaharu Seno, Kuniaki Okamoto, Akira Sasaki, Ken-Ichi Kozaki

    Journal of cellular biochemistry   119 ( 9 )   7350 - 7362   2018.9

     More details

    Language:English  

    Cancer cells often secrete extracellular vesicles (EVs) that carry heat shock proteins (HSPs) with roles in tumor progression. Oral squamous cell carcinoma (OSCC) belongs to head and neck cancers (HNC) whose lymph-node-metastases often lead to poor prognosis. We have examined the EV proteome of OSCC cells and found abundant secretion of HSP90-enriched EVs in lymph-node-metastatic OSCC cells. Double knockdown of HSP90α and HSP90β, using small interfering RNA significantly reduced the survival of the metastatic OSCC cells, although single knockdown of each HSP90 was ineffective. Elevated expression of these HSP90 family members was found to correlate with poor prognosis of HNC cases. Thus, elevated HSP90 levels in secreted vesicles are potential prognostic biomarkers and therapeutic targets in metastatic OSCC.

    DOI: 10.1002/jcb.27039

    PubMed

    researchmap

  • Up-Regulation of PI 3-Kinases and the Activation of PI3K-Akt Signaling Pathway in Cancer Stem-Like Cells Through DNA Hypomethylation Mediated by the Cancer Microenvironment. Reviewed International journal

    Aung Ko Ko Oo, Anna Sanchez Calle, Neha Nair, Hafizah Mahmud, Arun Vaidyanath, Junya Yamauchi, Aprilliana Cahya Khayrani, Juan Du, Md Jahangir Alam, Akimasa Seno, Akifumi Mizutani, Hiroshi Murakami, Yoshiaki Iwasaki, Ling Chen, Tomonari Kasai, Masaharu Seno

    Translational oncology   11 ( 3 )   653 - 663   2018.6

     More details

    Language:English  

    Previously, we have succeeded in converting induced pluripotent stem cells (iPSCs) into cancer stem cells (CSCs) by treating the iPSCs with conditioned medium of Lewis lung carcinoma (LLC) cells. The converted CSCs, named miPS-LLCcm cells, exhibited the self-renewal, differentiation potential, and potential to form malignant tumors with metastasis. In this study, we further characterized miPS-LLCcm cells both in vivo and in vitro. The tumors formed by subcutaneous injection showed the structures with pathophysiological features consisting of undifferentiated and malignant phenotypes generally found in adenocarcinoma. Metastasis in the lung was also observed as nodule structures. Excising from the tumors, primary cultured cells from the tumor and the nodule showed self-renewal, differentiation potential as well as tumor forming ability, which are the essential characters of CSCs. We then characterized the epigenetic regulation occurring in the CSCs. By comparing the DNA methylation level of CG rich regions, the differentially methylated regions (DMRs) were evaluated in all stages of CSCs when compared with the parental iPSCs. In DMRs, hypomethylation was found superior to hypermethylation in the miPS-LLCcm cells and its derivatives. The hypo- and hypermethylated genes were used to nominate KEGG pathways related with CSC. As a result, several categories were defined in the KEGG pathways from which most related with cancers, significant and high expression of components was PI3K-AKT signaling pathway. Simultaneously, the AKT activation was also confirmed in the CSCs. The PI3K-Akt signaling pathway should be an important pathway for the CSCs established by the treatment with conditioned medium of LLC cells.

    DOI: 10.1016/j.tranon.2018.03.001

    PubMed

    researchmap

  • Cytokine Expression and Macrophage Localization in Xenograft and Allograft Tumor Models Stimulated with Lipopolysaccharide. Reviewed International journal

    Junko Masuda, Tsukasa Shigehiro, Takuma Matsumoto, Ayano Satoh, Akifumi Mizutani, Chiho Umemura, Shoki Saito, Mayumi Kijihira, Eiji Takayama, Akimasa Seno, Hiroshi Murakami, Masaharu Seno

    International journal of molecular sciences   19 ( 4 )   2018.4

     More details

    Language:English  

    T cell-deficient mice such as nude mice are often used to generate tumor xenograft for the development of anticancer agents. However, the functionality of the other immune cells including macrophages, dendritic cells (DCs), and myeloid-derived suppressor cells (MDSCs) in the xenograft are largely unknown. Macrophages and dendritic cells (DCs) acquire functionally distinct properties in response to various environmental stimuli; the interaction of these cells with MDSCs in tumor microenvironments regulates cancer progression. Nude mice are less likely to reject human cancer cells because of major histocompatibility complex (MHC) mismatches. The tumor microenvironment in a xenograft, comprising human and mouse cells, exhibits more complex bidirectional signaling and function than that of allograft. Here, we evaluated the differences of myeloid cells between them. Plasma interferon-&gamma; and interleukin-18 concentrations in the xenograft tumor model after lipopolysaccharide (LPS) administration were significantly higher than those in the allograft tumor model. MHC class I, II, and CD80 expression levels were increased in CD11b⁺ and MDSC populations after LPS administration in the spleen of a xenograft tumor model but not in that of an allograft tumor model. Additionally, the number of CD80- and mannose receptor C type 1 (MRC1)-expressing cells was decreased upon LPS administration in the tumor of the xenograft tumor. These results suggest that functions of macrophages and DCs are sustained in the xenograft, whereas their functions in response to LPS were suppressed in the allograft. The findings will encourage the consideration of the effects of myeloid cells in the xenograft for drug development.

    DOI: 10.3390/ijms19041261

    PubMed

    researchmap

  • Targeting Glioblastoma Cells Expressing CD44 with Liposomes Encapsulating Doxorubicin and Displaying Chlorotoxin-IgG Fc Fusion Protein. Reviewed International journal

    Hafizah Mahmud, Tomonari Kasai, Apriliana Cahya Khayrani, Mami Asakura, Aung Ko Ko Oo, Juan Du, Arun Vaidyanath, Samah El-Ghlban, Akifumi Mizutani, Akimasa Seno, Hiroshi Murakami, Junko Masuda, Masaharu Seno

    International journal of molecular sciences   19 ( 3 )   2018.2

     More details

    Language:English  

    We recently have established a successful xenograft model of human glioblastoma cells by enriching hyaluronic acid-dependent spheroid-forming populations termed U251MG-P1 cells from U251MG cells. Since U251MG-P1 cells have been confirmed to express CD44 along with principal stemness marker genes, OCT3/4, SOX2, KLF4 and Nanog, this CD44 expressing population appeared to majorly consist of undifferentiated cells. Evaluating the sensitivity to anti-cancer agents, we found U251MG-P1 cells were sensitive to doxorubicin with IC50 at 200 nM. Although doxorubicin has serious side-effects, establishment of an efficient therapy targeting undifferentiated glioblastoma cell population is necessary. We previously designed a chlorotoxin peptide fused to human IgG Fc region without hinge sequence (M-CTX-Fc), which exhibited a stronger growth inhibitory effect on the glioblastoma cell line A172 than an original chlorotoxin peptide. Combining these results together, we designed M-CTX-Fc conjugated liposomes encapsulating doxorubicin and used U251MG-P1 cells as the target model in this study. The liposome modified with M-CTX-Fc was designed with a diameter of approximately 100-150 nm and showed high encapsulation efficiency, adequate loading capacity of anticancer drug, enhanced antitumor effects demonstrating increasing uptake into the cells in vitro; M-CTX-Fc-L-Dox shows great promise in its ability to suppress tumor growth in vivo and it could serve as a template for targeted delivery of other therapeutics.

    DOI: 10.3390/ijms19030659

    PubMed

    researchmap

  • Iron depletion is a novel therapeutic strategy to target cancer stem cells. Reviewed International journal

    Takayuki Ninomiya, Toshiaki Ohara, Kazuhiro Noma, Yuki Katsura, Ryoichi Katsube, Hajime Kashima, Takuya Kato, Yasuko Tomono, Hiroshi Tazawa, Shunsuke Kagawa, Yasuhiro Shirakawa, Fumiaki Kimura, Ling Chen, Tomonari Kasai, Masaharu Seno, Akihiro Matsukawa, Toshiyoshi Fujiwara

    Oncotarget   8 ( 58 )   98405 - 98416   2017.11

     More details

    Language:English  

    Adequate iron levels are essential for human health. However, iron overload can act as catalyst for the formation of free radicals, which may cause cancer. Cancer stem cells (CSCs), which maintain the hallmark stem cell characteristics of self-renewal and differentiation capacity, have been proposed as a driving force of tumorigenesis and metastases. In the present study, we investigated the role of iron in the proliferation and stemness of CSCs, using the miPS-LLCcm cell model. Although the anti-cancer agents fluorouracil and cisplatin suppressed the proliferation of miPS-LLCcm cells, these drugs did not alter the expression of stemness markers, including Nanog, SOX2, c-Myc, Oct3/4 and Klf4. In contrast, iron depletion by the iron chelators deferasirox and deferoxamine suppressed the proliferation of miPS-LLCcm cells and the expression of stemness markers. In an allograft model, deferasirox inhibited the growth of miPS-LLCcm implants, which was associated with decreased expression of Nanog and Sox2. Altogether, iron appears to be crucial for the proliferation and maintenance of stemness of CSCs, and iron depletion may be a novel therapeutic strategy to target CSCs.

    DOI: 10.18632/oncotarget.21846

    PubMed

    researchmap

  • Practical Liposomal Formulation for Taxanes with Polyethoxylated Castor Oil and Ethanol with Complete Encapsulation Efficiency and High Loading Efficiency. Reviewed International journal

    Tsukasa Shigehiro, Junko Masuda, Shoki Saito, Apriliana C Khayrani, Kazumasa Jinno, Akimasa Seno, Arun Vaidyanath, Akifumi Mizutani, Tomonari Kasai, Hiroshi Murakami, Ayano Satoh, Tetsuya Ito, Hiroki Hamada, Yuhki Seno, Tadakatsu Mandai, Masaharu Seno

    Nanomaterials (Basel, Switzerland)   7 ( 10 )   1391 - 1398   2017.9

     More details

    Language:English  

    Taxanes including paclitaxel and docetaxel are effective anticancer agents preferably sufficient for liposomal drug delivery. However, the encapsulation of these drugs with effective amounts into conventional liposomes is difficult due to their high hydrophobicity. Therefore, an effective encapsulation strategy for liposomal taxanes has been eagerly anticipated. In this study, the mixture of polyethoxylated castor oil (Cremophor EL) and ethanol containing phosphate buffered saline termed as CEP was employed as a solvent of the inner hydrophilic core of liposomes where taxanes should be incorporated. Docetaxel-, paclitaxel-, or 7-oxacetylglycosylated paclitaxel-encapsulating liposomes were successfully prepared with almost 100% of encapsulation efficiency and 29.9, 15.4, or 29.1 mol% of loading efficiency, respectively. We then applied the docetaxel-encapsulating liposomes for targeted drug delivery. Docetaxel-encapsulating liposomes were successfully developed HER2-targeted drug delivery by coupling HER2-specific binding peptide on liposome surface. The HER2-targeting liposomes exhibited HER2-specific internalization and enhanced anticancer activity in vitro. Therefore, we propose the sophisticated preparation of liposomal taxanes using CEP as a promising formulation for effective cancer therapies.

    DOI: 10.3390/nano7100290

    PubMed

    researchmap

  • Synthesis and assembly of Hepatitis B virus envelope protein-derived particles in Escherichia coli. Reviewed International journal

    Hao Li, Keisuke Onbe, Qiushi Liu, Masumi Iijima, Kenji Tatematsu, Masaharu Seno, Hiroko Tada, Shun' Ichi Kuroda

    Biochemical and biophysical research communications   490 ( 2 )   155 - 160   2017.8

     More details

    Language:English  

    Hepatitis B virus (HBV) envelope particles have been synthesized in eukaryotic cells (e.g., mammalian cells, insect cells, and yeast cells) as an HB vaccine immunogen and drug delivery system (DDS) nanocarrier. Many researchers had made attempts to synthesize the particles in Escherichia coli for minimize the cost and time for producing HBV envelope particles, but the protein was too deleterious to be synthesized in E. coli. In this study, we generated deletion mutants of HBV envelope L protein (389 amino acid residues (aa)) containing three transmembrane domains (TM1, TM2, TM3). The ΔNC mutant spanning from TM2 to N-terminal half of TM3 (from 237 aa to 335 aa) was found as a shortest form showing spontaneous particle formation. After the N-terminal end of ΔNC mutant was optimized by the N-end rule for E. coli expression, the modified ΔNC mutant (mΔNC) was efficiently expressed as particles in E. coli. The molecular mass of mΔNC particle was approx. 670 kDa, and the diameter was 28.5 ± 6.2 nm (mean ± SD, N = 61). The particle could react with anti-HBV envelope S protein antibody, indicating the particles exhibited S antigenic domain outside as well as HBV envelope particles. Taken together, the E. coli-derived mΔNC particles could be used as a substitute of eukaryotic cell-derived HBV envelope particles for versatile applications.

    DOI: 10.1016/j.bbrc.2017.06.015

    PubMed

    researchmap

  • A cancer stem cell model as the point of origin of cancer-associated fibroblasts in tumor microenvironment. Reviewed International journal

    Neha Nair, Anna Sanchez Calle, Maram Hussein Zahra, Marta Prieto-Vila, Aung Ko Ko Oo, Laura Hurley, Arun Vaidyanath, Akimasa Seno, Junko Masuda, Yoshiaki Iwasaki, Hiromi Tanaka, Tomonari Kasai, Masaharu Seno

    Scientific reports   7 ( 1 )   6838 - 6838   2017.7

     More details

    Language:English  

    Cancer-associated fibroblasts (CAFs) are one of the most prominent cell types in the stromal compartment of the tumor microenvironment. CAFs support multiple aspects of cancer progression, including tumor initiation, invasion, and metastasis. The heterogeneous nature of the stromal microenvironment is attributed to the multiple sources from which the cells in this compartment originate. The present study provides the first evidence that cancer stem cells (CSCs) are one of the key sources of CAFs in the tumor niche. We generated CSC-like cells by treating mouse induced pluripotent stem cells with conditioned medium from breast cancer cell lines. The resulting cell population expressed both CSC and pluripotency markers, and the sphere-forming CSC-like cells formed subcutaneous tumors in nude mice. Intriguingly, these CSC-like cells always formed heterogeneous populations surrounded by myofibroblast-like cells. Based on this observation, we hypothesized that CSCs could be the source of the CAFs that support tumor maintenance and survival. To address this hypothesis, we induced the differentiation of spheres and purified the myofibroblast-like cells. The resulting cells exhibited a CAF-like phenotype, suggesting that they had differentiated into the subpopulations of cells that support CSC self-renewal. These findings provide novel insights into the dynamic interplay between various microenvironmental factors and CAFs in the CSC niche.

    DOI: 10.1038/s41598-017-07144-5

    PubMed

    researchmap

  • Tumor growth limited to subcutaneous site vs tumor growth in pulmonary site exhibit differential effects on systemic immunities. Reviewed International journal

    Junko Masuda, Eiji Takayama, Warren Strober, Ayano Satoh, Yuji Morimoto, Yasuko Honjo, Tatsuo Ichinohe, Shin-Ichi Tokuno, Toshiaki Ishizuka, Takahiro Nakata, Akifumi Mizutani, Naoki Umemura, Atsushi Kitani, Ivan J Fuss, Tsukasa Shigehiro, Harumi Kawaki, Masako Mizuno-Kamiya, Nobuo Kondoh, Masaharu Seno

    Oncology reports   38 ( 1 )   449 - 455   2017.7

     More details

    Language:English  

    To evaluate systemic immunity associated with tumor growth limited to a subcutaneous site versus growth proceeding at multiple tumor sites, we established syngeneic mouse subcutaneous and pulmonary tumor models by local subcutaneous and intravenous injection of colon carcinoma CT26 cells. We found that splenic myeloid-derived suppressor cell (MDSC) levels were significantly increased in the subcutaneous tumor model but not in the pulmonary tumor model. Furthermore, both CD4+ and CD8+ T cells as well as CD4+ Foxp3+ T cells were significantly decreased in the subcutaneous tumor model and were largely unchanged in the pulmonary tumor model. In addition, the subcutaneous model, but not the pulmonary model, displayed a Th1 polarization bias. This bias was characterized by decreased IL-4, IL-9, and IL-10 production, whereas the pulmonary model displayed increased production of IL-10. These results suggest that the mode of tumor development has differential effects on systemic immunity that may, in turn, influence approaches to treatment of cancer patients.

    DOI: 10.3892/or.2017.5646

    PubMed

    researchmap

  • Anticancer effects of novel thalidomide analogs in A549 cells through inhibition of vascular endothelial growth factor and matrix metalloproteinase-2

    Bishoy El-Aarag, Tomonari Kasai, Junko Masuda, Hussein Agwa, Magdy Zahran, Masaharu Seno

    BIOMEDICINE & PHARMACOTHERAPY   85   549 - 555   2017.1

     More details

    Language:English   Publisher:ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER  

    Lung cancer is one of the major causes of cancer-related mortality worldwide, and non-small-cell lung cancer is the most common form of lung cancer. Several studies had shown that thalidomide has potential for prevention and therapy of cancer. Therefore, the current study aimed to investigate the antitumor effects of two novel thalidomide analogs in human lung cancer A549 cells. The antiproliferative, antimigratory, and apoptotic effects in A549 cells induced by thalidomide analogs were examined. In addition, their effects on the expression of mRNAs encoding vascular endothelial growth factor165 (VEGF165) and matrix metalloproteinase-2 (MMP-2) were evaluated. Their influence on the tumor volume in nude mice was also determined. Results revealed that thalidomide analogs exhibited antiproliferative, antimigratory, and apoptotic activities with more pronounced effect than thalidomide drug. Furthermore, analogs 1 and 2 suppressed the expression levels of VEGF165 by 42% and 53.2% and those of MMP-2 by 45% and 52%, respectively. Thalidomide analogs 1 and 2 also reduced the tumor volume by 30.11% and 53.52%, respectively. Therefore, this study provides evidence that thalidomide analogs may serve as a new therapeutic option for treating lung cancer. (C) 2016 Elsevier Masson SAS. All rights reserved.

    DOI: 10.1016/j.biopha.2016.11.063

    Web of Science

    researchmap

  • Anticancer effects of novel thalidomide analogs in A549 cells through inhibition of vascular endothelial growth factor and matrix metalloproteinase-2

    Bishoy El-Aarag, Tomonari Kasai, Junko Masuda, Hussein Agwa, Magdy Zahran, Masaharu Seno

    BIOMEDICINE & PHARMACOTHERAPY   85   549 - 555   2017.1

     More details

    Authorship:Lead author   Language:English   Publisher:ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER  

    Lung cancer is one of the major causes of cancer-related mortality worldwide, and non-small-cell lung cancer is the most common form of lung cancer. Several studies had shown that thalidomide has potential for prevention and therapy of cancer. Therefore, the current study aimed to investigate the antitumor effects of two novel thalidomide analogs in human lung cancer A549 cells. The antiproliferative, antimigratory, and apoptotic effects in A549 cells induced by thalidomide analogs were examined. In addition, their effects on the expression of mRNAs encoding vascular endothelial growth factor165 (VEGF165) and matrix metalloproteinase-2 (MMP-2) were evaluated. Their influence on the tumor volume in nude mice was also determined. Results revealed that thalidomide analogs exhibited antiproliferative, antimigratory, and apoptotic activities with more pronounced effect than thalidomide drug. Furthermore, analogs 1 and 2 suppressed the expression levels of VEGF165 by 42% and 53.2% and those of MMP-2 by 45% and 52%, respectively. Thalidomide analogs 1 and 2 also reduced the tumor volume by 30.11% and 53.52%, respectively. Therefore, this study provides evidence that thalidomide analogs may serve as a new therapeutic option for treating lung cancer. (C) 2016 Elsevier Masson SAS. All rights reserved.

    DOI: 10.1016/j.biopha.2016.11.063

    Web of Science

    researchmap

  • Hyaluronic Acid Mediated Enrichment of CD44 Expressing Glioblastoma Stem Cells in U251MG Xenograft Mouse Model.

    Vaidyanath A, Mahmud HB, Khayrani AC, Oo AK, Seno A, Asakura A, Kasai T, Seno M

    Journal of Stem Cell Research and Therapy   2017

  • Immunoliposomes: Recent Progress of Liposomal Drug Delivery System Coupled with Antibodies. in “Liposomes: Historical, Clinical and Molecular Perspectives

    Shigehiro T, Seno, M

    Liposomes Historical,Clinical and Molecular Perspectives   189 - 209   2017

     More details

  • 腫瘍を標的とした抗体提示リポソームの開発と製剤化

    妹尾昌治, 重廣司

    DDS先端技術の製剤への応用開発   249 - 262   2017

     More details

  • Transient Tcf3 Gene Repression by TALE-Transcription Factor Targeting

    Junko Masuda, Hiroshi Kawamoto, Warren Strober, Eiji Takayama, Akifumi Mizutani, Hiroshi Murakami, Tomokatsu Ikawa, Atsushi Kitani, Narumi Maeno, Tsukasa Shigehiro, Ayano Satoh, Akimasa Seno, Vaidyanath Arun, Tomonari Kasai, Ivan J. Fuss, Yoshimoto Katsura, Masaharu Seno

    APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY   180 ( 8 )   1559 - 1573   2016.12

     More details

    Language:English   Publisher:HUMANA PRESS INC  

    Transplantation of hematopoietic stem and progenitor cells (HSCs) i.e., self-renewing cells that retain multipotentiality, is now a widely performed therapy for many hematopoietic diseases. However, these cells are present in low number and are subject to replicative senescence after extraction; thus, the acquisition of sufficient numbers of cells for transplantation requires donors able to provide repetitive blood samples and/or methods of expanding cell numbers without disturbing cell multipotentiality. Previous studies have shown that HSCs maintain their multipotentiality and self-renewal activity if TCF3 transcription function is blocked under B cell differentiating conditions. Taking advantage of this finding to devise a new approach to HSC expansion in vitro, we constructed an episomal expression vector that specifically targets and transiently represses the TCF3 gene. This consisted of a vector encoding a transcription activator-like effector (TALE) fused to a Kruppel-associated box (KRAB) repressor. We showed that this TALE-KRAB vector repressed expression of an exogenous reporter gene in HEK293 and COS-7 cell lines and, more importantly, efficiently repressed endogenous TCF3 in a human B lymphoma cell line. These findings suggest that this vector can be used to maintain multipotentiality in HSC being subjected to a long-term expansion regimen prior to transplantation.

    DOI: 10.1007/s12010-016-2187-4

    Web of Science

    researchmap

  • Meibomian gland dysfunction model in hairless mice bred under special diet limiting lipid content

    Hideki Miyake, Tomoko Oda, Osamu Katsuta, Masaharu Seno, Masatsugu Nakamura

    INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE   57 ( 12 )   2016.9

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:ASSOC RESEARCH VISION OPHTHALMOLOGY INC  

    Web of Science

    researchmap

  • Characterization of gene expression patterns among artificially developed cancer stem cells using spherical self-organizing map

    Akimasa Seno, Tomonari Kasai, Masashi Ikeda, Arun Vaidyanath, Junko Masuda, Akifumi Mizutani, Hiroshi Murakami, Tetsuya Ishikawa, Masaharu Seno

    Cancer Informatics   15   163 - 178   2016.8

     More details

    Language:English   Publisher:Libertas Academica Ltd.  

    We performed gene expression microarray analysis coupled with spherical self-organizing map (sSOM) for artificially developed cancer stem cells (CSCs). The CSCs were developed from human induced pluripotent stem cells (hiPSCs) with the conditioned media of cancer cell lines, whereas the CSCs were induced from primary cell culture of human cancer tissues with defined factors (OCT3/4, SOX2, and KLF4). These cells commonly expressed human embryonic stem cell (hESC)/hiPSC-specific genes (POU5F1, SOX2, NANOG, LIN28, and SALL4) at a level equivalent to those of control hiPSC 201B7. The sSOM with unsupervised method demonstrated that the CSCs could be divided into three groups based on their culture conditions and original cancer tissues. Furthermore, with supervised method, sSOM nominated TMED9, RNASE1, NGFR, ST3GAL1, TNS4, BTG2, SLC16A3, CD177, CES1, GDF15, STMN2, FAM20A, NPPB, CD99, MYL7, PRSS23, AHNAK, and LOC152573 genes commonly upregulating among the CSCs compared to hiPSC, suggesting the gene signature of the CSCs.

    DOI: 10.4137/CIN.S39839

    Scopus

    researchmap

  • Cancer stem cells as the novel origin of cancer-associated fibroblast-like cells

    Neha Nair, Arun Vaidyanath, Kenta Hoshikawa, Anna Sanchez Calle, Tomonari Kasai, Masaharu Seno

    CANCER RESEARCH   76   2016.7

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2016-LB-278

    Web of Science

    researchmap

  • The significance of c-Kit protooncogene in iCSC-derived PDAC model

    Anna Sanchez Calle, Kenta Hoshikawa, Neha Nair, Marta Prieto-Vila, Arun Vaidyanath, Tomonari Kasai, Masaharu Seno

    CANCER RESEARCH   76   2016.7

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2016-1725

    Web of Science

    researchmap

  • Meibomian Gland Dysfunction Model in Hairless Mice Fed a Special Diet With Limited Lipid Content

    Hideki Miyake, Tomoko Oda, Osamu Katsuta, Masaharu Seno, Masatsugu Nakamura

    INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE   57 ( 7 )   3268 - 3275   2016.6

     More details

    Language:English   Publisher:ASSOC RESEARCH VISION OPHTHALMOLOGY INC  

    PURPOSE. A novel meibomian gland dysfunction (MGD) model was developed to facilitate understanding of the pathophysiology of MGD and to evaluate treatment with azithromycin ophthalmic solution (azithromycin). MGD was induced in HR-1 hairless mice by feeding them a special diet with limited lipid content (HR-AD).
    MEHODS. Male HR-1 hairless mice were fed an HR-AD diet for 16 weeks. Development of MGD was assessed by histopathology at 4-week intervals. The lid margin was observed by slit-lamp examination. After cessation of the HR-AD diet, the mice were fed a normal diet to restore normal eye conditions. Expression of cytokeratin 6 was determined by immunostaining. We evaluated the effects of topically applied azithromycin on the plugged orifice in this model.
    RESULTS. After mice were fed the HR-AD diet, histopathology analysis showed hyperkeratinization of the ductal epithelium in the meibomian gland. Ductal hyperkeratinization resulted in the loss of acini, followed by atrophy of the gland. Slit-lamp examination revealed a markedly plugged orifice, telangiectasia, and a toothpaste-like meibum compared with that of a normal eyelid. Cessation of feeding with HR-AD ameliorated both the MGD signs and the expression of cytokeratin 6, restoring the tissue to a histologically normal state. Azithromycin treatment significantly decreased the number of plugged orifices and ameliorated atrophy, as revealed by histopathologic analysis.
    CONCLUSIONS. We developed a novel model that mimics human MGD signs in HR-1 hairless mice fed an HR-AD diet. Azithromycin treatment led to therapeutic improvement in this model. This MGD model could be useful for the evaluation of drug candidates for MGD.

    DOI: 10.1167/iovs.16-19227

    Web of Science

    researchmap

  • Synthesis and in vitro cancer cell growth inhibition evaluation of 11-amino-modified 5-Me-indolo[2,3-b]quinolines and their COMPARE analyses

    Masashi Okada, Zhen-Wu Mei, Md. Imran Hossain, Li Wang, Taihei Tominaga, Takeshi Takebayashi, Masaharu Murakami, Mizuki Yasuda, Tsukasa Shigehiro, Tomonari Kasai, Akifumi Mizutani, Hiroshi Murakami, Ibrahim El Tantawy El Sayed, Shingo Dan, Takao Yamori, Masaharu Seno, Tsutomu Inokuchi

    MEDICINAL CHEMISTRY RESEARCH   25 ( 5 )   879 - 892   2016.5

     More details

    Language:English   Publisher:SPRINGER BIRKHAUSER  

    A plant-derived neocryptolepine core, the 5-Me-indolo[2,3-b]quinoline skeleton, was emblazoned with substituents at C11 and C2 and then tested against various cancer cell lines to find potent anticancer agents. In the in vitro antiproliferative activity assay against the breast cancer MDA-MB-453 cell line, the attachment of alkylamino substituents at C11 of the 5-Me-indolo[2,3-b]quinoline induced improved activities. Specifically, 11-(3-aminopropylamino) and 11-(4-aminobutylamino) derivatives indicated the highest activity and selectivity against MDA-MB-453 (IC50 = 0.3-0.5 mu M) and also exhibited a higher cytotoxicity against the colon adenocarcinoma (WiDr) and ovarian cancer (SKOv3) cell lines. A synergistic effect by attachment of substituents at C2 was favorably observed with an electron-donating group, such as CH3O, and unfavorably observed with an electron-withdrawing one, such as F and CF3. Further modification of the terminal free amino group of the lariat attachment at C11 into the corresponding acylamides and 2,3-dihydrobenzo[e][1,3]thiazin-4-ones was not effective for the antiproliferative activity. The computer-assisted database analysis, COMPARE, suggested that 14e and 13b have a mode of action similar to actinomycin D and 13c has a mode of actions similar to vindesine sulfate or aclarubicin hydrochloride. However, the new compounds may have other unique mode of actions since the correlation coefficients (r) were in relatively low levels.

    DOI: 10.1007/s00044-016-1508-z

    Web of Science

    researchmap

  • Evaluation of glycosylated docetaxel-encapsulated liposomes prepared by remote loading under solubility gradient

    Tsukasa Shigehiro, Wenjia Zhai, Arun Vaidyanath, Junko Masuda, Akifumi Mizutani, Tomonari Kasai, Hiroshi Murakami, Hiroki Hamada, David S. Salomon, Katsuhiko Mikuni, Yuhki Seno, Tadakatsu Mandai, Masaharu Seno

    JOURNAL OF MICROENCAPSULATION   33 ( 2 )   172 - 182   2016.2

     More details

    Language:English   Publisher:TAYLOR & FRANCIS LTD  

    Docetaxel comprises one of the most effective anti-cancer drugs despite of serious side effects. Liposomes encapsulation is practically feasible to deliver the drug. However, due to the significant hydrophobicity, docetaxel will be integrated into the lipid bilayer resulting in poor encapsulation capacity. Here, we evaluated a remote loading strategy using a solubility gradient made between the two solvents for 7-glucosyloxyacetyldocetaxel, which has enhanced water solubility of docetaxel with a coupled glucose moiety. Therefore, 7-glucosyloxyacetyldocetaxel was more effectively encapsulated into liposomes with 71.0% of encapsulation efficiency than docetaxel. While 7-glucosyloxyacetyldocetaxel exhibited 90.9% of tubulin stabilisation activity of docetaxel, 7-glucosyloxyacetyldocetaxel encapsulated in liposomes significantly inhibited the growth of tumour in vivo with side effects less than unencapsulated drug. Collectively, the encapsulation of 7-glucosyloxyacetyldocetaxel into liposomes by remote loading under the solubility gradient is considered to be a promising application to prepare practical drug delivery system.

    DOI: 10.3109/02652048.2016.1144815

    Web of Science

    researchmap

  • 第3節 iPS細胞を用いたがん幹細胞モデル作成のポイントとその評価, 「次世代のがん治療薬・診断のための研究開発~免疫療法・遺伝子治療・がん幹細胞~、第13章 がん幹細胞を標的とした治療薬研究」

    技術情報協会   348 - 353   2016

     More details

  • Release of siRNA from Liposomes Induced by Curcumin

    Kazuyo Fujita, Yoshie Hiramatsu, Hideki Minematsu, Masaharu Somiya, Shun'ichi Kuroda, Masaharu Seno, Shuji Hinuma

    Journal of Nanotechnology   2016   2016

     More details

    Language:English   Publisher:Hindawi Limited  

    Liposomes are a potential carrier of small interfering RNA (siRNA) for drug delivery systems (DDS). In this study, we searched for a molecule capable of controlling the release of siRNA from a certain type of liposomes and found that curcumin could induce the release of siRNA from the liposomes encapsulating siRNA within 30 min. However, the release of siRNA from the liposomes by curcumin showed a unique dose-response (i.e., bell-shaped curve) with a maximal induction at around 60 μg/ml of curcumin. Liposomal lipid compositions and temperatures influenced the efficiency in the release of siRNA induced by curcumin. About 10% of curcumin at a 60 μg/ml dose was incorporated into the liposomes within 30 min under our experimental conditions. Our results suggest a possibility that curcumin is useful in controlling the permeability of liposomes carrying large molecules like siRNA.

    DOI: 10.1155/2016/7051523

    Scopus

    researchmap

  • A Unique Procedure to Identify Cell Surface Markers Through a Spherical Self-Organizing Map Applied to DNA Microarray Analysis.

    Sugii Y, Kasai T, Ikeda M, Vaidyanath A, Kumon K, Mizutani A, Seno A, Tokutaka H, Kudoh T, Seno M

    Biomark Cancer   8   17 - 23   2016

     More details

  • iPSC-derived cancer stem cells provide a model of tumor vasculature

    Marta Prieto-Vila, Ting Yan, Anna Sanchez Calle, Neha Nair, Laura Hurley, Tomonari Kasai, Hiroki Kakuta, Junko Masuda, Hiroshi Murakami, Akifumi Mizutani, Masaharu Seno

    AMERICAN JOURNAL OF CANCER RESEARCH   6 ( 9 )   1906 - 1921   2016

     More details

    Language:English   Publisher:E-CENTURY PUBLISHING CORP  

    To grow beyond a size of approximately 1-2 mm(3), tumor cells activate many processes to develop blood vasculature. Growing evidences indicate that the formation of the tumor vascular network is very complex, and is not restricted to angiogenesis. Cancer cell-derived tumor vasculatures have been recently described. Among them, endothelial differentiation of tumor cells have been directly related to cancer stem cells, which are cells within a tumor that possess the capacity to self-renew, and to exhibit multipotential heterogeneous lineages of cancer cells. Vasculogenic mimicry has been described to be formed by cancer cells expressing stemness markers. Thus, cancer stem cells have been proposed to contribute to vasculogenic mimicry, though its relation is yet to be clarified. Here, we analyzed the tumor vasculature by using a model of mouse cancer stem cells, miPS-LLCcm cells, which we have previously established from mouse induced pluripotent stem cells and we introduced the DsRed gene in miPS-LLCcm to trace them in vivo. Various features of vasculature were evaluated in ovo, in vitro, and in vivo. The tumors formed in allograft nude mice exhibited angiogenesis in chick chorioallantoic membrane assay. In those tumors, along with penetrated host endothelial vessels, we detected endothelial differentiation from cancer stem cells and formation of vasculogenic mimicry. The angiogenic factors such as VEGF-A and FGF2 were expressed predominantly in the cancer stem cells subpopulation of miPS-LLCcm cells. Our results suggested that cancer stem cells play key roles in not only the recruitment of host endothelial vessels into tumor, but also in maturation of endothelial linage of cancer stem cell's progenies. Furthermore, the undifferentiated subpopulation of the miPS-LLCcm participates directly in the vasculogenic mimicry formation. Collectively, we show that miPS-LLCcm cells have advantages to further study tumor vasculature and to develop novel targeting strategies in the future.

    Web of Science

    researchmap

  • A new PDAC mouse model originated from iPSCs-converted pancreatic cancer stem cells (CSCcm)

    Anna Sanchez Calle, Neha Nair, Aung KoKo Oo, Marta Prieto-Vila, Megumi Koga, Apriliana Cahya Khayrani, Maram Hussein, Laura Hurley, Arun Vaidyanath, Akimasa Seno, Yoshiaki Iwasaki, Malu Calle, Tomonari Kasai, Masaharu Seno

    AMERICAN JOURNAL OF CANCER RESEARCH   6 ( 12 )   2799 - 2815   2016

     More details

    Language:English   Publisher:E-CENTURY PUBLISHING CORP  

    Pancreatic ductal adenocarcinoma (PDAC) is the most representative form of pancreatic cancers. PDAC solid tumours are constituted of heterogeneous populations of cells including cancer stem cells (CSCs), differentiated cancer cells, desmoplastic stroma and immune cells. The identification and consequent isolation of pancreatic CSCs facilitated the generation of genetically engineered murine models. Nonetheless, the current models may not be representative for the spontaneous tumour occurrence. In the present study, we show the generation of a novel pancreatic iPSC-converted cancer stem cell lines (CSCcm) as a cutting-edge model for the study of PDAC. The CSCcm lines were achieved only by the influence of pancreatic cancer cell lines conditioned medium and were not subjected to any genetic manipulation. The xenografts tumours from CSCcm lines displayed histopathological features of ADM, PanIN and PDAC lesions. Further molecular characterization from RNA-sequencing analysis highlighted primary culture cell lines (1st CSCcm) as potential candidates to represent the pancreatic CSCs and indicated the establishment of the pancreatic cancer molecular pattern in their subsequent progenies 2nd CSCcm and 3rd CSCcm. In addition, preliminary RNA-seq SNPs analysis showed that the distinct CSCcm lines did not harbour single point mutations for the oncogene Kras codon 12 or 13. Therefore, PDAC-CSCcm model may provide new insights about the actual occurrence of the pancreatic cancer leading to develop different approaches to target CSCs and abrogate the progression of this fatidic disease.

    Web of Science

    researchmap

  • Synthesis, Biological Evaluation, Docking and QSAR Studies of Some Novel Naphthalimide Dithiocarbamate Analogs as Antitumor and Anti-Inflammatory Agents.

    Zahra MH, Osman AMA, Agwa H, Nair N, Calle AS, Hurley L, Farag D, Kasai T, Seno M, Zahran M

    Medicinal Chemistry (Los Angeles)   6 ( 12 )   694 - 703   2016

     More details

  • 球面自己組織化マップを用いたキナーゼパネルアセイデータのクラスタリング

    工藤孝幸, 妹尾昌治

    CCISJ Bulletin   34 ( 1 )   2 - 5   2016

  • がんモデル動物を用いた悪性度の違いによる全身免疫能変化の解析

    増田 潤子, 高山 英次, 佐藤 あやの, 守本 祐司, 本庶 仁子, 石塚 俊晶, 徳野 慎一, 青笹 季文, 光吉 俊二, 重廣 司, 前野 成実, 村上 宏, 笠井 智成, 水谷 昭文, Vaidyanath Arun, 妹尾 彬正, 川木 晴美, 神谷 真子, 野, 近藤 信夫, 一瀬 雅夫, 一戸 辰夫, 妹尾 昌治

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集   88回・38回   [1P1079] - [1P1079]   2015.12

     More details

    Language:Japanese   Publisher:(公社)日本生化学会  

    researchmap

  • Derivation of a model of cancer stem cell from human induced pluripotent stem cells

    Tomonari Kasai, Kenta Hoshikawa, Shuto Takejiri, Masashi Ikeda, Kazuki Kumon, Anna Sanchez Calle, Arun Vaidyanath, Akifumi Mizutani, Chen Ling, Masaharu Seno

    CANCER RESEARCH   75   2015.8

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2015-LB-144

    Web of Science

    researchmap

  • Iron control is a novel therapeutic target of cancer stem cells

    Takayuki Ninomiya, Toshiaki Ohara, Hajime Kashima, Ryoichi Katsube, Kazuhiro Noma, Yasuko Tomono, Akifumi Mizutani, Tomonari Kasai, Masaharu Seno, Shinji Kuroda, Hiroyuki Kishimoto, Hiroshi Tazawa, Yasuhiro Shirakawa, Shunsuke Kagawa, Toshiyoshi Fujiwara

    CANCER RESEARCH   75   2015.8

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2015-4243

    Web of Science

    researchmap

  • Eosinophil Cationic Protein Shows Survival Effect on H9c2 Cardiac Myoblast Cells with Enhanced Phosphorylation of ERK and Akt/GSK-3 beta under Oxidative Stress

    Hiroko Ishii, Shigeshi Kamikawa, Satoshi Hirohata, Akifumi Mizutani, Koji Abe, Masaharu Seno, Toshitaka Ohashi, Yoshifumi Ninomiya

    ACTA MEDICA OKAYAMA   69 ( 3 )   145 - 153   2015.6

     More details

    Language:English   Publisher:OKAYAMA UNIV MED SCHOOL  

    Eosinophil cationic protein (ECP) is well known as a cationic protein contained in the basic granules of activated eosinophils. Recent studies have reported that ECP exhibits novel activities on various types of cells, including rat neonatal cardiomyocytes. Here we evaluated the effects of ECP on rat cardiac myoblast H9c2 cells. Our results showed that ECP enhanced the survival of the cells, in part by promoting the ERK and Akt/GSK-3 beta signaling pathways. ECP attenuated the cytotoxic effects of H2O2 on H9c2 cells as well as the production of reactive oxygen species, the number of apoptotic cells and caspase 3/7 activity in the cells. In conclusion, ECP activated the ERR and Akt/GSK-3 beta pathways, resulting in anti-oxidative effects on H9c2 cells that attenuated apoptosis.

    DOI: 10.18926/AMO/53521

    Web of Science

    researchmap

  • Insight into Cancer Stem Cell Niche; Lessons from Cancer Stem Cell Models Generated In Vitro.

    Biology in Stem Cell Niche, Part of the series Stem Cell Biology and Regenerative Medicine   211 - 226   2015

     More details

  • Spherical Self-Organizing Map Detects MYBL 1 As Candidate Gene for Triple-Negative Breast Cancer.

    Ikeda M, Kumon K, Omoto K, Sugii Y, Mizutani A, Vaidyanath A, Kudoh T, Kasai T, Masuda S, Seno M

    Neurosci Biomed Eng   3 ( 2 )   94 - 101   2015

     More details

  • Bacteria: Prospective Savior in Battle against Cancer

    Neha Nair, Tomonari Kasai, Masaharu Seno

    ANTICANCER RESEARCH   34 ( 11 )   6289 - 6296   2014.11

     More details

    Language:English   Publishing type:Book review, literature introduction, etc.   Publisher:INT INST ANTICANCER RESEARCH  

    Conventional anticancer therapies such as chemotherapy are losing their sheen in the battle against cancer. Therefore, strategies for treatment of cancer need to be constantly modified to fulfill the growing demands of alternative therapies. Several viral and non-viral vectors have been exploited for anticancer gene therapy. But over the years bacteria have been proven to be an important candidate for successful evasion of cancer. They serve as invaluable source of tumor-specific anticancer genes, toxins, polysaccharides for synthesis of nanodrugs and gene-delivery vectors. The current review assesses the role of important bacterial groups in different spheres of anti-cancer research.

    Web of Science

    researchmap

  • Anti-cancer activity of immunoliposomes encapsulated effective amount of glycosylated paclitaxel with novel loading strategy

    Tsukasa Shigehiro, Tomonari Kasai, Akifumi Mizutani, Hiroshi Murakami, Katsuhiko Mikuni, Tadakatsu Mandai, Hiroki Hamada, Masaharu Seno

    CANCER RESEARCH   74 ( 19 )   2014.10

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2014-4461

    Web of Science

    researchmap

  • Efficient Drug Delivery of Paclitaxel Glycoside: A Novel Solubility Gradient Encapsulation into Liposomes Coupled with Immunoliposomes Preparation

    Tsukasa Shigehiro, Tomonari Kasai, Masaharu Murakami, Sreeja C. Sekhar, Yuki Tominaga, Masashi Okada, Takayuki Kudoh, Akifumi Mizutani, Hiroshi Murakami, David S. Salomon, Katsuhiko Mikuni, Tadakatsu Mandai, Hiroki Hamada, Masaharu Seno

    PLOS ONE   9 ( 9 )   e107976   2014.9

     More details

    Language:English   Publisher:PUBLIC LIBRARY SCIENCE  

    Although the encapsulation of paclitaxel into liposomes has been extensively studied, its significant hydrophobic and uncharged character has generated substantial difficulties concerning its efficient encapsulation into the inner water core of liposomes. We found that a more hydrophilic paclitaxel molecule, 7-glucosyloxyacetylpaclitaxel, retained tubulin polymerization stabilization activity. The hydrophilic nature of 7-glucosyloxyacetylpaclitaxel allowed its efficient encapsulation into the inner water core of liposomes, which was successfully accomplished using a remote loading method with a solubility gradient between 40% ethylene glycol and Cremophor EL/ethanol in PBS. Trastuzumab was then conjugated onto the surface of liposomes as immunoliposomes to selectively target human epidermal growth factor receptor-2 (HER2)-overexpressing cancer cells. In vitro cytotoxicity assays revealed that the immunoliposomes enhanced the toxicity of 7-glucosyloxyacetylpaclitaxel in HER2-overexpressing cancer cells and showed more rapid suppression of cell growth. The immunoliposomes strongly inhibited the tumor growth of HT-29 cells xenografted in nude mice. Notably, mice survived when treated with the immunoliposomes formulation, even when administered at a lethal dose of 7glucosyloxyacetylpaclitaxel in vivo. This data successfully demonstrates immunoliposomes as a promising candidate for the efficient delivery of paclitaxel glycoside.

    DOI: 10.1371/journal.pone.0107976

    Web of Science

    researchmap

  • In vitro anti-proliferative and anti-angiogenic activities of thalidomide dithiocarbamate analogs

    Bishoy Y. A. El-Aarag, Tomonari Kasai, Magdy A. H. Zahran, Nadia I. Zakhary, Tsukasa Shigehiro, Sreeja C. Sekhar, Hussein S. Agwa, Akifumi Mizutani, Hiroshi Murakami, Hiroki Kakuta, Masaharu Seno

    INTERNATIONAL IMMUNOPHARMACOLOGY   21 ( 2 )   283 - 292   2014.8

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    Inhibition of angiogenesis is currently perceived as a promising strategy in the treatment of cancer. The anti-angiogenicity of thalidomide has inspired a second wave of research on this teratogenic drug. The present study aimed to investigate the anti-proliferative and anti-angiogenic activities of two thalidomide dithiocarbamate analogs by studying their anti-proliferative effects on human umbilical vein endothelial cells (HUVECs) and MDA-MB-231 human breast cancer cell lines. Their action on the expression levels of IL-6, IL-8, TNF-alpha, VEGF(165), and MMP-2 was also assessed. Furthermore, their effect on angiogenesis was evaluated through wound healing, migration, tube formation, and nitric oxide (NO) assays. Results illustrated that the proliferation of HUVECs and MDA-MB-231 cells was not significantly affected by thalidomide at 625-100 mu M. Thalidomide failed to block angiogenesis at similar concentrations. By contrast, thalidomide dithiocarbamate analogs exhibited significant anti-proliferative action on HUVECs and MDA-MB-231 cells without causing cytotoxicity and also showed powerful anti-angiogenicity in wound healing, migration, tube formation, and NO assays. Thalidomide analogs 1 and 2 demonstrated more potent activity to suppress expression levels of IL-6, IL-8, TNF-alpha, VEGF(165), and MMP-2 than thalidomide. Analog 1 consistently, showed the highest potency and efficacy in all the assays. Taken together, our results support further development and evaluation of novel thalidomide analogs as antitumor and anti-angiogenic agents. (C) 2014 The Authors. Published by Elsevier B.V.

    DOI: 10.1016/j.intimp2014.05.007

    Web of Science

    researchmap

  • Cancer stem cells maintain a hierarchy of differentiation by creating their niche

    Shuichi Matsuda, Ting Yan, Akifumi Mizutani, Tatsuyuki Sota, Yuki Hiramoto, Marta Prieto-Vila, Ling Chen, Ayano Satoh, Takayuki Kudoh, Tomonari Kasai, Hiroshi Murakami, Li Fu, David S. Salomon, Masaharu Seno

    INTERNATIONAL JOURNAL OF CANCER   135 ( 1 )   27 - 36   2014.7

     More details

    Language:English   Publisher:WILEY-BLACKWELL  

    The self-renewal and differentiation properties of cancer stem cells (CSCs) are regulated and maintained by the CSC niche. However, the mechanism of this maintenance, especially the maintenance contributed by differentiated cancer cells, remains to be fully elucidated. Recently, we have established a model of CSCs, miPS-LLCcm, from mouse induced pluripotent stem cells (miPSCs). In vitro cultured miPS-LLCcm cells were autonomously balanced with stem-like cells and differentiated cells including vascular endothelial cells. Under these conditions, the CSC properties appeared to be stable in the presence of the factor(s) secreted by the differentiated cells. The factor(s) activated Notch signaling and promoted self-renewal of CSCs. In addition, the secreted factor(s) appeared to regulate the differentiation lineage of CSCs. Our results indicate that the differentiated progenies of CSCs containing vascular endothelium play important roles for regulating the CSC's properties. Therefore, miPS-LLCcm cells create their own in vitro niche to maintain themselves in the hierarchy of differentiating CSCs.

    DOI: 10.1002/ijc.28648

    Web of Science

    researchmap

  • Cancer stem cells maintain a hierarchy of differentiation by creating their niche

    Shuichi Matsuda, Ting Yan, Akifumi Mizutani, Tatsuyuki Sota, Yuki Hiramoto, Marta Prieto-Vila, Ling Chen, Ayano Satoh, Takayuki Kudoh, Tomonari Kasai, Hiroshi Murakami, Li Fu, David S. Salomon, Masaharu Seno

    INTERNATIONAL JOURNAL OF CANCER   135 ( 1 )   27 - 36   2014.7

     More details

    Language:English   Publisher:WILEY-BLACKWELL  

    The self-renewal and differentiation properties of cancer stem cells (CSCs) are regulated and maintained by the CSC niche. However, the mechanism of this maintenance, especially the maintenance contributed by differentiated cancer cells, remains to be fully elucidated. Recently, we have established a model of CSCs, miPS-LLCcm, from mouse induced pluripotent stem cells (miPSCs). In vitro cultured miPS-LLCcm cells were autonomously balanced with stem-like cells and differentiated cells including vascular endothelial cells. Under these conditions, the CSC properties appeared to be stable in the presence of the factor(s) secreted by the differentiated cells. The factor(s) activated Notch signaling and promoted self-renewal of CSCs. In addition, the secreted factor(s) appeared to regulate the differentiation lineage of CSCs. Our results indicate that the differentiated progenies of CSCs containing vascular endothelium play important roles for regulating the CSC's properties. Therefore, miPS-LLCcm cells create their own in vitro niche to maintain themselves in the hierarchy of differentiating CSCs.

    DOI: 10.1002/ijc.28648

    Web of Science

    researchmap

  • A Cancer Stem Cell Model: An Insight into the Conversion of Induced Pluripotent Stem Cells to Cancer Stem-Like Cells

    Akifumi Mizutani, Ling Chen, Tomonari Kasai, Takayuki Kudoh, Hiroshi Murakami, Li Fu, Masaharu Seno

    Cancer Stem Cells   79 - 87   2014.4

     More details

    Language:English   Publisher:Wiley Blackwell  

    Appropriate model cell lines recapitulating cancer stem cell (CSC) properties would accelerate not only investigation of cancer stem cells but also development of new clinical cancer therapy by establishing a screening system for anti-cancer stem cell agents. In this chapter, the authors introduce their recent work and others to generate cells with cancer stem cell properties in vitro. They also discuss the concept of cancer stem cells with results obtained from originally established cancer stem-like cells. Finally, they describe future applications of the model to basic and clinical studies. In the field of regeneration therapy, the pluripotent stem cells such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are promising sources of differentiated cells for transplantation. Interestingly, these cancer stem-like cells were not required to be introduced with the genes for cell surface markers that are commonly used to characterize and isolate cancer stem cells.

    DOI: 10.1002/9781118356203.ch6

    Scopus

    researchmap

  • Development of In-111-Labeled Liposomes for Vulnerable Atherosclerotic Plaque Imaging

    Mikako Ogawa, Izumi O. Umeda, Mutsumi Kosugi, Ayumi Kawai, Yuka Hamaya, Misato Takashima, Hongxia Yin, Takayuki Kudoh, Masaharu Seno, Yasuhiro Magata

    JOURNAL OF NUCLEAR MEDICINE   55 ( 1 )   115 - 120   2014.1

     More details

    Language:English   Publisher:SOC NUCLEAR MEDICINE INC  

    Macrophage infiltration is a common characteristic feature of atherosclerotic-vulnerable plaques. Macrophages recognize phosphatidylserine (PS) exposed on the surface of apoptotic cells, which triggers the engulfment of the apoptotic cells by macrophages through phagocytosis. In this study, we prepared radiolabeled PS liposomes for detection of vulnerable plaques. Methods: PS liposomes were prepared by lipid film hydration. Phosphatidylcholine (PC) liposomes were prepared as controls. Liposomes (100 or 200 nm) were generated by an extruder to produce PS100, PS200, PC100, and PC200 liposomes. These were then radiolabeled by encapsulating In-111-nitrilotriacetic acid using an active-loading method. In-111 liposomes were incubated with cultured macrophages for 2 h, and the uptake level was measured. For biodistribution studies, the In-111 liposomes were injected intravenously into ddY mice. In addition, the In-111 liposomes were injected into apolipoprotein E-deficient (apoE-/-) mice, and the aortas were harvested for autoradiography and oil red O staining. For SPECT imaging, In-111 liposomes were injected intravenously into Watanabe heritable hyperlipidemic rabbits and scanned 48 h after injection. Results: The radiochemical yields were greater than 95% for all the prepared In-111 liposomes. The level of in vitro uptake by macrophages was 60.5, 14.7, 32.0, and 14.4 percentage injected dose per milligram of protein for In-111-PS100, In-111-PC100, In-111-PS200, and In-111-PC200, respectively. In biodistribution studies, high spleen uptake was seen with PC liposomes. Liver uptake was high for all liposomes but was lowest with In-111-PS200. The blood half-lives were 3.2, 22.0, 3.6, and 7.4 min for In-111-PS100, In-111-PC100, In-111-PS200, and In-111-PC200, respectively. The distribution of In-111-labeled PS liposomes into atherosclerotic regions determined by autoradiography was well matched with the results of oil red O staining in apoE-/- mice. The target-to-nontarget ratios were 2.62, 2.23, 3.27, and 2.51 for In-111-PS100, In-111-PC100, In-111-PS200, and In-111-PC200, respectively. The aorta was successfully visualized by SPECT at 48 h after In-111-labeled PS liposome injection; however, high liver uptake was also observed. Discussion: From the in vitro uptake study, it has been demonstrated that macrophage targeting was accomplished by PS modification. Also, an atherosclerotic region was successfully detected by In-111-PS200 in apoE-/- mice and Watanabe heritable hyperlipidemic rabbits in vivo. Liposome modification to obtain slower blood clearance and lower liver uptake would be required to improve the SPECT images.

    DOI: 10.2967/jnumed.113.123158

    Web of Science

    researchmap

  • Chlorotoxin-Fc Fusion Inhibits Release of MMP-2 from Pancreatic Cancer Cells

    Samah El-Ghlban, Tomonari Kasai, Tsukasa Shigehiro, Hong Xia Yin, Sreeja Sekhar, Mikiko Ida, Anna Sanchez, Akifumi Mizutani, Takayuki Kudoh, Hiroshi Murakami, Masaharu Seno

    BIOMED RESEARCH INTERNATIONAL   152659   2014

     More details

    Language:English   Publisher:HINDAWI PUBLISHING CORPORATION  

    Chlorotoxin (CTX) is a 36-amino acid peptide derived from Leiurus quinquestriatus (scorpion) venom, which inhibits low-conductance chloride channels in colonic epithelial cells. It has been reported that CTX also binds to matrix metalloproteinase-2 (MMP-2), membrane type-1 MMP, and tissue inhibitor of metalloproteinase-2, as well as CLC-3 chloride ion channels and other proteins. Pancreatic cancer cells require the activation of MMP-2 during invasion and migration. In this study, the fusion protein was generated by joining the CTX peptide to the amino terminus of the human IgG-Fc domain without a hinge domain, the monomeric form of chlorotoxin (M-CTX-Fc). The resulting fusion protein was then used to target pancreatic cancer cells (PANC-1) in vitro. M-CTX-Fc decreased MMP-2 release into the media of PANC-1 cells in a dose-dependent manner. M-CTX-Fc internalization into PANC-1 cells was observed. When the cells were treated with chlorpromazine (CPZ), the internalization of the fusion protein was reduced, implicating a clathrin-dependent internalization mechanism of M-CTX-Fc in PANC-1 cells. Furthermore, M-CTX-Fc clearly exhibited the inhibition of the migration depending on the concentration, but human IgG, as negative control of Fc, was not affected. The M-CTX-Fc may be an effective instrument for targeting pancreatic cancer.

    DOI: 10.1155/2014/152659

    Web of Science

    researchmap

  • Mutual dependence between cancer stem cells and their progenies: the niche created by the progenies is sustaining cancer stem cells.

    Cancer Cell & Microenvironment   1 ( 4 )   141 - 144   2014

     More details

  • Cancer stem cells converted from pluripotent stem cells and the cancerous niche.

    Kasai T, Chen L, Mizutani A, Kudoh T, Murakami H, Fu L, Seno M

    Journal of Stem cells & regenerative medicine   10 ( 1 )   2 - 7   2014

     More details

  • Characterization of Cancer Stem-Like Cells Derived from Mouse Induced Pluripotent Stem Cells Transformed by Tumor-Derived Extracellular Vesicles

    Ting Yan, Akifumi Mizutani, Ling Chen, Mai Takaki, Yuki Hiramoto, Shuichi Matsuda, Tsukasa Shigehiro, Tomonari Kasai, Takayuki Kudoh, Hiroshi Murakami, Junko Masuda, Mary J. C. Hendrix, Luigi Strizzi, David S. Salomon, Li Fu, Masaharu Seno

    JOURNAL OF CANCER   5 ( 7 )   572 - 584   2014

     More details

    Language:English   Publisher:IVYSPRING INT PUBL  

    Several studies have shown that cancer niche can perform an active role in the regulation of tumor cell maintenance and progression through extracellular vesicles-based intercellular communication. However, it has not been reported whether this vesicle-mediated communication affects the malignant transformation of normal stem cells/progenitors. We have previously reported that the conditioned medium derived from the mouse Lewis Lung Carcinoma (LLC) cell line can convert mouse induced pluripotent stem cells (miPSCs) into cancer stem cells (CSCs), indicating that normal stem cells when placed in an aberrant microenvironment can give rise to functionally active CSCs. Here, we focused on the contribution of tumor-derived extracellular vesicles (tEVs) that are secreted from LLC cells to induce the transformation of miPSCs into CSCs. We isolated tEVs from the conditioned medium of LLC cells, and then the differentiating miPSCs were exposed to tEVs for 4 weeks. The resultant tEV treated cells (miPS-LLCev) expressed Nanog and Oct3/4 proteins comparable to miPSCs. The frequency of sphere formation of the miPS-LLCev cells in suspension culture indicated that the self-renewal capacity of the miPS-LLCev cells was significant. When the miPS-LLCev cells were subcutaneously transplanted into Balb/c nude mice, malignant liposarcomas with extensive angiogenesis developed. miPS-LLCevPT and miPS-LLCevDT, the cells established from primary site and disseminated liposarcomas, respectively, showed their capacities to self-renew and differentiate into adipocytes and endothelial cells. Moreover, we confirmed the secondary liposarcoma development when these cells were transplanted. Taken together, these results indicate that miPS-LLCev cells possess CSC properties. Thus, our current study provides the first evidence that tEVs have the potential to induce CSC properties in normal tissue stem cells/progenitors.

    DOI: 10.7150/jca.8865

    Web of Science

    researchmap

  • Mouse induced pluripotent stem cell microenvironment generates epithelial-mesenchymal transition in mouse Lewis lung cancer cells

    Ling Chen, Akifumi Mizutani, Tomonari Kasai, Ting Yan, Guoliang Jin, Arun Vaidyanath, Bishoy Y. A. El-Aarag, Yixin Liu, Takayuki Kudoh, David S. Salomon, Li Fu, Masaharu Seno

    AMERICAN JOURNAL OF CANCER RESEARCH   4 ( 1 )   80 - U92   2014

     More details

    Language:English   Publisher:E-CENTURY PUBLISHING CORP  

    Induced pluripotent stem (iPS) cells may be a powerful tool in regenerative medicine, but their potential tumorigenicity is a significant challenge for the clinical use of iPS cells. Previously, we succeeded in converting miPS cells into cancer stem cells (CSCs) under the conditions of tumor microenvironment. Both stem cells and tumor cells are profoundly influenced by bi-directional communication with their respective microenvironment, which dictates cell fate determination and behavior. The microenvironment derived from iPS cells has not been well studied. In this paper, we have investigated the effects of secreted factors from Nanog-mouse iPS (miPS) cells on mouse Lewis lung cancer (LLC) cells that are found in the conditioned media. The results demonstrated that miPS cells secrete factors that can convert the epithelia phenotype of LLC cells to a mesenchymal phenotype, and that can promote tumorigenisity, migration and invasion. Furthermore, LLC cells that have been exposed to miPS conditioned medium became resistant to apoptosis. These various biological effects suggest that the miPS microenvironment contain factors that can promote an epithelial-mesenchymal transition (EMT) through an active Snail-MMP axis or by suppressing differentiation in LLC cells.

    Web of Science

    researchmap

  • High epiregulin expression in human U87 glioma cells relies on IRE1 alpha and promotes autocrine growth through EGF receptor

    Gregor Auf, Arnaud Jabouille, Maylis Delugin, Sylvaine Guerit, Raphael Pineau, Sophie North, Natalia Platonova, Marlene Maitre, Alexandre Favereaux, Peter Vajkoczy, Masaharu Seno, Andreas Bikfalvi, Dmitri Minchenko, Oleksandr Minchenko, Michel Moenner

    BMC CANCER   13   13 - 597   2013.12

     More details

    Language:English   Publisher:BIOMED CENTRAL LTD  

    Background: Epidermal growth factor (EGF) receptors contribute to the development of malignant glioma. Here we considered the possible implication of the EGFR ligand epiregulin (EREG) in glioma development in relation to the activity of the unfolded protein response (UPR) sensor IRE1 alpha. We also examined EREG status in several glioblastoma cell lines and in malignant glioma.
    Methods: Expression and biological properties of EREG were analyzed in human glioma cells in vitro and in human tumor xenografts with regard to the presence of ErbB proteins and to the blockade of IRE1 alpha. Inactivation of IRE1 alpha was achieved by using either the dominant-negative strategy or siRNA-mediated knockdown.
    Results: EREG was secreted in high amounts by U87 cells, which also expressed its cognate EGF receptor (ErbB1). A stimulatory autocrine loop mediated by EREG was evidenced by the decrease in cell proliferation using specific blocking antibodies directed against either ErbB1 (cetuximab) or EREG itself. In comparison, anti-ErbB2 antibodies (trastuzumab) had no significant effect. Inhibition of IRE1 alpha dramatically reduced EREG expression both in cell culture and in human xenograft tumor models. The high-expression rate of EREG in U87 cells was therefore linked to IRE1 alpha, although being modestly affected by chemical inducers of the endoplasmic reticulum stress. In addition, IRE1-mediated production of EREG did not depend on IRE1 RNase domain, as neither the selective dominant-negative invalidation of the RNase activity (IRE1 kinase active) nor the siRNA-mediated knockdown of XBP1 had significant effect on EREG expression. Finally, chemical inhibition of c-Jun N-terminal kinases (JNK) using the SP600125 compound reduced the ability of cells to express EREG, demonstrating a link between the growth factor production and JNK activation under the dependence of IRE1 alpha.
    Conclusion: EREG may contribute to glioma progression under the control of IRE1 alpha, as exemplified here by the autocrine proliferation loop mediated in U87 cells by the growth factor through ErbB1.

    DOI: 10.1186/1471-2407-13-597

    Web of Science

    researchmap

  • Cytokine expression in human dermal fibroblasts stimulated with eosinophil cationic protein measured by protein array

    Takamaro Sato, Yoshihiko Soga, Tomoko Yamaguchi, Michio Meguro, Hiroshi Maeda, Joji Tada, Takayuki Otani, Masaharu Seno, Shogo Takashiba

    ASIAN PACIFIC JOURNAL OF ALLERGY AND IMMUNOLOGY   31 ( 4 )   271 - 276   2013.12

     More details

    Language:English   Publisher:ALLERGY IMMUNOL SOC THAILAND,  

    Background: Eosinophil cationic protein (ECP) was reported previously to be involved in allergic inflammation with cytotoxic activity. On the other hand, recent studies showed that ECP did not induce cell death but inhibited the growth of cancer-derived cells. Our previous study indicated that human ECP enhanced differentiation of rat neonatal cardiomyocytes and stress fiber formation in Balb/c 3T3 mouse fibroblasts, while the effects of human ECP on human fibroblasts are unknown.
    Objective: The present study was performed to determine the effects of human ECP on cytokine expression in human fibroblasts by protein array.
    Methods: The effects of recombinant human ECP (rhECP) on normal human dermal fibroblasts (NHDF) were examined by assaying cell growth. Furthermore, cytokine expression of NHDF stimulated by ECP, which could influence cell growth, was evaluated by protein array.
    Results: ECP was not cytotoxic but enhanced the growth of NHDF. The peak rhECP concentration that enhanced the cell counts by 1.56-fold was 100 ng/mL, which was significantly different from cultures without ECP stimulation (ANOVA/ Scheffe's test, P &lt; 0.05). Array analyses indicated that ciliary neurotrophic factor (CNTF), neutrophil-activating peptide (NAP)-2, and neurotrophin (NT)-3 were significantly upregulated in NHDF stimulated with 100 ng/mL ECP compared to those without stimulation.
    Conclusion: ECP is not cytotoxic but enhances the growth of NHDF. CNTF, NAP-2, and NT-3 were suggested to be involved in enhancing the growth of NHDF. These findings will contribute to determination of the role of ECP in allergic inflammation.

    DOI: 10.12932/AP0287.31.4.2013

    Web of Science

    researchmap

  • Steering Target Selectivity and Potency by Fragment-Based De Novo Drug Design

    Tiago Rodrigues, Takayuki Kudoh, Filip Roudnicky, Yi Fan Lim, Yen-Chu Lin, Christian P. Koch, Masaharu Seno, Michael Detmar, Gisbert Schneider

    ANGEWANDTE CHEMIE-INTERNATIONAL EDITION   52 ( 38 )   10006 - 10009   2013.9

     More details

    Language:English   Publisher:WILEY-V C H VERLAG GMBH  

    DOI: 10.1002/anie.201304847

    Web of Science

    researchmap

  • Steering Target Selectivity and Potency by Fragment-Based De Novo Drug Design

    Tiago Rodrigues, Takayuki Kudoh, Filip Roudnicky, Yi Fan Lim, Yen-Chu Lin, Christian P. Koch, Masaharu Seno, Michael Detmar, Gisbert Schneider

    ANGEWANDTE CHEMIE-INTERNATIONAL EDITION   52 ( 38 )   10006 - 10009   2013.9

     More details

    Language:English   Publisher:WILEY-V C H VERLAG GMBH  

    DOI: 10.1002/anie.201304847

    Web of Science

    researchmap

  • Nano-micrometer-architectural acidic silica prepared from iron oxide of leptothrix ochracea origin

    Hideki Hashimoto, Atsushi Itadani, Takayuki Kudoh, Satoshi Fukui, Yasushige Kuroda, Masaharu Seno, Yoshihiro Kusano, Yasunori Ikeda, Yasuhiko Benino, Tokuro Nanba, Makoto Nakanishi, Tatsuo Fujii, Jun Takada

    ACS Applied Materials and Interfaces   5 ( 11 )   5194 - 5200   2013.6

     More details

    Language:English  

    We prepared nano-micrometer-architectural acidic silica from a natural amorphous iron oxide with structural silicon which is a product of the iron-oxidizing bacterium Leptothrix ochracea. The starting material was heat-treated at 500 C in a H2 gas flow leading to segregation of α-Fe crystalline particles and then dissolved in 1 M hydrochloric acid to remove the α-Fe particles, giving a gray-colored precipitate. It was determined to be amorphous silica containing some amount of iron (Si/Fe = ∼60). The amorphous silica maintains the nano-microstructure of the starting material - ∼1-μm-diameter micrometer-tubules consisting of inner globular and outer fibrillar structures several tens of nanometer in size - and has many large pores which are most probably formed as a result of segregation of the α-Fe particles on the micrometer-tubule wall. The smallest particle size of the amorphous silica is ∼10 nm, and it has a large surface area of 550 m2/g with micropores (0.7 nm). By using pyridine vapor as a probe molecule to evaluate the active sites in the amorphous silica, we found that it has relatively strong Brønsted and Lewis acidic centers that do not desorb pyridine, even upon evacuation at 400 C. The acidity of this new silica material was confirmed through representative two catalytic reactions: ring-opening reaction and Friedel-Crafts-type reaction, both of which are known to require acid catalysts. © 2013 American Chemical Society.

    DOI: 10.1021/am401029r

    Scopus

    PubMed

    researchmap

  • Nano-micrometer-architectural acidic silica prepared from iron oxide of leptothrix ochracea origin

    Hideki Hashimoto, Atsushi Itadani, Takayuki Kudoh, Satoshi Fukui, Yasushige Kuroda, Masaharu Seno, Yoshihiro Kusano, Yasunori Ikeda, Yasuhiko Benino, Tokuro Nanba, Makoto Nakanishi, Tatsuo Fujii, Jun Takada

    ACS Applied Materials and Interfaces   5 ( 11 )   5194 - 5200   2013.6

     More details

    Language:English  

    We prepared nano-micrometer-architectural acidic silica from a natural amorphous iron oxide with structural silicon which is a product of the iron-oxidizing bacterium Leptothrix ochracea. The starting material was heat-treated at 500 C in a H2 gas flow leading to segregation of α-Fe crystalline particles and then dissolved in 1 M hydrochloric acid to remove the α-Fe particles, giving a gray-colored precipitate. It was determined to be amorphous silica containing some amount of iron (Si/Fe = ∼60). The amorphous silica maintains the nano-microstructure of the starting material - ∼1-μm-diameter micrometer-tubules consisting of inner globular and outer fibrillar structures several tens of nanometer in size - and has many large pores which are most probably formed as a result of segregation of the α-Fe particles on the micrometer-tubule wall. The smallest particle size of the amorphous silica is ∼10 nm, and it has a large surface area of 550 m2/g with micropores (0.7 nm). By using pyridine vapor as a probe molecule to evaluate the active sites in the amorphous silica, we found that it has relatively strong Brønsted and Lewis acidic centers that do not desorb pyridine, even upon evacuation at 400 C. The acidity of this new silica material was confirmed through representative two catalytic reactions: ring-opening reaction and Friedel-Crafts-type reaction, both of which are known to require acid catalysts. © 2013 American Chemical Society.

    DOI: 10.1021/am401029r

    Scopus

    PubMed

    researchmap

  • Eosinophil cationic protein enhances stabilization of beta-catenin during cardiomyocyte differentiation in P19CL6 embryonal carcinoma cells

    Guoliang Jin, Akifumi Mizutani, Takayuki Fukuda, Takayuki Otani, Ting Yan, Marta Prieto Vila, Hiroshi Murakami, Takayuki Kudoh, Satoshi Hirohata, Tomonari Kasai, David S. Salomon, Masaharu Seno

    MOLECULAR BIOLOGY REPORTS   40 ( 4 )   3165 - 3171   2013.4

     More details

    Language:English   Publisher:SPRINGER  

    Prior to gastrulation, the Wnt signaling pathway through stabilized beta-catenin enhances the differentiation of mouse ES cell into cardiomyocytes. We have recently shown that cardiomyocyte differentiation is enhanced by eosinophil cationic protein (ECP) through accelerated expression of marker genes of early cardiac differentiation. Furthermore, ECP enhanced the expression of Wnt3a in P19CL6 cells which were stimulated to differentiate into cardiomyocytes by DMSO. Following these findings, we evaluated in this study the potential of ECP to activate the Wnt/beta-catenin signaling pathway during cardiomyocyte differentiation. Analysis by real time qPCR revealed that ECP increased the expression of Frizzled genes such as Frizzled-1, -2, -4 and -10 in P19CL6 cells in the presence of DMSO. The increased expression of those Wnt receptors was found to inhibit the phosphorylation of beta-catenin resulting in the stabilization and translocation of beta-catenin into the nucleus of P19CL6 cells during the early stages of cardiomyocyte differentiation. When assessed for beta-catenin/TCF transcriptional activity with a TCF-luciferase (TOP/FOP) assay, ECP enhanced luciferase activity in P19CL6 cells during 48 h after transfection with TOP/FOP flash reporter in a stoichiometric manner. Collectively, this suggests that ECP can activate a canonical Wnt/beta-catenin signaling pathway by enhancing the stabilization of beta-catenin during cardiomyocyte differentiation.

    DOI: 10.1007/s11033-012-2390-5

    Web of Science

    researchmap

  • Eosinophil cationic protein enhances stabilization of beta-catenin during cardiomyocyte differentiation in P19CL6 embryonal carcinoma cells

    Guoliang Jin, Akifumi Mizutani, Takayuki Fukuda, Takayuki Otani, Ting Yan, Marta Prieto Vila, Hiroshi Murakami, Takayuki Kudoh, Satoshi Hirohata, Tomonari Kasai, David S. Salomon, Masaharu Seno

    MOLECULAR BIOLOGY REPORTS   40 ( 4 )   3165 - 3171   2013.4

     More details

    Language:English   Publisher:SPRINGER  

    Prior to gastrulation, the Wnt signaling pathway through stabilized beta-catenin enhances the differentiation of mouse ES cell into cardiomyocytes. We have recently shown that cardiomyocyte differentiation is enhanced by eosinophil cationic protein (ECP) through accelerated expression of marker genes of early cardiac differentiation. Furthermore, ECP enhanced the expression of Wnt3a in P19CL6 cells which were stimulated to differentiate into cardiomyocytes by DMSO. Following these findings, we evaluated in this study the potential of ECP to activate the Wnt/beta-catenin signaling pathway during cardiomyocyte differentiation. Analysis by real time qPCR revealed that ECP increased the expression of Frizzled genes such as Frizzled-1, -2, -4 and -10 in P19CL6 cells in the presence of DMSO. The increased expression of those Wnt receptors was found to inhibit the phosphorylation of beta-catenin resulting in the stabilization and translocation of beta-catenin into the nucleus of P19CL6 cells during the early stages of cardiomyocyte differentiation. When assessed for beta-catenin/TCF transcriptional activity with a TCF-luciferase (TOP/FOP) assay, ECP enhanced luciferase activity in P19CL6 cells during 48 h after transfection with TOP/FOP flash reporter in a stoichiometric manner. Collectively, this suggests that ECP can activate a canonical Wnt/beta-catenin signaling pathway by enhancing the stabilization of beta-catenin during cardiomyocyte differentiation.

    DOI: 10.1007/s11033-012-2390-5

    Web of Science

    researchmap

  • A novel remote loading method with solubility gradient to encapsulate effevtive amount of taxanes into liposomes.

    Tsukasa Shigehiro, Tomonari Kasai, Akifumi Mizutani, Hiroshi Murakami, Katsuhiko Mikuni, Tadakatsu Mandai, Hiroki Hamada, Masaharu Seno

    CANCER RESEARCH   73 ( 8 )   2013.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2013-LB-8

    Web of Science

    researchmap

  • Acidic Amorphous Silica Prepared from Iron Oxide of Bacterial Origin

    Hideki Hashimoto, Atsushi Itadani, Takayuki Kudoh, Yasushige Kuroda, Masaharu Seno, Yoshihiro Kusano, Yasunori Ikeda, Makoto Nakanishi, Tatsuo Fujii, Jun Takada

    ACS APPLIED MATERIALS & INTERFACES   5 ( 3 )   518 - 523   2013.2

     More details

    Language:English   Publisher:AMER CHEMICAL SOC  

    Microporous and mesoporous silica derived from biogenous iron oxide is an attractive catalyst for various organic reactions. Biogenous iron oxide contains structural silicon, and amorphous silica remains after iron oxide is dissolved in concentrated hydrochloric acid. The amorphous silica containing slight amounts of iron (Si/Fe = similar to 150) is composed of similar to 6-nm-diameter granular particles. The amorphous silica has a large surface area of 540 m(2)/g with micropores (1.4 nm) and mesopores (&lt;3 nm). By using pyridine vapor as a probe molecule to evaluate the active sites in the amorphous silica, it was found that this material has strong Bronsted and Lewis acid sites. When the catalytic performance of this material was evaluated for reactions including the ring opening of epoxides and Friedel Crafts-type alkylations, which are known to be catalyzed by acid catalysts, this material showed yields higher than those obtained with common silica materials.

    DOI: 10.1021/am302837p

    Web of Science

    researchmap

  • Acidic Amorphous Silica Prepared from Iron Oxide of Bacterial Origin

    Hideki Hashimoto, Atsushi Itadani, Takayuki Kudoh, Yasushige Kuroda, Masaharu Seno, Yoshihiro Kusano, Yasunori Ikeda, Makoto Nakanishi, Tatsuo Fujii, Jun Takada

    ACS APPLIED MATERIALS & INTERFACES   5 ( 3 )   518 - 523   2013.2

     More details

    Language:English   Publisher:AMER CHEMICAL SOC  

    Microporous and mesoporous silica derived from biogenous iron oxide is an attractive catalyst for various organic reactions. Biogenous iron oxide contains structural silicon, and amorphous silica remains after iron oxide is dissolved in concentrated hydrochloric acid. The amorphous silica containing slight amounts of iron (Si/Fe = similar to 150) is composed of similar to 6-nm-diameter granular particles. The amorphous silica has a large surface area of 540 m(2)/g with micropores (1.4 nm) and mesopores (&lt;3 nm). By using pyridine vapor as a probe molecule to evaluate the active sites in the amorphous silica, it was found that this material has strong Bronsted and Lewis acid sites. When the catalytic performance of this material was evaluated for reactions including the ring opening of epoxides and Friedel Crafts-type alkylations, which are known to be catalyzed by acid catalysts, this material showed yields higher than those obtained with common silica materials.

    DOI: 10.1021/am302837p

    Web of Science

    researchmap

  • Cripto-1 enhances the canonical Wnt/β-catenin signaling pathway by binding to LRP5 and LRP6 co-receptors

    Tadahiro Nagaoka, Hideaki Karasawa, Thomas Turbyville, Maria-Cristina Rangel, Nadia P. Castro, Monica Gonzales, Alyson Baker, Masaharu Seno, Stephen Lockett, Yoshimi E. Greer, Jeffrey S. Rubin, David S. Salomon, Caterina Bianco

    Cellular Signalling   25 ( 1 )   178 - 189   2013.1

     More details

    Language:English  

    Cripto-1 is implicated in multiple cellular events, including cell proliferation, motility and angiogenesis, through the activation of an intricate network of signaling pathways. A crosstalk between Cripto-1 and the canonical Wnt/β-catenin signaling pathway has been previously described. In fact, Cripto-1 is a downstream target gene of the canonical Wnt/β-catenin signaling pathway in the embryo and in colon cancer cells and T-cell factor (Tcf)/lymphoid enhancer factor binding sites have been identified in the promoter and the first intronic region of the mouse and human Cripto-1 genes. We now demonstrate that Cripto-1 modulates signaling through the canonical Wnt/β-catenin/Tcf pathway by binding to the Wnt co-receptors low-density lipoprotein receptor-related protein (LRP) 5 and LRP6, which facilitates Wnt3a binding to LRP5 and LRP6. Cripto-1 functionally enhances Wnt3a signaling through cytoplasmic stabilization of β-catenin and elevated β-catenin/Tcf transcriptional activation. Conversely, Wnt3a further increases Cripto-1 stimulation of migration, invasion and colony formation in soft agar of HC11 mouse mammary epithelial cells, indicating that Cripto-1 and the canonical Wnt/β-catenin signaling co-operate in regulating motility and in vitro transformation of mammary epithelial cells. © 2012.

    DOI: 10.1016/j.cellsig.2012.09.024

    Scopus

    PubMed

    researchmap

  • Cripto-1 enhances the canonical Wnt/β-catenin signaling pathway by binding to LRP5 and LRP6 co-receptors

    Tadahiro Nagaoka, Hideaki Karasawa, Thomas Turbyville, Maria-Cristina Rangel, Nadia P. Castro, Monica Gonzales, Alyson Baker, Masaharu Seno, Stephen Lockett, Yoshimi E. Greer, Jeffrey S. Rubin, David S. Salomon, Caterina Bianco

    Cellular Signalling   25 ( 1 )   178 - 189   2013.1

     More details

    Language:English  

    Cripto-1 is implicated in multiple cellular events, including cell proliferation, motility and angiogenesis, through the activation of an intricate network of signaling pathways. A crosstalk between Cripto-1 and the canonical Wnt/β-catenin signaling pathway has been previously described. In fact, Cripto-1 is a downstream target gene of the canonical Wnt/β-catenin signaling pathway in the embryo and in colon cancer cells and T-cell factor (Tcf)/lymphoid enhancer factor binding sites have been identified in the promoter and the first intronic region of the mouse and human Cripto-1 genes. We now demonstrate that Cripto-1 modulates signaling through the canonical Wnt/β-catenin/Tcf pathway by binding to the Wnt co-receptors low-density lipoprotein receptor-related protein (LRP) 5 and LRP6, which facilitates Wnt3a binding to LRP5 and LRP6. Cripto-1 functionally enhances Wnt3a signaling through cytoplasmic stabilization of β-catenin and elevated β-catenin/Tcf transcriptional activation. Conversely, Wnt3a further increases Cripto-1 stimulation of migration, invasion and colony formation in soft agar of HC11 mouse mammary epithelial cells, indicating that Cripto-1 and the canonical Wnt/β-catenin signaling co-operate in regulating motility and in vitro transformation of mammary epithelial cells. © 2012.

    DOI: 10.1016/j.cellsig.2012.09.024

    Scopus

    PubMed

    researchmap

  • マウスiPS細胞から作るがん幹細胞モデル

    笠井智成, 陳 凌, 工藤孝幸, 水谷昭文, 妹尾昌治

    細胞工学   32 ( 3 )   330 - 337   2013

     More details

  • Theranostic protein targeting ErbB2 for bioluminescence imaging and therapy for cancer. International journal

    Xiao-Jian Han, Ling-Fei Sun, Yuki Nishiyama, Bin Feng, Hiroyuki Michiue, Masaharu Seno, Hideki Matsui, Kazuhito Tomizawa

    PloS one   8 ( 9 )   e75288 - e75288   2013

     More details

    Language:English  

    A combination of molecular-targeted cancer imaging and therapy is an emerging strategy to improve cancer diagnosis and minimize the side effects of conventional treatments. Here, we generated a recombinant protein, EC1-GLuc-p53C, by fusing EC1 peptide, an artificial ligand of ErbB2, with Gaussia luciferase (GLuc) and a p53-activating peptide, p53C. EC1-GLuc-p53C was expressed and purified from E. coli BL21. In vitro experiments showed that EC1-GLuc-p53c was stable in luminescent activity and selectively targeted ErbB2-overexpressing BT474 cells for bioluminescence imaging. Moreover, the internalized EC1-GLuc-p53C in BT474 cells exerted its function to reactivate p53 and significantly inhibited cellular proliferation. In tumor-bearing mice, the ErbB2-targeted bioluminescence imaging and therapeutic effect of EC1-GLuc-p53C were also observed specifically in BT474 tumors but not in MCF7 tumors, which does not overexpress ErbB2. Thus, the present study demonstrates EC1-GLuc-p53C to be an effective theranostic reagent targeting ErbB2 for bioluminescence imaging and cancer therapy.

    DOI: 10.1371/journal.pone.0075288

    PubMed

    researchmap

  • Theranostic protein targeting ErbB2 for bioluminescence imaging and therapy for cancer. International journal

    Xiao-Jian Han, Ling-Fei Sun, Yuki Nishiyama, Bin Feng, Hiroyuki Michiue, Masaharu Seno, Hideki Matsui, Kazuhito Tomizawa

    PloS one   8 ( 9 )   e75288   2013

     More details

    Language:English  

    A combination of molecular-targeted cancer imaging and therapy is an emerging strategy to improve cancer diagnosis and minimize the side effects of conventional treatments. Here, we generated a recombinant protein, EC1-GLuc-p53C, by fusing EC1 peptide, an artificial ligand of ErbB2, with Gaussia luciferase (GLuc) and a p53-activating peptide, p53C. EC1-GLuc-p53C was expressed and purified from E. coli BL21. In vitro experiments showed that EC1-GLuc-p53c was stable in luminescent activity and selectively targeted ErbB2-overexpressing BT474 cells for bioluminescence imaging. Moreover, the internalized EC1-GLuc-p53C in BT474 cells exerted its function to reactivate p53 and significantly inhibited cellular proliferation. In tumor-bearing mice, the ErbB2-targeted bioluminescence imaging and therapeutic effect of EC1-GLuc-p53C were also observed specifically in BT474 tumors but not in MCF7 tumors, which does not overexpress ErbB2. Thus, the present study demonstrates EC1-GLuc-p53C to be an effective theranostic reagent targeting ErbB2 for bioluminescence imaging and cancer therapy.

    DOI: 10.1371/journal.pone.0075288

    PubMed

    researchmap

  • Identification of Caveolin-1 as a Potential Causative Factor in the Generation of Trastuzumab Resistance in Breast Cancer Cells

    Sreeja C. Sekhar, Tomonari Kasai, Ayano Satoh, Tsukasa Shigehiro, Akifumi Mizutani, Hiroshi Murakami, Bishoy Y. A. El-Aarag, David S. Salomon, Anna Massaguer, Rafael de Llorens, Masaharu Seno

    JOURNAL OF CANCER   4 ( 5 )   391 - 401   2013

     More details

    Language:English   Publisher:IVYSPRING INT PUBL  

    The oncogenic tyrosine kinase receptor ErbB2 is a prognostic factor and target for breast cancer therapeutics. In contrast with the other ErbB receptors, ErbB2 is hardly internalized by ligand induced mechanisms, indicating a prevalent surface expression. Elevated levels of ErbB2 in tumor cells are associated with its defective endocytosis and down regulation. Here we show that caveolin-1 expression in breast cancer derived SKBR-3 cells (SKBR-3/Cav-1) facilitates ligand induced ErbB2 endocytosis using an artificial peptide ligand EC-eGFP. Similarly, stimulation with humanized anti ErbB2 antibody Trastuzumab (Herceptin) was found to be internalized and co-localized with caveolin-1 in SKBR-3/Cav-1 cells. Internalized EC-eGFP and Trastuzumab in SKBR-3/Cav-1 cells were then delivered via caveolae to the caveolin-1 containing early endosomes. Consequently, attenuated Fc receptor mediated ADCC functions were observed when exposed to Trastuzumab and EC-Fc (EC-1 peptide conjugated to Fc part of human IgG). On the other hand, this caveolae dependent endocytic synergy was not observed in parental SKBR-3 cells. Therefore, caveolin-1 expression in breast cancer cells could be a predictive factor to estimate how cancer cells are likely to respond to Trastuzumab treatment.

    DOI: 10.7150/jca.6470

    Web of Science

    researchmap

  • Identification of Caveolin-1 as a Potential Causative Factor in the Generation of Trastuzumab Resistance in Breast Cancer Cells

    Sreeja C. Sekhar, Tomonari Kasai, Ayano Satoh, Tsukasa Shigehiro, Akifumi Mizutani, Hiroshi Murakami, Bishoy Y. A. El-Aarag, David S. Salomon, Anna Massaguer, Rafael de Llorens, Masaharu Seno

    JOURNAL OF CANCER   4 ( 5 )   391 - 401   2013

     More details

    Language:English   Publisher:IVYSPRING INT PUBL  

    The oncogenic tyrosine kinase receptor ErbB2 is a prognostic factor and target for breast cancer therapeutics. In contrast with the other ErbB receptors, ErbB2 is hardly internalized by ligand induced mechanisms, indicating a prevalent surface expression. Elevated levels of ErbB2 in tumor cells are associated with its defective endocytosis and down regulation. Here we show that caveolin-1 expression in breast cancer derived SKBR-3 cells (SKBR-3/Cav-1) facilitates ligand induced ErbB2 endocytosis using an artificial peptide ligand EC-eGFP. Similarly, stimulation with humanized anti ErbB2 antibody Trastuzumab (Herceptin) was found to be internalized and co-localized with caveolin-1 in SKBR-3/Cav-1 cells. Internalized EC-eGFP and Trastuzumab in SKBR-3/Cav-1 cells were then delivered via caveolae to the caveolin-1 containing early endosomes. Consequently, attenuated Fc receptor mediated ADCC functions were observed when exposed to Trastuzumab and EC-Fc (EC-1 peptide conjugated to Fc part of human IgG). On the other hand, this caveolae dependent endocytic synergy was not observed in parental SKBR-3 cells. Therefore, caveolin-1 expression in breast cancer cells could be a predictive factor to estimate how cancer cells are likely to respond to Trastuzumab treatment.

    DOI: 10.7150/jca.6470

    Web of Science

    researchmap

  • Eosinophil cationic protein enhances cardiomyocyte differentiation of P19CL6 embryonal carcinoma cells by stimulating the FGF receptor signaling pathway

    Guoliang Jin, Akifumi Mizutani, Takayuki Fukuda, Ling Chen, Keisuke Nakanishi, Ting Yan, Takayuki Kudoh, Satoshi Hirohata, Tomonari Kasai, Hiroshi Murakami, David S. Salomon, Masaharu Seno

    GROWTH FACTORS   30 ( 5 )   344 - 355   2012.10

     More details

    Language:English   Publisher:INFORMA HEALTHCARE  

    We investigated the functional role of eosinophil cationic protein (ECP) in regulating cardiomyogenesis using mouse P19CL6 embryonic carcinoma cells. ECP was confirmed to accelerate the cardiomyocyte differentiation of P19CL6 cells by enhancing the rate and area size of beating of cardiomyocyte and by facilitating the expression of cardiomyocyte-specific genes, such as GATA4 and alpha-MHC. Since cardiomyocyte differentiation in vivo is considered to follow mesoderm induction, the induction of Brachyury, a marker of mesoderm, was assessed. Brachyury expression was found to be enhanced after the addition of ECP. This enhancement was due to the stimulation of extracellular signal-regulated kinase (ERK) 1/2 phosphorylation by ECP. In this context, treatment with SU5402, an inhibitor of fibroblast growth factor (FGF) receptor 1, suppressed Brachyury expression, phosphorylation of ERK1/2, and cardiomyocyte differentiation induced by ECP. We concluded that ECP might induce mesoderm differentiation through FGF signaling pathway and enhance subsequent cardiomyocyte differentiation in concert with dimethyl sulfoxide in P19CL6 cells. ECP may be a novel factor for cardiomyocyte differentiation, which should be very useful to prepare adequate numbers of cardiomyocytes for therapeutic cell transplantation.

    DOI: 10.3109/08977194.2012.709852

    Web of Science

    researchmap

  • Eosinophil cationic protein enhances cardiomyocyte differentiation of P19CL6 embryonal carcinoma cells by stimulating the FGF receptor signaling pathway

    Guoliang Jin, Akifumi Mizutani, Takayuki Fukuda, Ling Chen, Keisuke Nakanishi, Ting Yan, Takayuki Kudoh, Satoshi Hirohata, Tomonari Kasai, Hiroshi Murakami, David S. Salomon, Masaharu Seno

    GROWTH FACTORS   30 ( 5 )   344 - 355   2012.10

     More details

    Language:English   Publisher:INFORMA HEALTHCARE  

    We investigated the functional role of eosinophil cationic protein (ECP) in regulating cardiomyogenesis using mouse P19CL6 embryonic carcinoma cells. ECP was confirmed to accelerate the cardiomyocyte differentiation of P19CL6 cells by enhancing the rate and area size of beating of cardiomyocyte and by facilitating the expression of cardiomyocyte-specific genes, such as GATA4 and alpha-MHC. Since cardiomyocyte differentiation in vivo is considered to follow mesoderm induction, the induction of Brachyury, a marker of mesoderm, was assessed. Brachyury expression was found to be enhanced after the addition of ECP. This enhancement was due to the stimulation of extracellular signal-regulated kinase (ERK) 1/2 phosphorylation by ECP. In this context, treatment with SU5402, an inhibitor of fibroblast growth factor (FGF) receptor 1, suppressed Brachyury expression, phosphorylation of ERK1/2, and cardiomyocyte differentiation induced by ECP. We concluded that ECP might induce mesoderm differentiation through FGF signaling pathway and enhance subsequent cardiomyocyte differentiation in concert with dimethyl sulfoxide in P19CL6 cells. ECP may be a novel factor for cardiomyocyte differentiation, which should be very useful to prepare adequate numbers of cardiomyocytes for therapeutic cell transplantation.

    DOI: 10.3109/08977194.2012.709852

    Web of Science

    researchmap

  • A Model of Cancer Stem Cells Derived from Mouse Induced Pluripotent Stem Cells

    Ling Chen, Tomonari Kasai, Yueguang Li, Yuh Sugii, Guoliang Jin, Masashi Okada, Arun Vaidyanath, Akifumi Mizutani, Ayano Satoh, Takayuki Kudoh, Mary J. C. Hendrix, David S. Salomon, Li Fu, Masaharu Seno

    PLOS ONE   7 ( 4 )   e33544   2012.4

     More details

    Language:English   Publisher:PUBLIC LIBRARY SCIENCE  

    Cancer stem cells (CSCs) are capable of continuous proliferation and self-renewal and are proposed to play significant roles in oncogenesis, tumor growth, metastasis and cancer recurrence. CSCs are considered derived from normal stem cells affected by the tumor microenvironment although the mechanism of development is not clear yet. In 2007, Yamanaka's group succeeded in generating Nanog mouse induced pluripotent stem (miPS) cells, in which green fluorescent protein (GFP) has been inserted into the 5'-untranslated region of the Nanog gene. Usually, iPS cells, just like embryonic stem cells, are considered to be induced into progenitor cells, which differentiate into various normal phenotypes depending on the normal niche. We hypothesized that CSCs could be derived from Nanog miPS cells in the conditioned culture medium of cancer cell lines, which is a mimic of carcinoma microenvironment. As a result, the Nanog miPS cells treated with the conditioned medium of mouse Lewis lung carcinoma acquired characteristics of CSCs, in that they formed spheroids expressing GFP in suspension culture, and had a high tumorigenicity in Balb/c nude mice exhibiting angiogenesis in vivo. In addition, these iPS-derived CSCs had a capacity of self-renewal and expressed the marker genes, Nanog, Rex1, Eras, Esg1 and Cripto, associated with stem cell properties and an undifferentiated state. Thus we concluded that a model of CSCs was originally developed from miPS cells and proposed the conditioned culture medium of cancer cell lines might perform as niche for producing CSCs. The model of CSCs and the procedure of their establishment will help study the genetic alterations and the secreted factors in the tumor microenvironment which convert miPS cells to CSCs. Furthermore, the identification of potentially bona fide markers of CSCs, which will help the development of novel anti-cancer therapies, might be possible though the CSC model.

    DOI: 10.1371/journal.pone.0033544

    Web of Science

    researchmap

  • Preparation and evaluation of glycosylated paclitaxel loaded in tastuzumab-immunoliposome

    Masaharu Murakami, Tomonari Kasai, Masashi Okada, Katsuhiko Mikuni, Naoyoshi Egashira, Masaharu Seno, Hiroki Hamada

    CANCER RESEARCH   72   2012.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2012-5700

    Web of Science

    researchmap

  • Development and characterization of cancer stem cell model from mouse iPS cells

    Ling Chen, Shuichi Matsuda, Tomonari Kasai, Yuh Sugii, Masashi Okada, Koichi Igarashi, Ayano Satoh, Takayuki Kudoh, Takayuki Kudoh, Li Fu, Masaharu Seno

    CANCER RESEARCH   72   2012.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2012-418

    Web of Science

    researchmap

  • The Conformational Polymorphism of the Green Fluorescent Protein

    Haidong Tan, Yueguang Li, Ling Chen, Takayuki Kudoh, Tomonari Kasai, Masaharu Seno

    MOLECULAR BIOLOGY   46 ( 1 )   142 - 148   2012.2

     More details

    Language:English   Publisher:MAIK NAUKA/INTERPERIODICA/SPRINGER  

    Green fluorescent protein (GFPuv) has been widely used as a reporter fused to individual targeting sequences. However, its state in liquid and its effect on other proteins are still unclear. The conformational polymorphisms of glutathione-S-transferase-green fluorescent protein (GST-GFPuv), GFPuv and GST were analyzed by native polyacrylamide gel, indicating that GST was in many different states while GFPuv and GST-GFPuv were only in four and two slightly different states. Four different circular dichroism spectra were obtained from the GFPuv polymorphisms. The single molecular behavior of GST-GFPuv and GFPuv was also characterized by MALDI-TOF MS. Thus, we demonstrated that: (1) there might be four different structural polymorphisms for the native GFPuv; (2) GFPuv could reduce its partner's polymorphism as a fusion protein. Although GFPuv had many merits as a reporter, its unreliability was found in the study. DOI: 10.1134/S0026893311060045

    DOI: 10.1134/S0026893311060045

    Web of Science

    researchmap

  • The Conformational Polymorphism of the Green Fluorescent Protein

    Haidong Tan, Yueguang Li, Ling Chen, Takayuki Kudoh, Tomonari Kasai, Masaharu Seno

    MOLECULAR BIOLOGY   46 ( 1 )   142 - 148   2012.2

     More details

    Language:English   Publisher:MAIK NAUKA/INTERPERIODICA/SPRINGER  

    Green fluorescent protein (GFPuv) has been widely used as a reporter fused to individual targeting sequences. However, its state in liquid and its effect on other proteins are still unclear. The conformational polymorphisms of glutathione-S-transferase-green fluorescent protein (GST-GFPuv), GFPuv and GST were analyzed by native polyacrylamide gel, indicating that GST was in many different states while GFPuv and GST-GFPuv were only in four and two slightly different states. Four different circular dichroism spectra were obtained from the GFPuv polymorphisms. The single molecular behavior of GST-GFPuv and GFPuv was also characterized by MALDI-TOF MS. Thus, we demonstrated that: (1) there might be four different structural polymorphisms for the native GFPuv; (2) GFPuv could reduce its partner's polymorphism as a fusion protein. Although GFPuv had many merits as a reporter, its unreliability was found in the study. DOI: 10.1134/S0026893311060045

    DOI: 10.1134/S0026893311060045

    Web of Science

    researchmap

  • Chlorotoxin Fused to IgG-Fc Inhibits Glioblastoma Cell Motility via Receptor-Mediated Endocytosis.

    2012   975763   2012

     More details

  • A model of cancer stem cells derived from mouse induced pluripotent stem cells.

    PLoS One   7 ( 4 )   e33544   2012

     More details

    Language:English  

    researchmap

  • Chlorotoxin Fused to IgG-Fc Inhibits Glioblastoma Cell Motility via Receptor-Mediated Endocytosis.

    2012   975763   2012

     More details

  • Enhanced internalization of ErbB2 in SK-BR-3 cells with multivalent forms of an artificial ligand

    Arun Vaidyanath, Toshihiro Hashizume, Tadahiro Nagaoka, Nao Takeyasu, Hitomi Satoh, Ling Chen, Jiyou Wang, Tomonari Kasai, Takayuki Kudoh, Ayano Satoh, Li Fu, Masaharu Seno

    JOURNAL OF CELLULAR AND MOLECULAR MEDICINE   15 ( 11 )   2525 - 2538   2011.11

     More details

    Language:English   Publisher:WILEY-BLACKWELL  

    Targeting and down-regulation of ErbB2, a member of EGF receptor family, is regarded as one of the key aspect for cancer treatment because it is often overexpressed in breast and ovarian cancer cells. Although natural ligands for ErbB2 have not been found, unlike other ErbB receptors, EC-1, a 20-amino acid circular peptide, has been shown to bind to ErbB2 as an artificial ligand. Previously we showed EC-1 peptide did not induce the internalization of ErbB2 in SK-BR-3 cells. In this report, we designed divalent and multivalent forms of EC-1 peptide with the Fc portion of the human IgG and bionanocapsule modified with ZZ-tag on its surface to improve the interaction with ErbB2. These forms showed higher affinity to ErbB2 than that of EC-1 monomer. Furthermore, prominent endosomal accumulation of ErbB2 occurred in SK-BR-3 cells when stimulated with EC-Fc ligand multivalently displayed on the surface of the bionanocapsule, whereas SK-BR-3 cells as themselves displayed stringent mechanism against ErbB2 internalization without stimulation. The multivalent form of EC-1 peptide appeared to internalize ErbB2 more efficiently than divalent form did. This internalization was unaffected by the inhibition of clathrin association, but inhibited when the cholesterol was depleted which explained either caveolar or GPI-AP-early endocytic compartment (GEEC) pathway. Because of the lack of caveolin-1 expression, caveolar machinery may be lost in SK-BR-3 cell line. Therefore, it is suggested that the multivalent form of EC-1 induces the internalization of ErbB2 through the GEEC pathway.

    DOI: 10.1111/j.1582-4934.2011.01277.x

    Web of Science

    researchmap

  • Enhanced internalization of ErbB2 in SK-BR-3 cells with multivalent forms of an artificial ligand

    Arun Vaidyanath, Toshihiro Hashizume, Tadahiro Nagaoka, Nao Takeyasu, Hitomi Satoh, Ling Chen, Jiyou Wang, Tomonari Kasai, Takayuki Kudoh, Ayano Satoh, Li Fu, Masaharu Seno

    JOURNAL OF CELLULAR AND MOLECULAR MEDICINE   15 ( 11 )   2525 - 2538   2011.11

     More details

    Language:English   Publisher:WILEY-BLACKWELL  

    Targeting and down-regulation of ErbB2, a member of EGF receptor family, is regarded as one of the key aspect for cancer treatment because it is often overexpressed in breast and ovarian cancer cells. Although natural ligands for ErbB2 have not been found, unlike other ErbB receptors, EC-1, a 20-amino acid circular peptide, has been shown to bind to ErbB2 as an artificial ligand. Previously we showed EC-1 peptide did not induce the internalization of ErbB2 in SK-BR-3 cells. In this report, we designed divalent and multivalent forms of EC-1 peptide with the Fc portion of the human IgG and bionanocapsule modified with ZZ-tag on its surface to improve the interaction with ErbB2. These forms showed higher affinity to ErbB2 than that of EC-1 monomer. Furthermore, prominent endosomal accumulation of ErbB2 occurred in SK-BR-3 cells when stimulated with EC-Fc ligand multivalently displayed on the surface of the bionanocapsule, whereas SK-BR-3 cells as themselves displayed stringent mechanism against ErbB2 internalization without stimulation. The multivalent form of EC-1 peptide appeared to internalize ErbB2 more efficiently than divalent form did. This internalization was unaffected by the inhibition of clathrin association, but inhibited when the cholesterol was depleted which explained either caveolar or GPI-AP-early endocytic compartment (GEEC) pathway. Because of the lack of caveolin-1 expression, caveolar machinery may be lost in SK-BR-3 cell line. Therefore, it is suggested that the multivalent form of EC-1 induces the internalization of ErbB2 through the GEEC pathway.

    DOI: 10.1111/j.1582-4934.2011.01277.x

    Web of Science

    researchmap

  • Construction of a high-efficiency multi-site-directed mutagenesis

    Haidong Tan, Yueguang Li, Ling Chen, Tomonari Kasai, Masaharu Seno

    AFRICAN JOURNAL OF BIOTECHNOLOGY   10 ( 3 )   449 - 452   2011.1

     More details

    Language:English   Publisher:ACADEMIC JOURNALS  

    Although site-directed mutagenesis has been used in many fields, it still has low rate of success and high cost because of low-yield target products. A modified method for multi-site-directed mutagenesis was developed with shifted primer design and cold-start polymerase chain reaction (PCR). The developed method was successfully applied to hexapeptide gene synthesis and recombinant enterokinase gene modification in the plasmids pET41a and pET24b-EK. The efficiency was pronounced at a 1:10 molar ratio of 7-base mutant products to 705-bp fragment products as control. Even in a 10-base substitution mutagenic PCR, a 1:50 molar ratio of mutant products to 705-bp fragment products was reached. Meanwhile, the quality of mutants was proved through the transformation efficiency and sequencing. This method was beneficial to prepare high-quality multibase mutagenesis and also implied that large-scale multibase mutagenesis was feasible, efficient, economical, and productive.

    Web of Science

    researchmap

  • Extracellular Matrix Modulates Insulin Production During Differentiation of AR42J Cells: Functional Role of Pax6 Transcription Factor

    Kohei Hamamoto, Satoko Yamada, Akemi Hara, Tsutomu Kodera, Masaharu Seno, Itaru Kojima

    JOURNAL OF CELLULAR BIOCHEMISTRY   112 ( 1 )   318 - 329   2011.1

     More details

    Language:English   Publisher:WILEY-BLACKWELL PUBLISHING, INC  

    Extracellular matrix (ECM) modulates differentiation of pancreatic beta-cells during development. However, the mechanism by which ECM proteins modulate differentiation is not totally clear. We investigated the effect of ECM proteins on differentiation beta-cells in vitro. We investigated the effect of basement membrane ECM on differentiation of AR42J cells and rat ductal cells. First, we examined the effect of reconstituted basement membrane, Matrigel on differentiation of AR42J cells induced by activin and betacellulin. Matrigel augmented insulin production and increased the expression of GLUT2, SUR1, and glucokinase. Among various transcription factors investigated, Matrigel markedly upregulated the expression of Pax6. When Pax6 was overexpressed in cells treated with activin and betacellulin, the expression of insulin was upregulated. Conversely, knockdown of Pax6 significantly reduced the insulin expression in cells cultured on Matrigel. The effects of Matrigel on insulin-production and induction of Pax6 were reproduced partially by laminin-1, a major component of Matrigel, and inhibited by anti-integrin-beta 1 antibody. Matrigel also enhanced the activation of p38 mitogen-activated kinase induced by activin and betacellulin, which was inhibited by anti-beta 1 antibody. Finally, the effect of Matrigel on differentiation was reproduced in rat cultured ductal cells, and Matrigel also increased the expression of Pax6. These results indicate that basement membrane ECM augments differentiation of pancreatic progenitor cells to insulin-secreting cells by upregulating the expression of Pax6. J. Cell. Biochem. 112: 318329, 2011. (C) 2010 Wiley-Liss, Inc.

    DOI: 10.1002/jcb.22930

    Web of Science

    researchmap

  • Clustering Genes, Tissues, Cells and Bioactive Chemicals by Sphere SOM

    Self Organizing Maps-applications and novel algorithm design   15 ( 11 )   371 - 386   2011

     More details

  • Clustering Genes, Tissues, Cells and Bioactive Chemicals by Sphere SOM

    15 ( 11 )   371 - 386   2011

     More details

  • Analysis of endothelium mimicry by the CSC-like cells derived from iPS cells.

    S-I. Matsuda, A. Mizutani, T. Kasai, A. Satoh, T. Kudoh, L. Chen, M. Seno

    MOLECULAR BIOLOGY OF THE CELL   22   2011

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Generation of Cancer Stem Cell Model from Mouse iPS Cells.

    A. Mizutani, S-I. Matsuda, T. Kasai, T. Kudoh, L. Chen, M. Seno

    MOLECULAR BIOLOGY OF THE CELL   22   2011

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Optimization of Bacterial Plasmid Transformation Using Nanomaterials Based on the Yoshida Effect

    Haidong Tan, Li Fu, Masaharu Seno

    INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES   11 ( 12 )   4962 - 4973   2010.12

     More details

    Language:English   Publisher:MDPI AG  

    With the help of sepiolite, a unique method for transforming DNA into bacteria, based on the Yoshida effect, has been developed recently. However, we confronted many problems when this newest method was tried. Only a few transformants could be obtained even when 100 ng of plasmid pET15b was used, and a successful result seemed difficult to repeat. To address this problem, we optimized the operating method and could achieve about 15,000 transformants using the same amount of plasmid, which could match the efficiency gained using the calcium chloride transformation method. Meanwhile, the results could also be reproduced well. In the same way, carbon nanotubes were used to attain more than 15,000 transformants in the same situation. Therefore, the transformation method could be extended to other nanomaterials. Meanwhile, compared with the mechanism previously reported, we verified quite a different principle for the mechanism responsible for such a transformation. In sum, this unique transformation can be developed to become the third widely-used transformation method in laboratories in addition to the chemical method and electroporation.

    DOI: 10.3390/ijms11124962

    Web of Science

    researchmap

  • First Report of Cucumber mosaic virus in Sweet Cherry in the People&apos;s Republic of China

    H. D. Tan, S. Y. Li, X. F. Du, M. Seno

    PLANT DISEASE   94 ( 11 )   1378 - 1378   2010.11

     More details

    Language:English   Publishing type:Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)   Publisher:AMER PHYTOPATHOLOGICAL SOC  

    DOI: 10.1094/PDIS-07-10-0549

    Web of Science

    researchmap

  • Novel and simple loading procedure of cisplatin into liposomes and targeting tumor endothelial cells

    M. Hirai, H. Minematsu, Y. Hiramatsu, H. Kitagawa, T. Otani, S. Iwashita, T. Kudoh, L. Chen, Y. Li, M. Okada, D. S. Salomon, K. Igarashi, M. Chikuma, M. Seno

    INTERNATIONAL JOURNAL OF PHARMACEUTICS   391 ( 1-2 )   274 - 283   2010.5

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    Although intravenous administration of high levels of cisplatin (CDDP) are limited due to its severe side effects, efficient delivery of CDDP directly to the tumor should improve the therapeutic response while potentially by-passing significant side effects.
    High loading of CDDP into liposomes is one technique that could be used as a potential drug delivery system. Since cis-diamminedinitratoplatinum (CDDP3) is highly soluble in water and converts to CDDP in the presence of chloride ions, we encapsulated CDDP3 into liposomes in the absence of chloride ions and supplemented chloride ions to prepare CDDP-encapsulated liposomes (CDDP-Lip) resulting in a significantly improved loading efficiency of COOP. We further conjugated the CDDP-Lip with Sialyl Lewis(X) (CDDP-SLX-Lip) because we previously demonstrated Sialyl Lewis(X) enhanced efficient accumulation of liposomes into tumors in vivo. CDDP-SLX-Lip treated mice showed a survival rate of 75% at 14 days even if a lethal level of CDDP was injected into mice. Loss of body weight was negligible and no histological abnormality was found in a variety of normal tissues. Accumulation of CDDP-SLX-Lip was about 6 times more than that of CDDP-Lip or CDDP. As the result, there was better antitumor activity of CDDP-SLX-Lip than that of CDDP-Lip with significantly less toxic effects in normal tissues. (C) 2010 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.ijpharm.2010.02.030

    Web of Science

    researchmap

  • E-selectin targeting to visualize tumors in vivo

    Masahiko Hirai, Yoshie Hiramatsu, Shinki Iwashita, Takayuki Otani, Ling Chen, Yue-guang Li, Masashi Okada, Kazunori Oie, Koich Igarashi, Hideaki Wakita, Masaharu Seno

    CONTRAST MEDIA & MOLECULAR IMAGING   5 ( 2 )   70 - 77   2010.3

     More details

    Language:English   Publisher:JOHN WILEY & SONS LTD  

    Generally angiogenic factors induce the expression of E-selectin in vascular endothelial cells in the tumors. In this study, we employed an anti-E-selectin monoclonal antibody to target tumors in vivo and evaluated an optical imaging reagent to visualize tumor regions. The anti-E-selectin antibody was conjugated on the surface of liposomes, which encapsulated the near-infrared fluorescent substances Cy3 or Cy5.5. The liposomes efficiently recognized human umbilical vein endothelial cells only when E-selectin was induced by angiogenic factors such as TNF-alpha in vitro. Cy5.5 encapsulated into liposomes that were conjugated with an anti-E-selectin antibody successfully visualized Ehrlich ascites tumor cells when transplanted into mice. Thus, E-selectin targeting with liposomes containing a near-infrared fluorescent dye was found effective in visualizing tumors in vivo. This strategy should be extremely useful as a method to identify sentinel lymphatic nodes and angiogenic tumors as well as use for drug delivery to tumor cells. Copyright (C) 2010 John Wiley & Sons, Ltd.

    DOI: 10.1002/cmmi.367

    Web of Science

    researchmap

  • Production of biologically active IgG hinge-tag soluble epidermal growth factor receptors (ErbB)

    Takayuki Otani, Toshihiro Hashizume, Tadahiro Nagaoka, Tomoko Fukuda, Careen K. Tang, David S. Salomon, Masaharu Seno

    BIOTECHNOLOGY LETTERS   32 ( 3 )   361 - 366   2010.3

     More details

    Language:English   Publisher:SPRINGER  

    The extracellular domains (ECD) of epidermal growth factor receptors, ErbB1, 2, 3 and 4, were designed as soluble dimeric forms. Each ECD was fused to a short hinge region derived from IgG, such that the stable dimer could be formed with disulfide bridges. This hinge-tagged design minimized the molecular weight to approximately 50% of the conventional Fc-fusion design without an Fc domain of IgG. The refolded dimers could be easily analyzed and characterized by SDS-PAGE. Hinge-tagged soluble ErbBs demonstrated significant affinity for betacellulin and heregulin. The IgG hinge-tag should be a simple method to design soluble dimers that would be useful for high throughput screening of ligands, antagonists or derivatives.

    DOI: 10.1007/s10529-009-0160-9

    Web of Science

    researchmap

  • Administration of Conophylline and Betacellulin-delta 4 Increases the beta-cell Mass in Neonatal Streptozotocin-treated Rats

    Tsutomu Kodera, Satoko Yamada, Yoritsuna Yamamoto, Akemi Hara, Yuji Tanaka, Masaharu Seno, Kazuo Umezawa, Izumi Takei, Itaru Kojima

    ENDOCRINE JOURNAL   56 ( 6 )   799 - 806   2009.9

     More details

    Language:English   Publisher:JAPAN ENDOCRINE SOC  

    The present study was conducted to examine the effect of administration of conophylline (CnP) and betacellulin delta 4 (BTC delta 4) on the beta-cell mass in neonatal streptozotocin-treated rats (neonatal STZ rats). STZ (100 mu g/g) was injected into neonatal rats, and then CnP (2 mu g/g) and/or BTC delta 4 (200 pmol/g) were administered to neonatal STZ rats for 1 week. The plasma glucose concentration was monitored, and an intraperitoneal glucose tolerance test (ipGTT) was performed on day 8 and at 8 weeks after the STZ injection. In neonatal STZ rats treated with control solution (S group), the plasma glucose concentration increased for several days after the STZ injection, returned to nearly normal levels, and then increased gradually after six weeks of age. Eight weeks after the STZ-injection, the plasma glucose concentration was increased significantly compared to that of normal rats. The glucose response to ipGTT was significantly reduced in neonatal STZ rats treated with CnP (CnP group), BTC delta 4 (delta 4 group) and CnP+BTC delta 4 (CnP+delta 4 group). The beta-cell mass and the insulin content of the pancreas were significantly increased in the CnP group and delta 4 group. The effect of CnP+delta 4 was greater than ;that of CnP alone or BTC delta 4 alone. CnP+BTC delta 4 significantly increased the number of PDX-1-positive ductal cells and the number of insulin/BrdU double-positive ductal cells. These results indicate the efficacy of CnP and BTC delta 4 in increasing the beta-cells mass of neonatal STZ-treated rats.

    DOI: 10.1507/endocrj.K09E-158

    Web of Science

    researchmap

  • Delivery of sodium borocaptate to glioma cells using immunoliposome conjugated with anti-EGFR antibodies by ZZ-His

    Bin Feng, Kazuhito Tomizawa, Hiroyuki Michiue, Shin-ichi Miyatake, Xiao-Jian Han, Atsushi Fujimura, Masaharu Seno, Mitsunori Kirihata, Hideki Matsui

    BIOMATERIALS   30 ( 9 )   1746 - 1755   2009.3

     More details

    Language:English   Publisher:ELSEVIER SCI LTD  

    Nanoparticles are effective of delivering cargo into cells. Here, sodium borocaptate (BSH) was encapsulated in liposomes composed of nickel lipid, and anti-epidermal growth factor receptor (EGFR) antibodies were conjugated to the liposomes using the antibody affinity motif of protein A (ZZ) as an adaptor (immunoliposomes). The immunoliposomes were used to deliver BSH into EGFR-overexpressing glioma cells. Immunohistochemical analysis using an anti-BSH monoclonal antibody revealed that BSH was delivered effectively into the cells but not into EGFR-deficient glioma or primary astrocytes. In an animal model of brain tumors, both the liposomes and the BSH were only observed in the tumor. Moreover, the efficiency of (10)B&apos;s delivery into glioma cells was confirmed by inductively coupled plasma-atomic emission spectrometry (ICP-AES) both in vitro and in vivo. The results suggest that this system utilizing immunoliposomes provides an effective means of delivering (10)B into glioma cells in boron neutron capture therapy (BNCT). (C) 2008 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.biomaterials.2008.12.010

    Web of Science

    researchmap

  • Human eosinophil cationic protein enhances stress fiber formation in Balb/c 3T3 fibroblasts and differentiation of rat neonatal cardiomyocytes

    Takayuki Fukuda, Miki Iwata, Midori Kitazoe, Takashi Maeda, David Salomon, Satoshi Hirohata, Katsuyuki Tanizawa, Shun&apos;ichi Kuroda, Masaharu Seno

    GROWTH FACTORS   27 ( 4 )   228 - 236   2009

     More details

    Language:English   Publisher:TAYLOR & FRANCIS LTD  

    We found that eosinophil cationic protein (ECP) stimulated the growth of mouse Balb/c 3T3 fibroblasts. ECP-treated 3T3 cells were more flattened and exhibited enhanced stress fiber formation. The enhancement of cytoskeleton after addition of recombinant ECP appeared stable and was able to inhibit disassembly of actin filaments that was induced by fibroblast growth factor-2. The ROCK inhibitor, Y-27632, abrogated this enhancement on stress fiber formation that was induced by ECP indicating the involvement of Rho/ROCK signaling pathway. The effect of ECP was assessed on the differentiation of primary cardiomyocytes derived from rat neonatal heart since the development of actin filaments is significantly related with organization of stress fibers. As the result, both beating rate and the expression of cardiac muscle specific markers such as atrial natriuretic factor were enhanced in the presence of ECP. Thus ECP may also function as a cardiomyocyte differentiation factor.

    DOI: 10.1080/08977190902987149

    Web of Science

    researchmap

  • Spherical self-organizing map as a helpful tool to identify category-specific cell surface markers

    Tuoya, Yuh Sugii, Hitomi Satoh, Dongwei Yu, Yasaburo Matsuura, Heizo Tokutaka, Masaharu Seno

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   376 ( 2 )   414 - 418   2008.11

     More details

    Language:English   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We analyzed gene expression profiles of five tumor cell lines (NB2a, NB41A3, C1300N18, BC3H1, and Neuro2a) derived from a category of nervous system using our originally developed cell surface marker DNA microarray in order to search for tumor-specific cell surface markers common to these cells. To visualize the expression patterns and to extract candidate genes of interest based on the expression profiles of several cell lines, we employed the clustering procedure of spherical self-organizing-map. As the result, three candidates of tumor-specific cell surface markers were picked up when the expression profiles were compared with that from normal brain tissue. RT-qPCR showed the expression of these genes was higher in tumor cells than in normal brain. Here we demonstrated the spherical self-organizing-map analysis should be useful to identify the candidates of cell surface markers common and specific to the group of cells or tissues of interest. (C) 2008 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2008.09.010

    Web of Science

    researchmap

  • Intracellular delivery of glutathione S-transferase-fused proteins into mammalian cells by polyethylenimine-glutathione conjugates

    Hitoshi Murata, Junichiro Futami, Midori Kitazoe, Takayuki Yonehara, Hidetaka Nakanishi, Megumi Kosaka, Hiroko Tada, Masakiyo Sakaguchi, Yasuyuki Yagi, Masaharu Seno, Nam-ho Huh, Hidenori Yamada

    JOURNAL OF BIOCHEMISTRY   144 ( 4 )   447 - 455   2008.10

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    The glutathione S-transferase (GST)-fused protein expression system has been extensively used to generate a large quantity of proteins and has served for functional analysis in vitro. In this study, we developed a novel approach for the efficient intracellular delivery of GST-fused proteins into living cells to expand their usefulness up to in vivo use. Since protein cationization techniques are powerful strategies for efficient intracellular uptake by adsorptive-mediated endocytosis, GST-fused proteins were cationized by forming a complex with a polycationic polyethylenimine (PEI)-glutathione conjugate. On screening of protein transduction, optimized PEI-glutathione conjugate for protein transduction was characterized by a partly oligomerized mixture of PEI with average molecular masses of 600 (PEI600) modified with multiple glutathiones, which could have sufficient avidity for GST. Furthermore, enhanced endosomal escape of transduced GST-fused proteins was observed when they were delivered with a glutathione-conjugated PEI600 derivative possessing a hydroxybutenyl moiety. These results were confirmed by both intracellular confocal imaging of GST-fused green fluorescent protein and activation of an endogenous growth signal transduction pathway by a GST-fused constitutively active mutant of a kinase protein. These PEI-glutathione conjugates seem to be convenient molecular tools for protein transduction of widely used GST-fused proteins.

    DOI: 10.1093/jb/mvn087

    Web of Science

    researchmap

  • Smad2 functions as a co-activator of canonical Wnt/beta-catenin signaling pathway independent of Smad4 through histone acetyltransferase activity of p300

    Morihisa Hirota, Kazuhide Watanabe, Shin Hamada, Youping Sun, Luigi Strizzi, Mario Mancino, Tadahiro Nagaoka, Monica Gonzales, Masaharu Seno, Caterina Bianco, David S. Salomon

    CELLULAR SIGNALLING   20 ( 9 )   1632 - 1641   2008.9

     More details

    Language:English   Publisher:ELSEVIER SCIENCE INC  

    Both canonical Wnt/beta-catenin and TGF beta/Smad signaling pathways coordinately regulate pattern formation during embryogenesis as well as tumor progression. Evidence of cross-talk between these two pathways has been reported. Here we demonstrated that the Activin-like kinase 4 (Alk4)/Smad2 pathway facilitates the transcriptional activity of the oncogenic Wnt/beta-catenin/Tcf4 pathway through a novel Smad4-independent mechanism. Upon activation, Smad2 physically interacted with Tcf4, beta-catenin and the co-activator p300 to enhance transcriptional activity of beta-catenin/Tcf4 through the histone acetyltransferase activity of p300. Transactivation by Smad2 was independent of a Smad-binding element (SBE) and Smad4. Indeed, the enhancement of beta-catenin/Tcf4 transcriptional activity by activated Smad2 was negatively regulated by the presence of Smad4. Moreover, a tumor-derived missense mutant of Smad2, lacking the ability to bind to Smad4 was still able to enhance the Tcf4 transcriptional reporter in the presence of p-catenin and Tcf4. Our findings suggest that Smad2 may function as an activator of canonical Wnt/beta-catenin/Tcf4 signaling through a SBE/Smad4-independent pathway. Published by Elsevier Inc.

    DOI: 10.1016/j.cellsig.2008.05.003

    Web of Science

    researchmap

  • Characteristics of hollow microtubes consisting of amorphous iron oxide nanoparticles produced by iron oxidizing bacteria, Leptothrix ochracea (vol 310, pg 2405, 2007)

    H. Hashimoto, S. Yokoyama, H. Asaoka, Y. Kusano, Y. Ikeda, M. Seno, J. Takada, T. Fujii, M. Nakanishi, R. Murakami

    JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS   320 ( 18 )   2310 - 2310   2008.9

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    DOI: 10.1016/j.jmmm.2008.01.046

    Web of Science

    researchmap

  • Reversal of streptozotocin-induced hyperglycemia by continuous supply of betacellulin in mice

    Yoritsuna Yamamoto, Satoko Yamada, Tsutomu Kodera, Akemi Hara, Kazuo Motoyoshi, Yuji Tanaka, Tadahiro Nagaoka, Masaharu Seno, Itaru Kojima

    GROWTH FACTORS   26 ( 4 )   173 - 179   2008.8

     More details

    Language:English   Publisher:TAYLOR & FRANCIS LTD  

    Previous studies have shown the efficacy of betacellulin (BTC) to promote P-cell regeneration. Because of its short half-life, however, the effect of BTC may have been underestimated. This study was conducted to assess the effect of continuous administration of BTC on P-cell regeneration. Adenovirus vectors encoding proBTC (Ad-proBTC) and mature BTC (Ad-mBTC) were prepared, and the efficacy of secretion of BTC was compared in AML12 hepatocytes. When AML12 cells were infected with Ad-proBTC or Ad-mBTC, cells infected with Ad-mBTC secreted considerably larger amount of BTC. We then infused Ad-mBTC into the mouse tail vein. Expression of BTC was detected in the liver for at least 21 days, and serum BTC was maintained at approximately 1 ng/ml for 7 days. When Ad-mBTC was infused immediately after administration of STZ (170 mg/kg), elevation of the plasma glucose induced by STZ was markedly inhibited, and the plasma glucose concentration remained at less than 200 mg/dl for 21 days. The insulin content and the P-cell mass were significantly increased in Ad-mBTC-infused mice. These results indicate that continuous administration of BTC is quite effective in promoting regeneration of P-cells.

    DOI: 10.1080/08977190802136854

    Web of Science

    researchmap

  • Cell type dependent endocytic internalization of ErbB2 with an artificial peptide ligand that binds to ErbB2

    Toshihiro Hashizume, Takayuki Fukuda, Tadahiro Nagaoka, Hiroko Tada, Hidenori Yamada, Kazuhide Watanabe, David S. Salomon, Masaharu Seno

    CELL BIOLOGY INTERNATIONAL   32 ( 7 )   814 - 826   2008.7

     More details

    Language:English   Publisher:ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD  

    ErbB2, which is a member of the epidermal growth factor (erbB) receptor family, is frequently overexpressed in breast and ovarian cancers. Antibody and small molecule anti-tyrosine kinase inhibitors have been developed for targeted therapies for cancers overexpressing erbB2. Internalization and downregulation of erbB2, which is induced by a ligand, may be important for efficacious therapeutic effects. However, ligand-dependent erbB2 internalization has not been well characterized. Here we investigated the internalization of erbB2 in SKBr3 and SKOv3 cells, both overexpressing erbB2, using an EC-1 peptide fused to eGFP (EC-eGFP), which specifically binds to erbB2. ErbB2 was internalized in SKOv3 cells when the cells were treated with EC-eGFP. The accumulation of endosomal erbB2 was EC-eGFP dependent, which colocalized with transferrin implying endocytosis via clathrin-coated pits. In contrast, internalization of erbB2 was not observed in SKBr3 cells. As a result, two different mechanisms, which are cell type dependent for the internalization of erbB2, are proposed. (C) 2008 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.cellbi.2008.03.012

    Web of Science

    researchmap

  • A betacellulin mutant promotes differentiation of pancreatic acinar AR42J cells into insulin-producing cells with low affinity of binding to ErbB1

    Tadahiro Nagaoka, Takayuki Fukuda, Toshihiro Hlashizume, Tomoko Nishiyama, Hiroko Tada, Hidenori Yamada, David S. Salomon, Satoko Yamada, Itaru Kojima, Masaharu Seno

    JOURNAL OF MOLECULAR BIOLOGY   380 ( 1 )   83 - 94   2008.6

     More details

    Language:English   Publisher:ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD  

    Betacellulin (BTC) is one of the members of the epidermal growth factor (EGF) ligand family of ErbB receptor tyrosine kinases. It is a differentiation factor as well as a potent mitogen. BTC promotes the differentiation of pancreatic acinar-derived AR42J cells into insulin-producing cells. It independently and preferentially binds to two type I tyrosine kinase receptors, the EGF receptor (ErbB1) and ErbB4. However, the physiochemical characteristics of BTC that are responsible for its preferential binding to these two receptors have not been fully defined. In this study, to investigate the essential amino acid residues of BTC for binding to the two receptors, we introduced point mutations into the EGF domain of BTC employing error-prone PCR. The receptor binding abilities of 190 mutants expressed in Escherichia coli were assessed by enzyme immunoassay. Replacement of the glutamic acid residue at position 88 with a lysine residue in BTC was found to produce a significant loss of affinity for binding to ErbB1, while the affinity of binding to ErbB4 was unchanged. In addition, the mutant of BTC-E/88/K showed less growth-promoting activity on BALB/c 3T3 cells compared with that of the wild-type BTC protein. Interestingly, the BTC mutant protein promoted differentiation of pancreatic acinar AR42J cells at a high frequency into insulin-producing cells compared with AR42J cells that were treated with wild-type BTC protein. These results indicate the possibility of designing BTC mutants, which have an activity of inducing differentiation only, without facilitating growth promotion. (C) 2008 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.jmb.2008.03.054

    Web of Science

    researchmap

  • A simple method to induce differentiation of murine bone marrow mesenchymal cells to insulin-producing cells using conophylline and betacellulin-delta4

    Etsuko Hisanaga, Kee-Yong Park, Satoko Yamada, Hiromi Hashimoto, Toshlyukj Takeuchi, Masatomo Mori, Masaharu Seno, Kazuo Umezawa, Izumi Takei, Itaru Kojima

    ENDOCRINE JOURNAL   55 ( 3 )   535 - 543   2008.6

     More details

    Language:English   Publisher:JAPAN ENDOCRINE SOC  

    The present study was conducted to establish a method to induce differentiation of bone marrow (MB)-derived mesenchymal cells into insulin-producing cells. When mouse BM-derived mesenchymal cells were cultured for 60 days in medium containing 10% fetal calf serum and 25 mM glucose, they expressed insulin. Addition of activin A and betacellulin (BTC) accelerated differentiation, and immunoreactive insulin was detected 14 days after the treatment. Insulin-containing secretory granules were observed in differentiated cells by electron microscopy. Treatment of BM-derived mesenchymal cells with conophylline (CnP) and BTC-delta4 further accelerated differentiation, and mRNA for insulin was detected 5 to 7 days after the treatment. Mesencymal cells treated with CnP and BTC-delta4 responded to a high concentration of glucose and secreted mature insulin. When these cells were transplanted into streptozotocin-treated mice, they markedly reduced the plasma glucose concentration, and the effect continued for at least 4 weeks. These results indicate an efficacy of the combination of CnP and BTC-delta4 in inducing differentiation of BM-derived mesenchymal cells into insulin-producing cells.

    DOI: 10.1507/endocrj.K07E-173

    Web of Science

    researchmap

  • Involvement of MAPK signaling molecules and Runx2 in the NELL1-induced osteoblastic differentiation

    Nobuyuki Bokui, Takayuki Otani, Koichi Igarashi, Junichiro Kaku, Mitsuo Oda, Tadahiro Nagaoka, Masaharu Seno, Kenji Taternatsu, Toshihide Okajima, Takashi Matsuzaki, Kang Ting, Katsuyuki Tanizawa, Shunichi Kuroda

    FEBS LETTERS   582 ( 2 )   365 - 371   2008.1

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    NELL1 is an extracellular protein inducing osteogenic differentiation and bone formation of osteoblastic cells. To elucidate the intracellular signaling cascade evoked by NELL1, we have shown that NELL1 protein transiently activates the MAPK signaling cascade, induces the phosphorylation of Runx2, and promotes the rapid intracellular accumulation of Tyr-phosphorylated proteins. Unlike BMP2, NELL1 protein does not activate the Smad signaling cascade. These findings suggest that upon binding to a specific receptor NELL1 transduces an osteogenic signal through activation of certain Tyr-kinases associated with the Ras-MAPK cascade, and finally leads to the osteogenic differentiation. (C) 2007 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.febslet.2007.12.006

    Web of Science

    researchmap

  • Transient cell proliferation with polyethylenimine-cationized N-terminal domain of simian virus 40 large T-antigen

    Hitoshi Murata, Junichiro Futami, Midori Kitazoe, Megumi Kosaka, Hiroko Tada, Masaharu Seno, Hidenori Yamada

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   105 ( 1 )   34 - 38   2008.1

     More details

    Language:English   Publisher:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    Polyethylenimine (PEI) cationization is a powerful strategy for protein transduction into cells. In this study, we attempted the artificial regulation of cell proliferation by protein transduction of the N-terminal domain (1-132 amino acids) of the simian virus 40 large T-antigen (SVLT-N), which inactivates retinoblastoma family proteins but not p53. To deliver SVLT-N into cells, we employed an indirect cationization method by forming a complex of biotynylated SVLT-N through disulfide bonds (biotin-SS-SVLT-N) and PEI-cationized avidin (PEI600-avidin). Using this complex, SVLT-N was transduced into the nucleus of confluent and quiescent Balb/c 3T3 cells and was found to be complexed with a cellular target protein, pRb. Furthermore, SVLT-N transduction induced cell proliferation in spite of confluent conditions. Because SVLT-N thus transduced into cells gradually degraded and was not detectable after a 4-d incubation, transiently transformed cells were obtained by this method. These results suggest that oncogene protein transduction technology has great potential for in vitro regulation of cell proliferation.

    DOI: 10.1263/jbb.105.34

    Web of Science

    researchmap

  • Morphological and Microstructural Study of Iron Oxide Microtubes formed by Iron Oxidizing Bacteria, Leptothrix ochracea

    H. Hashimoto, H. Asaoka, J. Takada, T. Fujii, M. Nakanishi, S. Yokoyama, R. Murakami, Y. Kusano, Y. Ikeda, M. Seno

    Global Roadmap of Ceramics - ICC2 Proceedings   6-P-011-1-6-P-011-4   2008

     More details

  • Growth factor induction of cripto-1 shedding by glycosylphosphatidylinositol-phospholipase D and enhancement of endothelial cell migration

    Kazuhide Watanabe, Caterina Bianco, Luigi Strizzi, Shin Hamada, Mario Mancino, Veronique Bailly, Wenjun Mo, Dingyi Wen, Konrad Miatkowski, Monica Gonzales, Michele Sanicola, Masaharu Seno, David S. Salomon

    JOURNAL OF BIOLOGICAL CHEMISTRY   282 ( 43 )   31643 - 31655   2007.10

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Cripto-1 (CR-1) is a glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein that has been shown to play an important role in embryogenesis and cellular transformation. CR-1 is reported to function as a membrane-bound co-receptor and as a soluble ligand. Although a number of studies implicate the role of CR-1 as a soluble ligand in tumor progression, it is unclear how transition from the membrane-bound to the soluble form is physiologically regulated and whether differences in biological activity exist between these forms. Here, we demonstrate that CR-1 protein is secreted from tumor cells into the conditioned medium after treatment with serum, epidermal growth factor, or lysophosphatidic acid, and this soluble form of CR-1 exhibits the ability to promote endothelial cell migration as a paracrine chemoattractant. On the other hand, membranebound CR-1 can stimulate endothelial cell sprouting through direct cell-cell interaction. Shedding of CR-1 occurs at the GPI-anchorage site by the activity of GPI-phospholipase D (GPIPLD), because CR-1 shedding was suppressed by siRNA knockdown of GPI-PLD and enhanced by overexpression of GPI-PLD. These findings describe a novel molecular mechanism of CR-1 shedding, which may contribute to endothelial cell migration and possibly tumor angiogenesis.

    DOI: 10.1074/jbc.M702713200

    Web of Science

    researchmap

  • Development of bionanocapsules targeting brain tumors

    Yumi Tsutsui, Kazuhito Tomizawa, Mana Nagita, Hiroyuki Michiue, Tei-ichi Nishiki, Iori Ohmori, Masaharu Seno, Hideki Matsui

    JOURNAL OF CONTROLLED RELEASE   122 ( 2 )   159 - 164   2007.9

     More details

    Language:English   Publisher:ELSEVIER  

    Bionanocapsules (BNCs) are hollow nanoparticles that are composed of L protein (the hepatitis B virus surface antigen) and show specific affinity for human hepatocytes. The pre-S1 peptide displayed on the surface of BNCs is the specific ligand for binding to the receptor on human hepatocytes. Therefore, BNCs are not delivered to other tissues, such as the brain. The aim of the present study was to develop a novel drug delivery system (DIDS) targeting brain tumors using BNCs that selectively targeted brain tumors. Epidermal growth factor receptor (EGFR), especially a constitutively active genomic sequence deletion variant of EGFR (EGFRvIII), is overexpressed in human glioblastoma. In the present study, we replaced the pre-S1 peptide with the antibody affinity motif of protein A and made hybrid BNCs conjugated with anti-human EGFR antibody recognizing EGFRvIII. The hybrid BNCs were efficiently delivered to glioma cells but not normal glial cells. Moreover, we confirmed the specific delivery of the hybrid BNCs to brain tumors in an in vivo brain tumor model. These results suggest that this new approach using BNCs is a promising system for brain tumor-targeted drug delivery. (C) 2007 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jconrel.2007.06.019

    Web of Science

    researchmap

  • 'Crystal lattice engineering,' an approach to engineer protein crystal contacts by creating intermolecular symmetry: Crystallization and structure determination of a mutant human RNase 1 with a hydrophobic interface of leucines

    Hidenori Yamada, Taro Tamada, Megumi Kosaka, Kohei Miyata, Shinya Fujiki, Masaru Tano, Masayuki Moriya, Mamoru Yamanishi, Eijiro Honjo, Hiroko Tada, Takeshi Ino, Hiroshi Yamaguchi, Junichiro Futami, Masaharu Seno, Takashi Nomoto, Tomoko Hirata, Motonobu Yoshimura, Ryota Kuroki

    PROTEIN SCIENCE   16 ( 7 )   1389 - 1397   2007.7

     More details

    Language:English   Publisher:COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT  

    A protein crystal lattice consists of surface contact regions, where the interactions of specific groups play a key role in stabilizing the regular arrangement of the protein molecules. In an attempt to control protein incorporation in a crystal lattice, a leucine zipper-like hydrophobic interface (comprising four leucine residues) was introduced into a helical region (helix 2) of the human pancreatic ribonuclease 1 (RNase 1) that was predicted to form a suitable crystallization interface. Although crystallization of wild-type RNase 1 has not yet been reported, the RNase 1 mutant having four leucines (4L-RNase 1) was successfully crystallized under several different conditions. The crystal structures were subsequently determined by Xray crystallography by molecular replacement using the structure of bovine RNase A. The overall structure of 4L-RNase 1 is quite similar to that of the bovine RNase A, and the introduced leucine residues formed the designed crystal interface. To characterize the role of the introduced leucine residues in crystallization of RNase 1 further, the number of leucines was reduced to three or two (3L-and 2L-RNase 1, respectively). Both mutants crystallized and a similar hydrophobic interface as in 4L-RNase 1 was observed. A related approach to engineer crystal contacts at helix 3 of RNase 1 (N4L- RNase 1) was also evaluated. N4L- RNase 1 also successfully crystallized and formed the expected hydrophobic packing interface. These results suggest that appropriate introduction of a leucine zipper-like hydrophobic interface can promote intermolecular symmetry for more efficient protein crystallization in crystal lattice engineering efforts.

    DOI: 10.1110/ps.072851407

    Web of Science

    researchmap

  • Characterization of bio-nanocapsule as a transfer vector targeting human hepatocyte carcinoma by disulfide linkage modification

    Tadahiro Nagaoka, Takayuki Fukuda, Shinnosuke Yoshida, Hirohito Nishimura, Dongwei Yu, Shun'ichi Kuroda, Katsuyuki Tanizawa, Akhko Kondo, Masakazu Ueda, Hidenori Yamada, Hiroko Tada, Masaharu Seno

    JOURNAL OF CONTROLLED RELEASE   118 ( 3 )   348 - 356   2007.4

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    The bio-nanocapsules (BNCs) composed of the recombinant envelope L-protein of hepatitis B virus constitute efficient delivery vectors specifically targeting human hepatocytes. Here, we have tried to enhance the stability of the BNCs because the L-proteins in the BNCs were aggregated due to random disulfide bridging when stored for a long period at 4 degrees C. The envelope protein contains fourteen cysteine residues in the S domain. Aggregation of the envelope proteins might be avoided if unessential cysteine residues are replaced or removed because the irreversible alkylation of the free sulfhydryl group protects against the aggregation and enhances the efficiency of encapsulation. In this study, the possibility of reducing the number of cysteine residues in the S domain to enhance the stability of the BNCs was assessed. The replacement of each cysteine residue by site-directed mutation showed that nine of fourteen cysteine residues were not essential to obtaining BNCs secreted into the culture media. Furthermore, upon evaluating the combination of these mutations, it was found that eight residues of replacement were acceptable. The mutant BNCs with replaced eight cysteine residues were not only more resistant against trypsin, but also more effective in transducing genes into human hepatoma-derived HepG2 cells than the original type BNC. Thus, we demonstrated that the minimized number of cysteine residues in the S domain could enhance the stability of the BNCs. (c) 2006 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jconrel.2006.12.020

    Web of Science

    researchmap

  • AR42J細胞の分化に対する細胞外基質の影響

    濱本 耕平, 山田 聡子, 山本 頼綱, 小寺 力, 原 朱美, 妹尾 昌治, 小島 至

    糖尿病   50 ( Suppl.1 )   S - 349   2007.4

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

    researchmap

  • Gene therapy of liver tumors with human liver-specific nanoparticles (vol 14, pg 74, 2007)

    M. Ueda, Y. Iwasaki, T. Yamada, A. Kondo, M. Seno, K. Tanizawa, S. Kuroda, M. Sakamoto, M. Kitajima

    CANCER GENE THERAPY   14 ( 4 )   440 - 440   2007.4

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    DOI: 10.1038/sj.cgt.7701031

    Web of Science

    researchmap

  • Characteristics of hollow microtubes consisting of amorphous iron oxide nanoparticles produced by iron oxidizing bacteria, Leptothrix ochracea

    H. Hashimoto, S. Yokoyama, H. Asaoka, Y. Kusano, Y. Ikeda, M. Seno, J. Takada, T. Fujii, M. Nakanishi, R. Murakami

    JOURNAL OF MAGNETISM AND MAGNETIC MATERIALS   310 ( 2 )   2405 - 2407   2007.3

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    Some features of characteristic iron oxide sheaths which the iron oxidizing bacteria Leptothrix ochracea ( L. oceracea) formed were studied in order to make clear their morphology microstructure, chemical composition, and crystal structure through scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDX), and X-ray diffraction (XRD). Each sheath was a hollow tube with average outer and inner diameters of 1.1 and 1.4 m, respectively. Their length ranged from 10 to 200 mu m and the aspect ratio was 10-200. Each sheath was constructed by very small particles with a diameter of less than 100 nm. The hollow sheaths were mainly composed of Fe and O with small amounts of Si and P. The chemical composition analyzed by EDX was roughly Fe: Si: P = 80: 15: 5 with the exception of O. XRD measurement revealed that crystal structures of the sheath were similar to that of 2-line ferrihydrite. The sheath showed spin-glass-like magnetic properties. (c) 2006 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jmmm.2006.10.793

    Web of Science

    researchmap

  • 脳腫瘍を標的としたバイオナノカプセルの開発

    筒井 佑美, 富澤 一仁, 西木 禎一, 大守 伊織, 名木田 真奈, 妹尾 昌治, 松井 秀樹

    日本生理学雑誌   69 ( 3 )   125 - 125   2007.3

     More details

    Language:Japanese   Publisher:(一社)日本生理学会  

    researchmap

  • Gene therapy of liver tumors with human liver-specific nanoparticles

    Y. Iwasaki, M. Ueda, T. Yamada, A. Kondo, M. Seno, K. Tanizawa, S. Kuroda, M. Sakamoto, M. Kitajima

    CANCER GENE THERAPY   14 ( 1 )   74 - 81   2007.1

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    The development of safe and efficient liver-specific gene delivery approaches offers new perspectives for the treatment of liver disease, in particular, liver cancer. We evaluated the therapeutic potential of hepatotropic nanoparticles for gene therapy of liver tumor. These nanoparticles do not contain a viral genome and display the hepatitis B virus L antigen, which is essential to confer hepatic specificity. It has not been shown whether a therapeutic effect could be obtained using L nanoparticles in a human liver tumor xenograft model. Rats bearing human hepatic ( NuE) and non-hepatic tumors were injected with L nanoparticles containing a green fluorescent protein ( GFP) expression plasmid. GFP expression was observed only in NuE-derived tumors but not in the non-hepatic tumor. The potential for treatment of liver tumors was analyzed using L nanoparticles containing the herpes simplex virus thymidine kinase gene, in conjunction with ganciclovir pro-drug administration. The growth of NuE-derived tumors in L particle-injected rats was significantly suppressed, but not of the non-hepatic tumor control. In summary, this is the first demonstration that nanoparticles could be used for delivery of therapeutic genes with anti-tumor activity into human liver tumors. This intravenous delivery system may be one of the major advantages as compared to many other viral vector systems.

    DOI: 10.1038/sj.cgt.7700990

    Web of Science

    researchmap

  • 心筋症治療薬の開発シーズ:好酸球由来塩基性タンパク質

    妹尾昌治

    ちゅうごく産業創造センター会報   No.74, p40-41.   2007

     More details

  • バイオナノキャリアの開発とがん遺伝子治療への応用

    近藤昭彦, 黒田俊一, 谷澤克行, 妹尾昌治, 上田政和

    バイオテクノロジージャーナル   7 ( 1 )   41 - 47   2007

     More details

  • Conophylline and betacellulin delta 4 promote differentiation of pancreatic cells both in vitro and in vivo

    Tsutomu Kodera, Takeki Ogata, Yoritsuna Yamamoto, Kazuo Umezawa, Masaharu Seno, Yuji Tanaka, Itaru Kojima, Yasuhiro Watanabe

    JOURNAL OF PHARMACOLOGICAL SCIENCES   103   80P - 80P   2007

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:JAPANESE PHARMACOLOGICAL SOC  

    Web of Science

    researchmap

  • Anti-tumor effect in an in vivo model by human-derived pancreatic RNase with basic fibroblast growth factor insertional fusion protein through antiangiogenic properties

    Hiroshi Yagi, Masakazu Ueda, Hiromitsu Jinno, Koichi Aiura, Shuji Mikami, Hiroko Tada, Masaharu Seno, Hidenori Yamada, Masaki Kitajima

    CANCER SCIENCE   97 ( 12 )   1315 - 1320   2006.12

     More details

    Language:English   Publisher:BLACKWELL PUBLISHING  

    It is thought that the export of angiogenic fibroblast growth factors (FGF) from tumors may be involved in the onset of tumor angiogenesis. To create a new active targeting drug that inhibits the tumor angiogenic process without toxicities to normal cells, human basic FGF (h-bFGF) was inserted genetically into the Gly89 position of cross-linked RNase1 (the ribonuclease inhibitor protein [RI] binding site of cross-linked human pancreatic RNase) to prevent stereospecific binding to RI. The resultant insertional-fusion protein (CL-RFN89) was active both as h-bFGF and as RNase1. Furthermore, it acquired an additional ability of evading RI through steric blockade of RI binding caused by the fused h-bFGF domain. In the present study, the effect of the resultant protein, CL-RFN89, on the antitumor response though its antiangiogenic properties was investigated in an in vivo model. Continuous systemic treatment with CL-RFN89 significantly inhibited the growth of human A431 squamous cell carcinomas in vivo. Seven days of treatment with CL-RFN89 resulted in a 58.2% inhibition of tumor growth compared with control mice (P &lt; 0.0001). Furthermore, immunohistochemistry using a rat antimouse CD31 antibody showed that treatment with CL-RFN89 reduced tumor vascularization. These findings identify CL-RFN89 as a potent systemic inhibitor of tumor growth as a result of its antiangiogenic properties. This protein appears to be a new systemic antitumor agent.

    DOI: 10.1111/j.1349-7006.2006.00336.x

    Web of Science

    researchmap

  • Secretory production system of bionanocapsules using a stably transfected insect cell line

    Takuya Shishido, Masaru Muraoka, Masakazu Ueda, Masaharu Seno, Katsuyuki Tanizawa, Shun'ichi Kuroda, Hideki Fukuda, Akihiko Kondo

    APPLIED MICROBIOLOGY AND BIOTECHNOLOGY   73 ( 3 )   505 - 511   2006.12

     More details

    Language:English   Publisher:SPRINGER  

    Bionanocapsules (BNCs) are hollow nanoscale particles composed of L protein of the hepatitis B virus surface antigen that represent specific affinity for human hepatocytes. BNCs can transfer genes and drugs into human hepatocytes efficiently and specifically. BNC can be expressed in yeast cells. In this study, we developed a new L particle production system using a stably transfected insect cell line. For this purpose, we established a host vector system using the Trichoplusia ni insect cell line. L particles were efficiently secreted by the overexpression of the L protein, which was fused to the secretion signal peptide. The concentration of L particles was reached approximately 1.7 mu g/ml in 5 days during cultivation in a serum-free medium without antibiotic selective pressure. The production of L particles was maintained for at least 75 days. The secretory production of L particles facilitated their easy purification by chromatography. Furthermore, it was demonstrated that purified L particles can transfect only human hepatocytes. Therefore, an insect cell expression system is an attractive tool for the production of BNC.

    DOI: 10.1007/s00253-006-0486-3

    Web of Science

    researchmap

  • Hydrogen peroxide and endothelin-1 are novel activators of betacellulin ectodomain shedding

    Michael P. Sanderson, Catherine A. Abbott, Hiroko Tada, Masaharu Seno, Peter J. Dempsey, Andrew J. Dunbar

    JOURNAL OF CELLULAR BIOCHEMISTRY   99 ( 2 )   609 - 623   2006.10

     More details

    Language:English   Publisher:WILEY-LISS  

    The betacellulin precursor (pro-BTC) is a novel substrate for ADAM10-mediated ectodomain shedding. In this report, we investigated the ability of novel physiologically relevant stimuli, including G-protein coupled receptor (GPCR) agonists and reactive oxygen species (ROS), to stimulate pro-BTC shedding. We found that in breast adenocarcinoma MCF7 cells overexpressing pro-BTC, hydrogen peroxide (H2O2) was a powerful stimulator of ectodomain shedding. The stimulation of pro-BTC shedding by H2O2 was blocked by the broad-spectrum metalloprotease inhibitor TAPI-0 but was still functional in ADAM17 (TACE)-deficient stomach epithelial cells indicating the involvement of a distinct metalloprotease. H2O2-induced pro-BTC shedding was blocked by co-culturing cells in the anti-oxidant N-acetyl-L-cysteine but was unaffected by culture in calcium-deficient media. By contrast, calcium ionophore, which is a previously characterized activator of pro-BTC shedding, was sensitive to calcium depletion but was unaffected by co-culture with the anti-oxidant, identifying a clear distinction between these stimuli. We found that in vascular smooth muscle cells overexpressing pro-BTC, the GPCR agonist endothelin-1 (ET-1) was a strong inducer of ectodomain shedding. This was blocked by a metalloprotease inhibitor and by overexpression of catalytically inactive E385AADAM10. However, overexpression of wild-type ADAM10 or ADAM17 led to an increase in ET-1-induced pro-BTC shedding providing evidence for an involvement of both enzymes in this process. This study identifies ROS and ET-1 as two novel inducers of pro-BTC shedding and lends support to the notion of activated shedding occurring under the control of physiologically relevant stimuli. J. Cell. Biochem. 99: 609-623, 2006. (c) 2006 Wiley-Liss, Inc.

    DOI: 10.1002/jcb.20968

    Web of Science

    researchmap

  • Denatured and reversibly cationized p53 readily enters cells and simultaneously folds to the functional protein in the cells

    H Murata, M Sakaguchi, J Futami, M Kitazoe, T Maeda, H Doura, M Kosaka, H Tada, M Seno, N Huh, H Yamada

    BIOCHEMISTRY   45 ( 19 )   6124 - 6132   2006.5

     More details

    Language:English   Publisher:AMER CHEMICAL SOC  

    Cationization is a powerful strategy for internalizing a protein into living cells. On the other hand, a reversibly cationized denatured protein through disulfide bonds is not only soluble in water but also able to fold to the native conformation in vitro. When these advantages in cationization were combined, we developed a novel method to deliver a denatured protein into cells and simultaneously let it fold to express its function within cells. This "in-cell folding" method enhances the utility of recombinant proteins expressed in Escherichia coli as inclusion bodies; that is, the recombinant proteins in inclusion bodies are solubilized by reversible cationization through cysteine residues by disulfide bonds with aminopropyl methanethiosulfonate or pyridyldithiopropionylpolyethylenimine and then incubated with cells without an in vitro folding procedure. As a model protein, we investigated human tumor-suppressor p53. Treatment of p53-null Saos-2 cells with reversibly cationized p53 revealed that all events examined as indications of the activation of p53 in cells, such as reduction of disulfide bonds followed by tetramer formation, localization into the nucleus, induction of p53 target genes, and induction of apoptosis of cells, occurred. These results suggest that reversible cationization of a denatured protein through cysteine residues is an alternative method for delivery of a functional protein into cells. This method would be very useful when a native folded protein is not readily available.

    DOI: 10.1021/bi052642a

    Web of Science

    researchmap

  • Mechanisms of the growth-inhibitory effect of the RNase-EGF fused protein against EGFR-overexpressing cells

    S Hoshimoto, M Ueda, H Jinno, M Kitajima, J Futami, M Seno

    ANTICANCER RESEARCH   26 ( 2A )   857 - 863   2006.3

     More details

    Language:English   Publisher:INT INST ANTICANCER RESEARCH  

    Background: We previously showed the usefulness of a fused protein of human pancreatic ribonuclease1 (hRNase1) with human epidermal growth factor (hEGF) for molecular targeting of EGF receptor (EGFR)-overexpressing cells. In this study, the mechanisms underlying the inhibition of cell growth by RNase-EGF fused proteins was confirmed. Materials and Methods: Des. 1-7 hRNase1 was genetically fused to hEGF. The fused proteins were expressed and isolated from Escherichia coli. The internalization of hRNase1-hEGF was confirmed by confocal fluorescence microscopy. The growth-inhibitory effect of the fused proteins was evaluated by MTT assay. Results: FITC-labelled hRNase1-hEGF was internalized into EGFR-overexpressing A431 cells. The internalization was not observed in A 431 cells pre-treated with hEGF and EGFR-deficient H69 cells. The growth-inhibitory effect of des.1-7 hRNase1-hEGF against A431 cells was statistically significantly more pronounced than that of hRNase1-hEGF. Conclusion: RNase-EGF fused proteins are internalized through EGFR and inhibit cell growth by exerting their ribonucleolytic activity in the cytosol.

    Web of Science

    researchmap

  • Engineered bio-nanocapsules, the selective vector for drug delivery system

    DW Yu, T Fukuda, Tuoya, S Kuroda, K Tanizawa, A Kondo, M Ueda, T Yamada, H Tada, M Seno

    IUBMB LIFE   58 ( 1 )   1 - 6   2006.1

     More details

    Language:English   Publishing type:Book review, literature introduction, etc.   Publisher:TAYLOR & FRANCIS INC  

    The bio-nanocapsule (BNC) is our concept of artificial hollow nanoparticles that have been designed and produced through biotechnological procedures. We proposed an empty virus-like particle, which consists of a recombinant L envelope protein of hepatitis B virus (HBV) and a lipid derived from the host cell, as an engineered BNC. Although this BNC was first developed as an immunogen of hepatitis B vaccine, the pre-S1 region in N-terminus of L envelope protein confers hepatocyte specific infectivity of HBV on the BNC. This recombinant BNC is now being developed as a novel platform of drug delivery system (DDS) vector for selective delivery.

    DOI: 10.1080/15216540500484368

    Web of Science

    researchmap

  • Development of dds using hollow bio-nanoparticles and their commercialization

    Akihiko Kondo, Shun-ichi Kuroda, Katsuyuki Tanizawa, Masaharu Seno, Masakazu Ueda

    Drug Delivery System   21 ( 4 )   435 - 443   2006

     More details

    Language:English  

    We succeeded overproduction of the HBV envelope L particles with an approximate average particle size of 80 nm in yeast cells. Because the L particle is an empty bionanoparticles containing no viral DNA, it can be used as a safe and efficient carrier for human liver-specific delivery (pinpoint delivery) of drug and gene. In addition, genetically engineered L particles that are able to target to various organs were constructed by deleting the hepatocyte binding domain of L protein (pre-S region) and displaying targeting peptide or protein ligands. Therefore, bionanoparticles are a novel nano-carrier applicable to the broad range of pinpoint DDS. © 2006, THE JAPAN SOCIETY OF DRUG DELIVERY SYSTEM. All rights reserved.

    DOI: 10.2745/dds.21.435

    Scopus

    researchmap

  • Separation of Dual Affinity of Betacellulin to ErbB Receptors

    T. Nagaoka, H. Tada, H. Yamada, M. Seno

    MOLECULAR BIOLOGY OF THE CELL   17   2006

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Betacellulin-delta 4, a novel differentiation factor for pancreatic beta-cells, ameliorates glucose intolerance in streptozotocin-treated rats

    T Ogata, AJ Dunbar, Y Yamamoto, Y Tanaka, M Seno, Kojima, I

    ENDOCRINOLOGY   146 ( 11 )   4673 - 4681   2005.11

     More details

    Language:English   Publisher:ENDOCRINE SOC  

    We previously described a novel alternatively spliced mRNA transcript of the betacellulin (BTC) gene. This splice isoform, termed BTC-delta 4, lacks the C-loop of the epidermal growth factor motif and the transmembrane domain as a result of exon 4 'skipping'. In this study, we expressed BTC-delta 4 recombinantly to explore its biological function. When BTC-delta 4 was expressed in COS-7 cells, it was secreted largely into the culture medium, in contrast to BTC. Unlike BTC, highly purified recombinant BTC-delta 4 produced in Escherichia coli failed to bind or induce tyrosine phosphorylation of either ErbB1 or ErbB4, nor did it antagonize the binding of BTC to these receptors. Consistent with this, BTC-delta 4 failed to stimulate DNA synthesis in Balb/c 3T3 and INS-1 cells. However, BTC-delta 4 induced differentiation of pancreatic beta-cells; BTC-delta 4 converted AR42J cells to insulin-producing cells. When recombinant BTC-delta 4 was administered to streptozotocin-treated neonatal rats, it reduced the plasma glucose concentration and improved glucose tolerance. Importantly, BTC-delta 4 significantly increased the insulin content, the beta-cell mass, and the numbers of islet-like cell clusters and PDX-1-positive ductal cells. Thus, BTC-delta 4 is a secreted protein that stimulates differentiation of beta-cells in vitro and in vivo in an apparent ErbB1- and ErbB4- independent manner. The mechanism by which BTC-delta 4 exerts this action on beta-cells remains to be defined but presumably involves an, as yet, unidentified unique receptor.

    DOI: 10.1210/en.2005-0456

    Web of Science

    researchmap

  • Identification of cell surface marker candidates on SV-T2 cells using DNA microarray on DLC-coated glass

    Tuoya, K Hirayama, T Nagaoka, DW Yu, T Fukuda, H Tada, H Yamada, M Seno

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   334 ( 1 )   263 - 268   2005.8

     More details

    Language:English   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We analyzed gene expression profiles of normal mouse fibroblast BALB/c 3T3 cells and its SV40 transformant SV-T2 cells using our originally developed cell surface marker DNA microarray, which is prepared on a diamond-like carbon-coated glass. As a result, CD62L and IL-6 receptor alpha gene expressions were upregulated in SV-T2 and were thought to be candidates for cell surface markers of the cells. The result of microarray analysis was validated by real-time quantitative PCR, immunohistochemistry and biological assays. These data show that our cell surface marker DNA microarray should be useful in finding the candidates of cell type-specific surface markers. (c) 2005 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2005.06.083

    Web of Science

    researchmap

  • 細胞および組織特異的遺伝子導入を可能にするバイオナノカプセル

    YAMADA Tadanori, SENO Masaharu, UEDA Masakazu, KONDO Akihiko, TANIZAWA Katsuyuki, KURODA Shun'ichi

    生物工学会誌   83・8・380-383   2005.8

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

    researchmap

  • Anti-angiogenic effect of an insertional fusion protein of human basic fibroblast growth factor and ribonuclease-1

    T Hayashida, M Ueda, K Aiura, H Tada, M Onizuka, M Seno, H Yamada, M Kitajima

    PROTEIN ENGINEERING DESIGN & SELECTION   18 ( 7 )   321 - 327   2005.7

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    Human pancreatic ribonuclease-1 (RNase1) does not exhibit its cytotoxicity unless it is artificially internalized into the cytosol. Furthermore, once it encounters the cytosolic RNase inhibitor (RI), the activity of RNase1 is seriously reduced. To achieve the cellular targeting of RNase1 and the blocking of RI binding simultaneously, the basic fibroblast growth factor (bFGF) sequence was inserted into RNase1 at the RI binding site using a gene fusion technique. The effect of this fusion protein, CL-RFN89, on the angiogenesis, which was accelerated by FGF-FGF receptor interaction, was investigated. It was shown by using fluorescein-labeled CL-RFN89, that the binding to human umbilical vein endothelial cells (HUVECs) was dependent on the existence of the FGF receptors. In addition, CL-RFN89 inhibited the cellular growth of HUVECs in vitro and also inhibited the tube formation, using a three-dimensional tube formation assay. Furthermore, this fusion protein was shown to prevent in vivo tumor cell-induced angiogenesis, using the mouse dorsal air sac assay. These results demonstrated that CL-RFN89 inhibits angiogenesis in vitro and in vivo and that it can be expected to be a potent antiangiogenic agent.

    DOI: 10.1093/protein/gzi040

    Web of Science

    researchmap

  • The specific delivery of proteins to human liver cells by engineered bio-nanocapsules

    DW Yu, C Amano, T Fukuda, T Yamada, S Kuroda, K Tanizawa, A Kondo, M Ueda, H Yamada, H Tada, M Seno

    FEBS JOURNAL   272 ( 14 )   3651 - 3660   2005.7

     More details

    Language:English   Publisher:BLACKWELL PUBLISHING  

    A bio-nanocapsule (BNC), composed of the surface antigen (sAg) of the hepatitis B virus, is an efficient nanomachine with which to accomplish the liver-specific delivery of genes and drugs. Approximately 110 molecules of sAg are associated to form a BNC particle with an average diameter of 130 nm. The L protein is an sAg peptide composed mainly of preS and S regions. The preS region, with specific affinity for human hepatocytes, is localized in the N-terminus. The S region following the preS has two transmembrane regions responsible for the formation of particles. In this study, the fusion of emerald green fluorescent protein (EGFP) at the C-terminus of the S region was designed to deliver proteins to human hepatocytes. Truncation of the C-terminus of the S region was required to obtain sufficient expression levels in Cos7 cells. The nanoparticles that were produced delivered EGFP to human hepatoma cells, displaying the EGFP moiety outside, or enclosing it inside. However, only a single orientation characterizes the particle, so that either type of L fusion particle could be effectively and independently separated by an antibody affinity column. The dual C-terminal topologies of the L fusion particles designed in this study could be applied to various proteins for the C-terminal moiety of the L fusion proteins, depending on the character of the proteins, such as cytoplasmic proteins, as well as cytokines or ligands to cell surface receptors. We suggest that this fusion design is the most efficient way to prepare a BNC that delivers proteins to specific cells or tissues.

    DOI: 10.1111/j.1742-4658.2005.04790.x

    Web of Science

    researchmap

  • Protein transduction assisted by polyethylenimine-cationized carrier proteins

    M Kitazoe, H Murata, J Futami, T Maeda, M Sakaguchi, M Miyazaki, M Kosaka, H Tada, M Seno, N Huh, M Namba, M Nishikawa, Y Maeda, H Yamada

    JOURNAL OF BIOCHEMISTRY   137 ( 6 )   693 - 701   2005.6

     More details

    Language:English   Publisher:JAPANESE BIOCHEMICAL SOC  

    Previously, we have reported that cationized-proteins covalently modified with polyethylenimine (PEI) (direct PEI-cationization) efficiently enter cells and function in the cytosol [Futami et al. (2005) J. Biosci. Bioeng. 99,95-1031. However, it may be more convenient if a protein could be delivered into cells just by mixing the protein with a PEI-cationized carrier protein having a specific affinity (indirect PEI-cationization). Thus, we prepared PEI-cationized avidin (PEI-avidin), streptavidin (PEI-streptavidin), and protein G (PEI-protein G), and examined whether they could deliver biotinylated proteins and antibodies into living cells. PEI-avidin (and/or PEI-streptavidin) carried biotinylated GFPs into various mammalian cells very efficiently. A GFP variant containing a nuclear localization signal was found to arrive even in the nucleus. The addition of a biotinylated RNase A derivative mixed with PEI-streptavidin to a culture medium of 3T3-SV-40 cells resulted in remarkable cell growth inhibition, suggesting that the biotinylated RNase A derivative entered cells and digested intracellular RNA molecules. Furthermore, the addition of a fluorescein-labeled antiS100C (beta-actin binding protein) antibody mixed with PEI-protein G to human fibroblasts resulted in the appearance of a fluorescence image of actin-like filamentous structures in the cells. These results indicate that indirect PEI-cationization using non-covalent interaction is as effective as the direct PEI-cationization for the transduction of proteins into living cells and for expression of their functions in the cytosol. Thus, PEI-cationized proteins having a specific affinity for certain molecules such as PEI-streptavidin, PEI-avidin and PEI-protein G are concluded to be widely applicable protein transduction carrier molecules.

    DOI: 10.1093/jb/mvi081

    Web of Science

    researchmap

  • Conophylline and betacellulin-delta 4: An effective combination to induce differentiation of pancreatic beta cells

    RI Kitamura, T Ogata, Y Tanaka, MH Seno, Takei, I, K Umezawa, Kojima, I

    DIABETES   54 ( 2 )   A393 - A393   2005

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER DIABETES ASSOC  

    DOI: 10.1507/endocrj.K06-199

    Web of Science

    researchmap

  • Intracellular delivery of proteins into mammalian living cells by polyethylenimine-cationization

    Junichiro Futami, Midori Kitazoe, Takashi Maeda, Emiko Nukui, Masakiyo Sakaguchi, Jun Kosaka, Masahiro Miyazaki, Megumi Kosaka, Hiroko Tada, Masaharu Seno, Junzo Sasaki, Nam-Hu Huh, Masayoshi Namba, Hidenori Yamada

    Journal of Bioscience and Bioengineering   99 ( 2 )   95 - 103   2005

     More details

    Language:English   Publisher:Society of Fermentation and Bioengineering  

    In the post-genomic era, there is pressing need for development of protein manipulation methodology to analyze functions of proteins in living cells. For this purpose, techniques to deliver functional proteins into living cells are currently being evaluated as alternative approaches to the introduction of transcriptionally active DNA. Here, we describe a novel method for efficient protein transduction into living cells in which a protein is simply cationized with polyethylenimine (PEI) by limited chemical conjugation. PEI-cationized proteins appear to adhere to the cell surface by ionic charge interaction and then internalize into cells in a receptor- and transporter-independent fashion. Since PEI is an organic macromolecule with a high cationic-charge density, limited coupling with PEI results in endowment of sufficient cationic charge to proteins without causing serious decline in their fundamental functions. A number of PEI-cationized proteins, such as ribonuclease (RNase), green fluorescent protein (GFP) and immunoglobulin (IgG), efficiently entered cells and functioned in the cytosol. Our results suggest that protein cationization techniques using PEI will be useful for the development of protein transduction technology. © 2005, The Society for Biotechnology.

    DOI: 10.1263/jbb.99.95

    Scopus

    PubMed

    researchmap

  • Differentiation of adult hepatic stem-like cells into pancreatic endocrine cells

    S Yamada, K Terada, Y Ueno, T Sugiyama, M Seno, Kojima, I

    CELL TRANSPLANTATION   14 ( 9 )   647 - 653   2005

     More details

    Language:English   Publisher:COGNIZANT COMMUNICATION CORP  

    To apply cell transplantation for treatment of diabetes mellitus, a sufficient number of beta-cell sources are required. In the present study, we examined whether an epithelial cell line obtained from normal adult rat liver, namely hepatic stem-like (HSL) cells, which can be converted to both hepatocytes and billiary epithelial cells, could be a potential beta-cell source. The growth speed of HSL cells was rapid and these cells were easily expanded in vitro. Bipotential hepatic stem cells, HSL cells, also expressed PGP9.5, which is expressed in neurons, beta-cells, and progenitor cells of the pancreatic endocrine cells as well. Sodium butyrate induced morphological changes in HSL cells and converted them into flattened cells with large cytoplasm. When HSL cells were incubated with a combination of 5 mM sodium butyrate and I nM betacellulin, most of the cells were converted into morphologically neuron-like cells. RT-PCR analysis revealed that a series of transcriptional factors involved in differentiation of pancreatic endocrine cells was induced by the treatment with sodium butyrate and betacellulin. mRNAs for insulin, pancreatic polypeptide, and somatostatin were also observed. Immunoreactive pancreatic polypeptide, somatostatin, and insulin were detected in sodium butyrate and betacellulin-treated HSL cells. In conclusion, HSL cells obtained from adult normal liver also have the potential to differentiate into pancreatic endocrine cells in vitro. HSL cells may be one of the potential beta-cell sources for cell transplant therapy for insulin-dependent diabetes.

    DOI: 10.1016/j.ssc.2005.01.003

    Web of Science

    researchmap

  • 革新的なナノキャリア:中空バイオナノ粒子によるピンポイントDDS

    近藤昭彦, 黒田俊一, 谷澤克行, 妹尾昌治, 上田政和

    バイオインダストリー   22 ( 5 )   22 - 27   2005

     More details

  • 中空バイオナノ粒子を用いたピンポイントDDSの開発

    近藤昭彦, 黒田俊一, 谷澤克行, 妹尾昌治

    Chemical Engineering   50 ( 5 )   351 - 356   2005

     More details

  • Epithelial mesenchymal transition is a characteristic of hyperplasias and tumors in mammary gland from MMTV-cripto-1 transgenic mice

    L Strizzi, C Bianco, N Normanno, M Seno, C Wechselberger, B Wallace-Jones, NI Khan, M Hirota, YP Sun, M Sanicola, DS Salomon

    JOURNAL OF CELLULAR PHYSIOLOGY   201 ( 2 )   266 - 276   2004.11

     More details

    Language:English   Publisher:WILEY-LISS  

    Epithelial-mesenchymal transition (EMT) facilitates migration and invasion of epithelial tumor cells. Cripto-1 (CR-1), a member of the epidermal growth factor-CFC protein family increases migration of cells in vitro. Here the expression of molecular markers and signaling molecules characteristic of EMT were assessed in mammary gland hyperplasias and tumors from mice expressing the human CR-1 transgene by the MMTV promoter (MMTV-CR-1) and in mouse mammary epithelial cell line HC-11 overexpressing CR-1 (HC-11/CR-1). Western blot analysis showed decreased expression of E-cadherin in MMTV-CR-1 tumors and in HC-11/CR-1 cells. The expression of N-cadherin, vimentin, cyclin-D1, and of the zinc-finger transcription factor, snail, was increased in MMTV-CR-1 tumors. Increased snail mRNA was also found in HC-11/CR-1 cells. Expression of phosphorylated (N-c-Src, P-focal adhesion kinase (FAK), P-Akt, P-glycogen synthease kinase 30 (GSK-3beta), dephosphorylated (DP)-beta-catenin, and various integrins such as, alpha 3, alpha v, beta 1, beta 3, and beta 4 was also increased in MMTV-CR-1 tumors. Immunohistochemistry showed positive staining for vimentin, N-cadherin, cyclin-D1, smooth muscle actin, fibronectin, snail, and beta-catenin in MMTV-CR-1 tumor sections. HC-11/CR-1 cells treated with the c-Src inhibitor PP2 reduced the expression of P-c-Src and of P-FAK, P-Akt, P-GSK-3beta, DP-beta-catenin all known to be activated by c-Src. Migration of HC-1I/CR-1 cells was also reduced by PP2 treatment. These results suggest that CR-1 may play a significant role in promoting the increased expression of markers and signaling molecules associated with EMT. (C) 2004 Wiley-Liss, Inc.

    DOI: 10.1002/jcp.20062

    Web of Science

    researchmap

  • Characterization of cell surface marker proteins in SV-T2 cells

    Y Tuo, K Hirayama, T Nagaoka, H Tada, H Yamada, M Seno

    MOLECULAR BIOLOGY OF THE CELL   15   324A - 325A   2004.11

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction

    H Tada, M Onizuka, K Muraki, W Masuzawa, J Futami, M Kosaka, M Seno, H Yamada

    FEBS LETTERS   568 ( 1-3 )   39 - 43   2004.6

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    Basic fibroblast growth factor (bFGF) was inserted in the middle of human ribonuclease 1 (RNase1) sequence at an RNase inhibitor (RI)-binding site (Gly89) by a new gene fusion technique, insertional-fusion. The resultant insertional-fusion protein (CL-RFN89) was active both as bFGF and as RNase. Furthermore, it acquired an additional ability of evading RI through steric blockade of RI-binding caused by fused bFGF domain. As a result, CL-RFN89 showed stronger growth inhibition on B16/BL6 melanoma cells than an RI-sensitive tandem fusion protein. Thus, the insertional-fusion technique increases accessible positions for gene fusion on RNase, resulting in construction of a potent cytotoxic RNase. (C) 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.febslet.2004.05.007

    Web of Science

    researchmap

  • Reversal of streptozotocin-incluced hyperglycemia by transplantation of pseudoislets consisting of beta cells derived from ductal cells

    T Ogata, KY Park, M Seno, Kojima, I

    ENDOCRINE JOURNAL   51 ( 3 )   381 - 386   2004.6

     More details

    Language:English   Publisher:JAPAN ENDOCRINE SOCIETY  

    The present study was conducted in an attempt to treat streptozotocin (STZ)-induced hyperglycemia by transplanting beta cells derived from pancreatic ductal cells. Ductal cells obtained from neonatal rats were cultured in vitro. Approximately 70% of the cells were converted to insulin-secreting cells by incubating with betacellulin and activin A. Differentiated cells responded to a depolarizing concentration of potassium, tolbutamide and a high concentration of glucose, and insulin secretion increased by 2.5-, 2.3- and 1.6-fotd, respectively. We then prepared pseudoislets using the differentiated cells, which exhibited greatly improved glucose-responsiveness, with a high concentration of glucose inducing a 3-fold increase in insulin secretion. We transplanted these pseudoislets into the portal vein of STZ-treated nude mice. Before transplantation, the plasma glucose concentration was above 400 mg/dl, and after transplantation it was markedly reduced, the effect of which persisted for two weeks. These results indicate that STZ-induced hyperglycemia can be treated by transplanting pseudoislets consisting of 0 cells derived from ductal cells.

    DOI: 10.1507/endocrj.51.381

    Web of Science

    researchmap

  • Activin A and betacellulin - Effect on regeneration of pancreatic beta-cells in neonatal streptozotocin-treated rats

    L Li, ZH Yi, M Seno, Kojima, I

    DIABETES   53 ( 3 )   608 - 615   2004.3

     More details

    Language:English   Publisher:AMER DIABETES ASSOC  

    Activin A and betacellulin (BTC) are thought to regulate differentiation of pancreatic beta-cells during development and regeneration of beta-cells in adults. In the present study, we used neonatal rats treated with streptozotocin (STZ) to investigate the effects of activin A and BTC on regeneration of pancreatic beta-cells. One-dayold Sprague-Dawley rats were injected with STZ (85 mug/g) and then administered for 7 days with activin A and/or BTC. Treatment with activin A and BTC significantly reduced the plasma glucose concentration and the plasma glucose response to intraperitoneal glucose loading. The pancreatic insulin content and beta-cell mass in rats treated with activin A and BTC were significantly increased compared with the control group on day 8 and at 2 months. Treatment with activin A and BTC significantly increased the DNA synthesis in preexisting beta-cells, ductal cells, and 8-cells. The number of islet cell-like clusters (ICCs) and islets was significantly increased by treatment with activin A and BTC. In addition, the number of insulin/somatostatin-positive cells and pancreatic duodenal homeobox-1/somatostatin-positive cells was significantly increased. These results indicate that, in neonatal STZ-treated rats, a combination of activin A and BTC promoted regeneration of pancreatic beta-cells and improved glucose metabollism. in adults.

    DOI: 10.2337/diabetes.53.3.608

    Web of Science

    researchmap

  • Pinpoint drug delivery system using hollow bio-nanoparticles

    Tadanori Yamada, Masaharu Seno, Akihiko Kondo, Masakazu Ueda, Katsuyuki Tanizawa, Shun'ichi Kuroda

    Kobunshi Ronbunshu   61 ( 12 )   606 - 612   2004

     More details

    Language:Japanese   Publisher:Society of Polymer Science  

    Hepatitis B virus envelope L proteins produced in yeast cells form hollow nanoparticles (L particles, average diameter 220 nm) displaying human liver-specific receptor. Recently, the L particles were found to incorporate genes, proteins, and drugs, and act as an efficient pinpoint delivery system to human liver-derived tissues in xenograft models. By substituting the epidermal growth factor (EGF) for human liver-specific receptor, the mutated L particles showed the affinity to the EGF receptor, not to human liver. Other similar HBV envelope proteins, e.g., M and S particles, have already been commercialized for hepatitis B vaccine, strongly suggesting the safety of L particles in human. These results indicate that the hollow bio-nanoparticles are a promising candidate for the next-generation platform of DDS, especially that related to gene therapy.

    DOI: 10.1295/koron.61.606

    Scopus

    researchmap

  • Novel tissue and cell type-specific gene/drug delivery system using surface engineered hepatitis B virus nano-particles

    Tadanori Yamada, Masakazu Ueda, Masaharu Seno, Akihiko Kondo, Katsuyuki Tanizawa, Shun'ichi Kuroda

    Current Drug Targets - Infectious Disorders   4 ( 2 )   163 - 167   2004

     More details

    Language:English   Publishing type:Book review, literature introduction, etc.  

    The hepatitis B virus (HBV) surface antigen (HBsAg) L particle is a hollow nano-scale particle. HBsAg L particles have many properties that make them useful for in vivo gene transfer vectors and drug delivery systems. Gene therapy so far has required the in vivo pinpoint delivery of genetic materials into the target organs and cells. Gene transfer by HBsAg L particles might be an attractive method, since their tropism is the same as that of HBV. The HBsAg L particles are able to deliver therapeutic payloads with high specificity to human hepatocytes. In addition, the specificity of L particle can be altered by displaying various cell-binding molecules on the surface. Our results indicate that the L particle is suitable for a cell- and tissue-specific gene/drug transfer vector. In this review, we discuss HBsAg L particles as a gene/drug transfer vector and its potential for the treatment of infectious diseases. © 2004 Bentham Science Publishers Ltd.

    DOI: 10.2174/1568005043341037

    Scopus

    PubMed

    researchmap

  • 中空バイオナノ粒子が拓く新しい医療技術

    黒田俊一, 山田忠範, 妹尾昌治, 近藤昭彦, 上田政和, 谷澤克行

    化学工業   vol.55, no.12, pp.936-942   2004

     More details

  • Betacellulin improves glucose metabolism by promoting conversion of intraislet precursor cells to beta-cells in streptozotocin-treated mice

    L Li, M Seno, H Yamada, Kojima, I

    AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM   285 ( 3 )   E577 - E583   2003.9

     More details

    Language:English   Publisher:AMER PHYSIOLOGICAL SOC  

    Beta-cellulin (BTC) induces differentiation of pancreatic beta-cells and promotes regeneration of beta-cells in experimental diabetes. The present study was conducted to determine if BTC improved glucose metabolism in severe diabetes induced by a high dose of streptozotocin (STZ) in mice. Male ICR mice were injected with 200 mug/g ip STZ, and various doses of BTC were administered daily for 14 days. The plasma glucose concentration increased to a level of &gt;500 mg/dl in STZ-injected mice. BTC (0.2 mug/g) significantly reduced the plasma glucose concentration, but a higher concentration was ineffective. The effect of BTC was marked by day 4 but became smaller on day 6 or later. The plasma insulin concentration and the insulin content were significantly higher in mice treated with 0.1 and 0.2 mug/g BTC. BTC treatment significantly increased the number of beta-cells in each islet as well as the number of insulin-positive islets. Within islets, the numbers of 5-bromo-2-deoxyuridine/somatostatin-positive cells and pancreatic duodenal homeobox-1/somatostatin-positive cells were significantly increased by BTC. These results indicate that BTC improved hyperglycemia induced by a high dose of STZ by promoting neoformation of beta-cells, mainly from somatostatin-positive islet cells.

    DOI: 10.1152/ajpendo.00120.2003

    Web of Science

    researchmap

  • Nanoparticles for the delivery of genes and drugs to human hepatocytes

    T Yamada, Y Iwasaki, H Tada, H Iwabuki, MKL Chuah, T VandenDriessche, H Fukuda, A Kondo, M Ueda, M Seno, K Tanizawa, S Kuroda

    NATURE BIOTECHNOLOGY   21 ( 8 )   885 - 890   2003.8

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    Hepatitis B virus envelope L particles form hollow nanoparticles displaying a peptide that is indispensable for liver-specific infection by hepatitis B virus in humans. Here we demonstrate the use of L particles for the efficient and specific transfer of a gene or drug into human hepatocytes both in culture and in a mouse xenograft model. In this model, intravenous injection of L particles carrying the gene for green fluorescent protein (GFP) or a fluorescent dye resulted in observable fluorescence only in human hepatocellular carcinomas but not in other human carcinomas or in mouse tissues. When the gene encoding human clotting factor IX was transferred into the xenograft model using L particles, factor IX was produced at levels relevant to the treatment of hemophilia B. The yeast-derived L particle is free of viral genomes, highly specific to human liver cells and able to accommodate drugs as well as genes. These advantages should facilitate targeted delivery of genes and drugs to the human liver.

    DOI: 10.1038/nbt843

    Web of Science

    researchmap

  • バイオナノ 粒子を用いる遺伝子・薬剤のピンポイントドラッグデリバリーシステム

    妹尾昌治, 黒田俊一, 近藤昭彦, 多田宏子, 谷澤克行, 上田政和

    バイ オインダストリー   20(4):54-64   2003

     More details

  • 「中空バイオナノ粒子に よるピンポイントドラッグデリバリーシステム」

    近藤昭彦, 黒田俊一, 谷澤克行, 妹尾昌治, 上田政和

    化学工学   67巻12号 649-714頁   2003

     More details

  • The cytotoxicity of a conjugate composed of human epidermal growth factor and eosinophil cationic protein

    H Jinno, M Ueda, S Ozawa, T Ikeda, M Kitajima, T Maeda, M Seno

    ANTICANCER RESEARCH   22 ( 6C )   4141 - 4145   2002.11

     More details

    Language:English   Publisher:INT INST ANTICANCER RESEARCH  

    Background: Conventional targeted therapy with foreign proteins is highly immunogenic. In this study, we developed targeted therapy composed of human endogenous proteins and evaluated its efficacy in vitro. Materials and Methods: Human epidermal growth factor (EGF) was chemically linked to human eosinophil cationic protein (ECP). The cytotoxicity of the EGF-ECP conjugate was evaluated by MTT assay. Results: The conjugate showed dose-dependent cytotoxicity on EGF receptor (EGFR)-overexpressing BT-20 cells with an IC50 of 1.5 x 10(-7) M, whereas the IC50 of ECP alone was almost 10(-4) M, The conjugate had no detectable cytotoxicity against EGF receptor- deficient H69 cells. Excess EGF protected BT-20 cells from the cytotoxicity of the conjugate. Comparing the cytotoxicity and the level of EGFR expression, the cytotoxicity of the conjugate was positively correlated with the level of EGFR expression of each cell line. Conclusion: Conjugates composed solely of human proteins might be useful with less immunogenicity and less toxicity than the conventional immunotoxins for targeted therapy.

    Web of Science

    researchmap

  • RNase 3 (ECP) is an extraordinarily stable protein among human pancreatic-type RNases

    T Maeda, K Mahara, M Kitazoe, J Futami, A Takidani, M Kosaka, H Tada, M Seno, H Yamada

    JOURNAL OF BIOCHEMISTRY   132 ( 5 )   737 - 742   2002.11

     More details

    Language:English   Publisher:JAPANESE BIOCHEMICAL SOC  

    There have been some attempts to develop immunotoxins utilizing human RNase as a cytotoxic domain of antitumor agents. We have recently shown that only human RNase 3 (eosinophil cationic protein, ECP) among five human pancreatic-type RNases excels in binding to the cell surface and has a growth inhibition effect on several cancer cell lines, even though the RNase activity of RNase 3 is completely inhibited by the ubiquitously expressed cytosolic RNase inhibitor. This phenomenon may be explained by that RNase 3 is very stable against proteolytic degradation because RNase 3 internalized through endocytosis could have a longer life time in the cytosol, resulting in the accumulation of enough of it to exceed the concentration of RNase inhibitor, which allows the degradation of cytosolic RNA molecules. Thus, we compared the stabilities of human pancreatic-type RNases (RNases 1-5) and bovine RNase A by means of guanidium chloride-induced denaturation experiments based on the assumption of a two-state transition for unfolding. It was demonstrated that RNase 3 is extraordinarily stabler than either RNase A or the other human RNases (by more than 25 kJ/mol). Thus, our data suggest that in addition to its specific affinity for certain cancer cell lines, the stability of RNase 3 contributes to its unique cytotoxic effect and that it is important to stabilize a human RNase moiety through protein engineering for the design of human RNase-based immunotoxins.

    Web of Science

    researchmap

  • Optimum modification for the highest cytotoxicity of cationized ribonuclease

    J Futami, E Nukui, T Maeda, M Kosaka, H Tada, M Seno, H Yamada

    JOURNAL OF BIOCHEMISTRY   132 ( 2 )   223 - 228   2002.8

     More details

    Language:English   Publisher:JAPANESE BIOCHEMICAL SOC  

    Cationization of a protein is considered to be a powerful strategy for internalizing a functional protein into cells. Cationized proteins appear to adsorb to the cell surface by electrostatic interactions, then enter the cell in a receptor- and transporter-independent fashion. Thus, in principle, all cell types appear to take up cationized proteins. Since ribonucleases (RNases) have a latent cytotoxic potential, cationized RNases could be useful cancer chemotherapeutics. In this study, we investigated the effect of the degree of cationization on the cytotoxicity of RNase A by modifying carboxyl groups with ethylenediamine. We found that there is an optimum degree of modification for cytotoxicity, in which 5 to 7 out of 11 carboxyl groups in RNase A are modified, toward MCF-7 and 3T3-SV-40 cells. More interestingly, the cytotoxicity of cationized RNase As correlates well with the value of [RNase activity] x [estimated concentration of RNase free from RNase inhibitor], mimicking the practical enzymatic activity of cationized RNase As in cytosol. The results indicate that cationization of a protein to an optimum level is important for maintaining protein function in the cytosol. Sophisticated protein cationization techniques will help to advance protein transduction technology.

    Web of Science

    researchmap

  • Solution structure of betacellulin, a new member of EGF-family ligands

    K Miura, H Doura, T Aizawa, H Tada, M Seno, H Yamada, K Kawano

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   294 ( 5 )   1040 - 1046   2002.6

     More details

    Language:English   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The solution structure of the EGF-like domain of betacellulin (BTCe), a newly discovered member of the epidermal growth factor (EGF) family, has been determined using two-dimensional nuclear magnetic resonance spectroscopy. This is the first report to identify the solution structure of the EGF-family ligand monomers that interact with both ErbB-1 and ErbB-4. The solution structure of BTCe was calculated using 538 NMR-derived restraints. The overall structure of BTCe was stabilized by three disulfide bonds, a hydrophobic core, and 23 hydrogen bonds. It appears that BTCe is comprised of five beta-strands and one short 3(10) helical turn. The secondary structural elements of BTCe are basically similar to those of the other EGF-family proteins, except that several significant variations of the structural properties were found. It is suggested that the structural variations between BTCe and the other EGF-family ligands may affect the specific receptor-recognition properties of EGF-family ligands. (C) 2002 Elsevier Science (USA). All rights reserved.

    DOI: 10.1016/S0006-291X(02)00585-5

    Web of Science

    researchmap

  • Combined expression of pancreatic duodenal homeobox 1 and islet factor 1 induces immature enterocytes to produce insulin

    H Kojima, T Nakamura, Y Fujita, A Kishi, M Fujimiya, S Yamada, M Kudo, Y Nishio, H Maegawa, M Haneda, H Yasuda, Kojima, I, M Seno, NCW Wong, R Kikkawa, A Kashiwagi

    DIABETES   51 ( 5 )   1398 - 1408   2002.5

     More details

    Language:English   Publisher:AMER DIABETES ASSOC  

    Immature rat intestinal stem cells (IEC-6) given the ability to express the transcription factor, pancreatic duodenal homeobox 1 (Pdx-1), yielded YK cells. Although these cells produced multiple enteroendocrine hormones, they did not produce insulin. Exposure of YK cells to 2 nmol/l betacellulin yielded BYK cells that showed the presence of insulin expression in cytoplasm and that secreted insulin into culture media. By examining the mechanism of differentiation in BYK cells, we found that another transcription factor, islet factor 1 (Isl-1) was newly expressed with the disappearance of Pax-6 expression in those cells after exposure to betacellulin. These results indicated that combined expression of Pdx-1 and Isl-1 in IEC-6 cells was required for the production of insulin. In fact, overexpression of both Pdx-1 and Isl-1 in IEC-6 cells (Isl-YK-12, -14, and -15 cells) gave them the ability to express insulin without exposure to betacellulin. Furthermore, implantation of the Isl-YK-14 cells into diabetic rats reduced the animals' plasma glucose levels; glucose levels dropped from 19.4 to 16.9 mmol/l 1 day after the injection of cells. As expected, the plasma insulin concentrations were 2.7 times higher in the diabetic rats injected with Isl-YK-14 cells compared to in controls. In summary, our results indicated that immature intestinal stem cells can differentiate into insulin-producing cells given the ability to express the transcription factors Pdx-1 and Isl-1.

    Web of Science

    researchmap

  • Cripto-1 activates nodal- and ALK4-dependent and -independent signaling pathways in mammary epithelial cells

    C Bianco, HB Adkins, C Wechselberger, M Seno, N Normanno, A De Luca, YP Sun, N Khan, N Kenney, A Ebert, KP Williams, M Sanicola, DS Salomon

    MOLECULAR AND CELLULAR BIOLOGY   22 ( 8 )   2586 - 2597   2002.4

     More details

    Language:English   Publisher:AMER SOC MICROBIOLOGY  

    Cripto-1 (CR-1), an epidermal growth factor-CFC (EGF-CFC) family member, has a demonstrated role in embryogenesis and mammary gland development and is overexpressed in several human tumors. Recently, EGF-CFC proteins were implicated as essential signaling cofactors for Nodal, a transforming growth factor beta family member whose expression has previously been defined as embryo specific. To identify a receptor for CR-1, a human brain cDNA phage display library was screened using CR-1 protein as bait. Phage inserts with identity to ALK4, a type I serine/threonine kinase receptor for Activin, were identified. CR-1 binds to cell surface ALK4 expressed on mammalian epithelial cells in fluorescence-activated cell sorter analysis, as well as by coimmunoprecipitation. Nodal is coexpressed with mouse Cr-1 in the mammary gland, and CR-1 can phosphorylate the transcription factor Smad-2 in EpH-4 mammary epithelial cells only in the presence of Nodal and ALK4. In contrast, CR-1 stimulation of mitogen-activated protein kinase and AKT in these cells is independent of Nodal and ALK4, suggesting that CR-1 may modulate different signaling pathways to mediate its different functional roles.

    DOI: 10.1128/MCB.22.8.2586-2597.2002

    Web of Science

    researchmap

  • Growth inhibition of mammalian cells by eosinophil cationic protein

    T Maeda, M Kitazoe, H Tada, R de Llorens, DS Salomon, M Ueda, H Yamada, M Seno

    EUROPEAN JOURNAL OF BIOCHEMISTRY   269 ( 1 )   307 - 316   2002.1

     More details

    Language:English   Publisher:BLACKWELL PUBLISHING LTD  

    Eosinophil cationic protein (ECP), one of the major components of basic granules of eosinophils, is cytotoxic to tracheal epithelium. However, the extent of this effect on other cell types has not been evaluated in vitro. In this study, we evaluated the effect of ECP on 13 mammalian cell lines. ECP inhibited the growth of several cell lines including those derived from carcinoma and leukemia in a dose-dependent manner. The IC50 values on A431 cells, MDA-MB-453 cells, HL-60 cells and K562 cells were estimated to be approximate to1-5 muM. ECP significantly suppressed the size of colonies of A431 cells, and decreased K562 cells in G(1)/G(0) phase. However, there was little evidence that ECP killed cells in either cell line. These effects of ECP were not enhanced by extending its N-terminus. Rhodamine B isothiocyanate-labeled ECP started to bind to A431 cells after 0.5 h and accumulated for up to 24 h, indicating that specific affinity for the cell surface may be important. ne affinity of ECP for heparin was assessed and found to be reduced when tryptophan residues, one of which is located at a position in the catalytic subsite of ribonuclease in ECP, were modified. The growth-inhibitory effect was also attenuated by this modification. These results suggest that growth inhibition by ECP is dependent on cell type and is cytostatic.

    DOI: 10.1046/j.0014-2956.2001.02653.x

    Web of Science

    researchmap

  • Detection and localization of Cripto-1 binding in mouse mammary epithelial cells and in the mouse mammary gland using an immunoglobulin-Cripto-1 fusion protein

    C Bianco, N Normanno, A De Luca, MR Maiello, C Wechselberger, Y Sun, N Khan, H Adkins, M Sanicola, B Vonderhaar, B Cohen, M Seno, D Salomon

    JOURNAL OF CELLULAR PHYSIOLOGY   190 ( 1 )   74 - 82   2002.1

     More details

    Language:English   Publisher:WILEY-LISS  

    Human Cripto-1 (CR-1), a member of the epidermal growth factor-CFC (EGF-CFC) family of peptides, is expressed in the developing mouse mammary gland and can modulate mammary epithelial cell migration, branching morphogenesis and milk protein expression in vitro. In order to screen for a CR-1 receptor and to identify potential CR-1 target tissues, we constructed a fusion protein comprising the EGF-like domain of CR-1 and the Fc domain of a human IgG1. The recombinant CR-1 fusion protein (CR-1-Fc) was biologically active as it was able to activate the ras/raf/mitogen activated protein kinase (MAPK) pathway and to inhibit transcription of the milk protein P-casein in NMuMG and HC-11 mouse mammary epithelial cells. By using immunocytochemistry and by an in situ enzyme-linked immunosorbent assay (ELISA), CR-1-Fc was found to specifically bind to NMuMG and HC-11 cells. Finally, immunohistochemical analysis using CR-1-Fc showed a specific localization of CR-1 binding to tissue sections from mouse mammary gland. In particular, more than 60% of the epithelial cells were intensely stained with the CR-1-Fc fusion protein in the lactating mouse mammary gland, whereas approximately 25% of the mammary epithelial cells were stained in the gland from pregnant mouse. Since expression of mouse cripto-1 (Cr-1) in the pregnant and lactating mouse mammary gland as well as its presence in milk has been previously demonstrated, these data strongly suggest that an autocrine pathway involving Cr-1 and its putative receptor is operating in the mouse mammary gland during pregnancy and lactation. Published 2002 Wiley-Liss, Inc.(dagger)

    DOI: 10.1002/jcp.10037

    Web of Science

    researchmap

  • 細胞増殖因子ベータセルリン -膵臓再生と糖尿病研究へのニューマテリアル-

    妹尾昌治

    和光純薬時報   70(3): 6-8.   2002

     More details

  • Human betacellulin structure modeled from other members of EGF family

    Lopez-Torrejon, I, E Querol, FX Aviles, M Seno, R de Llorens, B Oliva

    JOURNAL OF MOLECULAR MODELING   8 ( 4 )   131 - 144   2002

     More details

    Language:English   Publisher:SPRINGER-VERLAG  

    We have modeled betacellulin (BTC) to gain insight into the structural elements that can explain its properties. The epidermal growth factor (EGF) signal transduction pathway, a significant mediator of several cell functions, is based on four closely related tyrosine kinase receptors. The ErbB receptors are transmembrane glycoproteins and signal transduction is initiated by ligand binding that induces receptor homo- or heterodimerization to form a complex containing two molecules of ligand and two molecules of receptor. The EGF family of ligands can be divided into three groups based on their ability to bind and activate distinct ErbB receptor homo- and heterodimers. Each member of the group formed by BTC, heparin binding EGF (HB-EGF) and epiregulin (EP) can interact with both the EGF receptor (EGFR) and heregulin receptors (ErbB-3 and ErbB-4), and are hence called "bispecific" ligands. BTC and EP also present the distinctive feature that they activate all possible heterodimeric ErbB receptors. BTC has been modeled with the program MODELLER, using human EGF, human transforming growth factor alpha (hTGFalpha), human HB-EGF and human heregulin one alpha (hHRG-1alpha) as templates. The structure of the model as well as that of the templates were optimized and a simulation of 100 ps was run for all. The main structural properties of the model and the templates were compared and in conclusion the hBTC conformation was closely similar to that of hTGFalpha. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00894-002-0072-2.

    DOI: 10.1007/s00894-002-0072-2

    Web of Science

    researchmap

  • Promotion of beta-cell regeneration by betacellulin in ninety percent-pancreatectomized rats

    L Li, M Seno, H Yamada, Kojima, I

    ENDOCRINOLOGY   142 ( 12 )   5379 - 5385   2001.12

     More details

    Language:English   Publisher:ENDOCRINE SOC  

    Betacellulin is thought to promote growth and differentiation of pancreatic beta -cells. We investigated the effect of betacellulin on regeneration of pancreatic beta -cells in 90%-pancreatectomized rats. Ninety percent pancreatectomy was performed in male Wistar rats and betacellulin (0.5 mug/g body weight) or saline was administered daily for 10 d starting immediately after pancreatectomy. In pancreatectomized rats, the morning-fed plasma glucose was significantly lower and the plasma insulin concentration was significantly higher in betacellulin-treated rats than those in control rats for up to 4 wk. Thirty days after pancreatectomy, a glucose tolerance test was performed. Betacellulin reduced the plasma glucose response to ip glucose loading. In control rats, the plasma insulin concentration was significantly lower and did not respond to glucose. In contrast, the plasma insulin concentration increased slightly but significantly in betacellulin-treated rats. Thirty days after pancreatectomy, the beta -cell mass was greater and the insulin content was significantly higher in betacellulin-treated rats than those in control rats. The numbers of islet cell-like cluster and bromodeoxy uridine/insulin double-positive cells in both islet cell-like cluster and islets were significantly higher in betacellulin-treated rats. These results indicate that administration of betacellulin improves glucose metabolism by promoting beta -cell regeneration in 90%-pancreatectomized rats.

    DOI: 10.1210/en.142.12.5379

    Web of Science

    researchmap

  • Preparation of potent cytotoxic ribonucleases by cationization: Enhanced cellular uptake and decreased interaction with ribonuclease inhibitor by chemical modification of carboxyl groups

    J Futami, T Maeda, M Kitazoe, E Nukui, H Tada, M Seno, M Kosaka, H Yamada

    BIOCHEMISTRY   40 ( 25 )   7518 - 7524   2001.6

     More details

    Language:English   Publisher:AMER CHEMICAL SOC  

    Carboxyl groups of bovine RNase A were amidated with ethylenediamine (to convert negative charges of carboxylate anions to positive ones), 2-aminoethanol (to eliminate negative charges), and taurine (to keep negative charges), respectively, by a carbodiimide reaction. Human RNase 1 was also modified with ethylenediamine. Surprisingly, the modified RNases were all cytotoxic toward 3T3-SV-40 cells despite their decreased ribonucleolytic activity. However, their enzymatic activity was not completely eliminated by the presence of excess cytosolic RNase inhibitor (RI). As for native RNase A and RNase 1 which were not cytotoxic, they were completely inactivated by RI. More interestingly, within the cytotoxic RNase derivatives. cytotoxicity correlated well with the net positive charge. RNase 1 and RNase A modified with ethylenediamine were more cytotoxic than naturally occurring cytotoxic bovine seminal RNase. An experiment using the fluorescence-labeled RNase derivatives indicated that the more cationic RNases were more efficiently adsorbed to the cells. Thus, it is suggested that the modification of carboxyl groups could change complementarity of RNase to RI and as a result endow RNase cytotoxicity and that cationization enhances the efficiency of cellular uptake of RNase so as to strengthen its cytotoxicity. The finding that an extracellular human enzyme such as RNase 1 could be effectively internalized into the cell by cationization suggests that cationization is a simple strategy for efficient delivery of a protein into cells and may open the way of the development of new therapeutics.

    DOI: 10.1021/bi010248g

    Web of Science

    researchmap

  • Three-dimensional structure of human RNase 1 Delta N7 at 1.9 angstrom resolution

    J Pous, G Mallorqui-Fernandez, R Peracaula, SS Terzyan, J Futami, H Tada, H Yamada, M Seno, R de Llorens, FX Gomis-Ruth, M Coll

    ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY   57   498 - 505   2001.4

     More details

    Language:English   Publisher:MUNKSGAARD INT PUBL LTD  

    Human pancreatic ribonuclease 1 (RNase 1) is considered to be the human counterpart of bovine pancreatic RNase A. Truncation of seven amino-acid residues from the aminoterminal sequence resulted in RNase 1 Delta N7, which has a reduced ribonucleolytic activity and a lower affinity for the human placental RNase inhibitor (PRI). This RNase 1 variant has been cloned, heterologously overexpressed, purified and crystallized. Its crystal structure has been determined and refined using data to 1.9 Angstrom resolution. The molecule displays the alpha + beta folding topology typical of members of the RNase A superfamily. The main distinct features found in RNase 1 Delta N7 are basically located in three loops affecting the fitting of the enzyme to the active site of subtilisin and the shape of the B2 subsite. These changes, taken with the lack of the catalytically active residue Lys7, may explain the reduced affinity of RNase 1 Delta N7 for PRI and the low ribonucleolytic activity of the protein when compared with the native enzyme.

    DOI: 10.1107/S0907444901001147

    Web of Science

    researchmap

  • Physicochemical and immunological characterization of hepatitis B virus envelope particles exclusively consisting of the entire L (pre-S1+pre-S2+S) protein

    T Yamada, H Iwabuki, T Kanno, H Tanaka, T Kawai, H Fukuda, A Kondo, M Seno, K Tanizawa, S Kuroda

    VACCINE   19 ( 23-24 )   3154 - 3163   2001.4

     More details

    Language:English   Publisher:ELSEVIER SCI LTD  

    The hepatitis B virus (HBV) envelope (env) protein is composed of three regions: the 108- or 119-residue pre-Si region involved in the direct interaction with hepatocytes. the 55-residue pre-SZ region associated with the polymerized albumin-mediated interaction, and the major 226-residue S protein region. Thus, to improve the immunogenic potency of conventional HE vaccines, development of a new vaccine containing the entire pre-Si region in addition to pre-S2 and S is desired. We previously reported the efficient production of the HBV env L (pre-S1 + pre-S2 + S) protein in the recombinant yeast cells [J Biol Chem 267 (1992) 1953]. In this study, the HBV env L protein produced as nano-particles in yeast has been purified and characterized. By equilibrium sedimentation. an average molecular weight of L particle was estimated to be approximately 6.3 x 10(6), indicating that about 110 molecules of L proteins are assembled into an L particle. By atomic force microscopy in a moist atmosphere. the L particles were observed as large spherical particles with a diameter of 50-500 nm. The L particles were stable on short-time heating at a high temperature and long-time storage at a low temperature but rather unstable on repeated freezing and thawing and treatment with dithiothreitol. When immunized in mice. L particles elicited efficiently and simultaneously the anti-g, anti-pre-S2, and anti-pre-Si antibodies. The ED50 values in mice for the anti-S and anti-pre-S2 antibodies were similar to those elicited by the M (pre-S2 + S) particles. Furthermore. the anti-pre-Si rabbit antibodies were found to recognize various segments of the pre-Si region, including the pre-Si (21-47) segment. These results show the high ability of L particles to induce all antibodies against HBV env proteins, hence promising the future application of L particles for the next generation HE vaccine. (C) 2001 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0264-410X(01)00017-2

    Web of Science

    researchmap

  • ベータセルリン:細胞増殖分化因子を応用した再生医療への試み

    妹尾昌治

    ハイテクインフォメーション   134号14-21   2001

     More details

  • Expression of phospholipid hydroperoxide glutathione peroxidase (PHGPx) mRNA in rat testes

    H Mori, T Nomura, M Seno, Y Miki, T Kimura, T Kogami, J Sasaki

    ACTA HISTOCHEMICA ET CYTOCHEMICA   34 ( 1 )   25 - 30   2001

     More details

    Language:English   Publisher:JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY  

    We investigated the expression of reactive oxygen species (ROS)-related enzyme mRNA to clarify the role of ROS in reproduction. In the present study, we investigated the expression of phospholipid hydroperoxide glutathione peroxidase (PHGPx) mRNA in rat testes by in situ hybridization. To prepare digoxigenin-labeled probes, we obtained 5 ' -phosphorylated oligonucleotides containing sense and antisense sequences (93-mer) with EcoR I or Hind III restriction sites as protruding cohesive ends (total 99-mer). Then, both oligonucleotides were annealed and cloned into a pGEM4Z vector. The resultant plasmid DNA was linearized with EcoR I or Hind III and used as a template for the synthesis of labeled sense or antisense riboprobes.
    PHGPx mRNA was expressed stage-specifically during spermatogenesis in adult rats. It was not expressed during spermatogonia, but its expression first appeared in stage VII pachytene spermatocytes, became evident in stage VIII pachytene spermatocytes and increased gradually until becoming diplotene spermatocytes. It decreased slightly during the metaphase of meiosis, and then gradually increased as the spermatids differentiated. The expression was marked in spermatids between steps 7 and 12, and the maximum expression was observed in step 9-10 spermatids. Expression was diminished completely in step 18 spermatids. These results suggest that PHGPx plays an essential role in spermatogenesis.

    Web of Science

    researchmap

  • The EGF-CFC family: novel epidermal growth factor-related proteins in development and cancer

    DS Salomon, C Bianco, AD Ebert, NI Khan, M De Santis, N Normanno, C Wechselberger, M Seno, K Williams, M Sanicola, S Foley, W Gullick, G Persico

    ENDOCRINE-RELATED CANCER   7 ( 4 )   199 - 226   2000.12

     More details

    Language:English   Publishing type:Book review, literature introduction, etc.   Publisher:SOC ENDOCRINOLOGY  

    The EGF-CFC gene family encodes a group of structurally related proteins that serve as important competence factors during early embryogenesis in Xenopus, zebrafish, mice and humans. This multigene family consists of Xenopus FRL-1, zebrafish one-eyed-pinhead (oep), mouse cripto (Cr-l) and cryptic, and human cripto (CR-1) and criptin. FRL-1, oep and mouse cripto are essential for the formation of mesoderm and endoderm and for correct establishment of the anterior/posterior axis. In addition, oep and cryptic are important for the establishment of left-right (UR) asymmetry, In zebrafish, there is strong genetic evidence that oep functions as an obligatory cc-factor for the correct signaling of a transforming growth factor-beta (TGF beta)-related gene, nodal, during gastrulation and during UR asymmetry development. Expression of Cr-l and cryptic is extinguished in the embryo after day 8 of gestation except for the developing heart where Cr-l expression is necessary for myocardial development. In the mouse, cryptic is not expressed in adult tissues whereas Cr-l is expressed at a low level in several different tissues including the mammary gland, In the mammary gland, expression of Cr-l in the ductal epithelial cells increases during pregnancy and lactation and immunoreactive and biologically active Cr-l protein can be detected in human milk. Overexpression of Cr-l in mouse mammary epithelial cells can facilitate their in vitro transformation and in vivo these Cr-l-transduced cells produce ductal hyperplasias in the mammary gland. Recombinant mouse or human cripto can enhance cell motility and branching morphogenesis in mammary epithelial cells and in some human tumor cells. These effects are accompanied by an epithelial-mesenchymal transition which is associated with a decrease in beta -catenin function and an increase in vimentin expression. Expression of cripto is increased several-fold in human colon, gastric, pancreatic and lung carcinomas and in a variety of different types of mouse and human breast carcinomas. More importantly, this increase can first be detected in premalignant lesions in some of these tissues. Although a specific receptor for the EGF-CFC proteins has not yet been identified, oep depends upon an activin-type RIIB and RIB receptor system that functions through Smad-e. Mouse and human cripto have been shown to activate a ras/raf/MAP kinase signaling pathway in mammary epithelial cells. Activation of phosphatidylinositol 3-kinase and Akt are also important for the ability of CR-1 to stimulate cell migration and to block lactogenic hormone-induced expression of p-casein and whey acidic protein. In mammary epithelial cells, part of these responses may depend on the ability of CR-1 to transactivate erb B-4 and/or fibroblast growth factor receptor 1 through an src-like tyrosine kinase.

    DOI: 10.1677/erc.0.0070199

    Web of Science

    researchmap

  • Stabilization of human RNase 1 by introduction of a disulfide bond between residues 4 and 118

    J Futami, H Tada, M Seno, S Ishikami, H Yamada

    JOURNAL OF BIOCHEMISTRY   128 ( 2 )   245 - 250   2000.8

     More details

    Language:English   Publisher:JAPANESE BIOCHEMICAL SOC  

    In order to stabilize human RNase 1 by introduction of an intramolecular cross-link, a mutant protein (4-118CL RNase 1), in which Arg4 and Val118 are replaced with cysteine residues and linked by a disulfide bond, was designed and expressed in Escherichia coli as inclusion bodies. The 4-118CL RNase 1 that refolded under redox conditions was a monomer without free SH groups and retained 11% of the activity of the wild-type recombinant RNase 1, indicating that the mutant enzyme was correctly folded with the formation of an additional disulfide bond between Cys4 and Cys118, From guanidium chloride denaturation experiments based on the assumption of a two-state transition for unfolding, it was demonstrated that the introduction of the present cross-link increased the thermodynamic stability of RNase 1 by 2.0 kcal/mol. This value was lower than that, 5.4 kcal/mol, theoretically calculated from the reduction of chain entropy of the unfolded state due to the introduction of the cross-link. These results suggest that the present cross-link also destabilized the folded state of RNase 1 by 3.4 kcal/mol. Along with the increase in the thermodynamic stability, the stability of RNase 1 against trypsin digestion was also significantly increased by the introduction of this cross-link. It is likely, although not proven, that stabilized human RNases are favorable for clinical use, because human RNase-based immunotoxins should have long half-lives as to proteolytic degradation after endocytosis.

    Web of Science

    researchmap

  • Three-dimensional crystal structure of human eosinophil cationic protein (RNase 3) at 1.75 Å resolution

    Goretti Mallorquí-Fernández, Joan Pous, Rosa Peracaula, Joan Aymamí, Takashi Maeda, Hiroko Tada, Hidenori Yamada, Masaharu Seno, Rafael De Llorens, F.Xavier Gomis-Rüth, Miquel Coll

    Journal of Molecular Biology   300 ( 5 )   1297 - 1307   2000.7

     More details

    Language:English   Publisher:Academic Press  

    Eosinophil cationic protein (ECP
    RNase 3) is a human ribonuclease found only in eosinophil leukocytes that belongs to the RNase A superfamily. This enzyme is bactericidal, helminthotoxic and cytotoxic to mammalian cells and tissues. The protein has been cloned, heterologously overexpressed, purified and crystallized. Its crystal structure has been determined and refined using data up to 1.75 Å resolution. The molecule displays the α + β folding topology typical for members of the ribonuclease A superfamily. The catalytic active site residues are conserved with respect to other ribonucleases of the superfamily but some differences appear at substrate recognition subsites, which may account, in part, for the low catalytic activity. Most strikingly, 19 surface-located arginine residues confer a strong basic character to the protein. The high concentration of positive charges and the particular orientation of the side-chains of these residues may also be related to the low activity of ECP as a ribonuclease and provides an explanation for its unique cytotoxic role through cell membrane disruption. (C) 2000 Academic Press.

    DOI: 10.1006/jmbi.2000.3939

    Scopus

    PubMed

    researchmap

  • Three-dimensional crystal structure of human eosinophil cationic protein (RNase 3) at 1.75 Å resolution

    Goretti Mallorquí-Fernández, Joan Pous, Rosa Peracaula, Joan Aymamí, Takashi Maeda, Hiroko Tada, Hidenori Yamada, Masaharu Seno, Rafael De Llorens, F.Xavier Gomis-Rüth, Miquel Coll

    Journal of Molecular Biology   300 ( 5 )   1297 - 1307   2000.7

     More details

    Language:English   Publisher:Academic Press  

    Eosinophil cationic protein (ECP
    RNase 3) is a human ribonuclease found only in eosinophil leukocytes that belongs to the RNase A superfamily. This enzyme is bactericidal, helminthotoxic and cytotoxic to mammalian cells and tissues. The protein has been cloned, heterologously overexpressed, purified and crystallized. Its crystal structure has been determined and refined using data up to 1.75 Å resolution. The molecule displays the α + β folding topology typical for members of the ribonuclease A superfamily. The catalytic active site residues are conserved with respect to other ribonucleases of the superfamily but some differences appear at substrate recognition subsites, which may account, in part, for the low catalytic activity. Most strikingly, 19 surface-located arginine residues confer a strong basic character to the protein. The high concentration of positive charges and the particular orientation of the side-chains of these residues may also be related to the low activity of ECP as a ribonuclease and provides an explanation for its unique cytotoxic role through cell membrane disruption. (C) 2000 Academic Press.

    DOI: 10.1006/jmbi.2000.3939

    Scopus

    PubMed

    researchmap

  • Three-dimensional crystal structure of human eosinophil cationic protein (RNase 3) at 1.75 angstrom resolution

    G Mallorqui-Fernandez, J Pous, R Peracaula, J Aymami, T Maeda, H Tada, H Yamada, M Seno, R de Llorens, FX Gomis-Ruth, M Coll

    JOURNAL OF MOLECULAR BIOLOGY   300 ( 5 )   1297 - 1307   2000.7

     More details

    Language:English   Publisher:ACADEMIC PRESS LTD  

    Eosinophil cationic protein (ECP; RNase 3) is a human ribonuclease found only in eosinophil leukocytes that belongs to the RNase A superfamily. This enzyme is bactericidal, helminthotoxic and cytotoxic to mammalian cells and tissues. The protein has been cloned, heterologously overexpressed, purified and crystallized. Its crystal structure has been determined and refined using data up to 1.75 Angstrom resolution. The molecule displays the alpha + beta folding topology typical for members of the ribonuclease A superfamily. The catalytic active site residues are conserved with respect to other ribonucleases of the superfamily but some differences appear at substrate recognition subsites, which may account, in part, for the low catalytic activity. Most strikingly, 19 surface-located arginine residues confer a strong basic character to the protein. The high concentration of positive charges and the particular orientation of the side-chains of these residues may also be related to the low activity of ECP as a ribonuclease and provides an explanation for its unique cytotoxic role through cell membrane disruption. (C) 2000 Academic Press.

    DOI: 10.1006/jmbi.2000.3939

    Web of Science

    researchmap

  • Molecular cloning and expression of rat betacellulin cDNA

    Hiroko Tada, Masaharu Seno, Hidenori Yamada, Reiko Sasada, Koichi Igarashi

    Biochimica et Biophysica Acta - Gene Structure and Expression   1492 ( 1 )   285 - 288   2000.6

     More details

    Language:English  

    The cDNA encoding an entire open reading frame of rat betacellulin has been cloned from rat kidney. Expression of this cDNA in COS7 cells showed a significant amount of mitogenic activity in the culture media. Western blotting of the cell lysates suggested that the membrane-anchored precursor was cleaved to release its ectodomain very efficiently. Copyright (C) 2000.

    DOI: 10.1016/S0167-4781(00)00106-8

    Scopus

    PubMed

    researchmap

  • Molecular cloning and expression of rat betacellulin cDNA

    Hiroko Tada, Masaharu Seno, Hidenori Yamada, Reiko Sasada, Koichi Igarashi

    Biochimica et Biophysica Acta - Gene Structure and Expression   1492 ( 1 )   285 - 288   2000.6

     More details

    Language:English  

    The cDNA encoding an entire open reading frame of rat betacellulin has been cloned from rat kidney. Expression of this cDNA in COS7 cells showed a significant amount of mitogenic activity in the culture media. Western blotting of the cell lysates suggested that the membrane-anchored precursor was cleaved to release its ectodomain very efficiently. Copyright (C) 2000.

    DOI: 10.1016/S0167-4781(00)00106-8

    Scopus

    PubMed

    researchmap

  • Molecular cloning and expression of rat betacellulin cDNA

    H Tada, M Seno, H Yamada, R Sasada, K Igarashi

    BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION   1492 ( 1 )   285 - 288   2000.6

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    The cDNA encoding an entire open reading frame of rat betacellulin has been cloned from rat kidney. Expression of this cDNA in COS7 cells showed a significant amount of mitogenic activity in the culture media. Western blotting of the cell lysates suggested that the membrane-anchored precursor was cleaved to release its ectodomain very efficiently. (C) 2000 Published by Elsevier Science B.V.

    DOI: 10.1016/S0167-4781(00)00106-8

    Web of Science

    researchmap

  • Targeting activated lymphocytes with an entirely human immunotoxin analogue: Human pancreatic RNase1-human IL-2 fusion

    K Psarras, M Ueda, M Tanabe, M Kitajima, S Aiso, S Komatsu, M Seno

    CYTOKINE   12 ( 6 )   786 - 790   2000.6

     More details

    Language:English   Publisher:W B SAUNDERS CO  

    A hybrid human protein was produced in E. coli by fusing the genes encoding human pancreatic RNase1 (hpRNase1) and human IL-2 (hIL-2), The recombinant hpRNase1-hIL-2 inhibited protein synthesis in HTLV-l-infected, malignant T cells, which hyperproduce high affinity IL-2 receptors, with an IC50 of 2 x 10 (- 8) M, whereas no inhibition was detectable in control cells with lower affinity receptors, HpRNase1 alone had an IC50 of almost 10 (- 3) M. A molar excess of hIL-2 blocked the protein synthesis inhibition dose-dependently. In a human mixed lymphocyte culture, hpRNase1-hIL-2 inhibited the proliferation of responder cells with potency comparable to that of cyclosporine, while non-effective doses of FK506 importantly improved its potency. Despite its short half-life in animals, hpRNase1-hIL-2 rapidly enters cells in a few minutes and arrests the protein translation in less than 10 h. Thus, hpRNase1-hIL-2 may be useful to selectively eliminate activated lymphocytes hyperproducing high affinity 1L-2 receptors, as in allograft rejection, graft-versus-host disease, autoimmune disorders, adult T cell leukaemia and other lymphoproliferative or retroviral malignancies including HIV infection, without inducing general immunosuppression, As an entirely human "immunotoxin analogue" it may alleviate the dose limiting toxicity and immunogenicity of conventional immunotoxins. (C) 2000 Academic Press.

    DOI: 10.1006/cyto.1999.0619

    Web of Science

    researchmap

  • Stabilization of human RNase 1 by introduction of a disulfide bond between residues 4 and 118.

    Futami J, Tada H, *Seno M, Ishikami S, Yamada H.

    J Biochem (Tokyo)   128 ( 2 )   245   2000.4

     More details

  • Convenient and Efficient In Vitro Folding of Disulfide-Containing Globular Protein from Crude Bacterial Inclusion Bodies.

    J Biochem (Tokyo)   127 ( 3 )   435   2000.4

     More details

  • Convenient and Efficient In Vitro Folding of Disulfide-Containing Globular Protein from Crude Bacterial Inclusion Bodies.

    Futami J, Tsushima Y, Tada H, *Seno M, Yamada H.

    J Biochem (Tokyo)   127 ( 3 )   435   2000.4

     More details

  • Stabilization of human RNase 1 by introduction of a disulfide bond between residues 4 and 118.

    J Biochem (Tokyo)   128 ( 2 )   245   2000.4

     More details

  • Convenient and efficient in vitro folding of disulfide-containing globular protein from crude bacterial inclusion bodies

    J Futami, Y Tsushima, H Tada, M Seno, H Yamada

    JOURNAL OF BIOCHEMISTRY   127 ( 3 )   435 - 441   2000.3

     More details

    Language:English   Publisher:JAPANESE BIOCHEMICAL SOC  

    We investigated how the folding yield of disulfide-containing globular proteins having positive net charges from crude bacterial inclusion bodies was affected by additives in the folding buffer. In screening folding conditions for human ribonucleases and its derivative, we found that addition of salt (about 0.4 M) to a folding buffer increased the folding yield. This suggested that electrostatic interaction between polyanionic impurities such as nucleic acids and cationic unfolded protein led to the formation of aggregates under the low-salt conditions. Since inclusion bodies were found to contain nucleic acids regardless of the electrostatic nature of the expressed protein, the electrostatic interaction between phosphate moieties of nucleic acids and basic amino acid residues of a denatured protein may be large enough to cause aggregation, and therefore the addition of salt in a folding buffer may generally be useful for promotion of protein folding from crude inclusion bodies. We further systematically investigated additives such as glycerol, guanidium chloride, and urea that are known to act as chemical chaperons, and found that these additives, together with salt, synergistically improved folding yield. This study, suggesting that the addition of salt into the folding buffer is one of the crucial points to be considered, may pave the way for a systematic investigation of the folding conditions of disulfide-containing foreign proteins from crude bacterial inclusion bodies.

    Web of Science

    researchmap

  • Cripto-1 induces apoptosis in HC-11 mouse mammary epithelial cells

    M. L. De Santis, I. Martinez-Lacaci, C. Bianco, M. Seno, B. Wallace-Jones, N. Kim, A. Ebert, C. Wechselberger, D. S. Salomon

    Cell Death and Differentiation   7 ( 2 )   189 - 196   2000

     More details

    Language:English   Publisher:Nature Publishing Group  

    Cripto-1 (CR-1) is an epidermal growth factor (EGF)-related protein. CR-1 can inhibit β-casein and whey acidic protein expression in mouse mammary epithelial cells. The present study demonstrates that CR-1 can induce apoptosis in HC-11 mouse mammary epithelial cells, as measured by bis-benzimide stained nuclei, TUNEL assay and cell death ELISA. Apoptosis could be observed after 2 days of exposure of confluent HC-11 cells to CR-1 in the absence of the survival factors EGF and insulin, with maximum apoptosis occurring at 3 days. A reduction in poly(ADP-ribose) polymerase (PARP) expression and an increase in β-catenin cleavage was found after 18 h of exposure to CR-1 suggesting that apoptosis was preceded by the induction of a caspase activity since the caspase inhibitor ZFAD.FMK could block the CR-1-induced reduction in PARP expression and CR-1-induced apoptosis. CR-1 was found to increase the expression of caspase-3-like protease. Although, the levels of p27(kip1) and p21Bax did not change after exposure to CR-1 for 18 h, the levels of Bcl-x(L) became undetectable. These studies suggest that CR-1 promotes apoptosis by mediating the induction of caspase-3-like protease and downregulating the expression of Bcl-xL.

    DOI: 10.1038/sj.cdd.4400588

    Scopus

    PubMed

    researchmap

  • Cripto-1 induces apoptosis in HC-11 mouse mammary epithelial cells

    M. L. De Santis, I. Martinez-Lacaci, C. Bianco, M. Seno, B. Wallace-Jones, N. Kim, A. Ebert, C. Wechselberger, D. S. Salomon

    Cell Death and Differentiation   7 ( 2 )   189 - 196   2000

     More details

    Language:English   Publisher:Nature Publishing Group  

    Cripto-1 (CR-1) is an epidermal growth factor (EGF)-related protein. CR-1 can inhibit β-casein and whey acidic protein expression in mouse mammary epithelial cells. The present study demonstrates that CR-1 can induce apoptosis in HC-11 mouse mammary epithelial cells, as measured by bis-benzimide stained nuclei, TUNEL assay and cell death ELISA. Apoptosis could be observed after 2 days of exposure of confluent HC-11 cells to CR-1 in the absence of the survival factors EGF and insulin, with maximum apoptosis occurring at 3 days. A reduction in poly(ADP-ribose) polymerase (PARP) expression and an increase in β-catenin cleavage was found after 18 h of exposure to CR-1 suggesting that apoptosis was preceded by the induction of a caspase activity since the caspase inhibitor ZFAD.FMK could block the CR-1-induced reduction in PARP expression and CR-1-induced apoptosis. CR-1 was found to increase the expression of caspase-3-like protease. Although, the levels of p27(kip1) and p21Bax did not change after exposure to CR-1 for 18 h, the levels of Bcl-x(L) became undetectable. These studies suggest that CR-1 promotes apoptosis by mediating the induction of caspase-3-like protease and downregulating the expression of Bcl-xL.

    DOI: 10.1038/sj.cdd.4400588

    Scopus

    PubMed

    researchmap

  • Immunohistochemical localization of betacellulin, a new member of the EGF family, in normal human pancreas and islet tumor cells

    Jun Ichiro Miyagawa, Toshiaki Hanafusa, Reiko Sasada, Koji Yamamoto, Koichi Igarashi, Katsumi Yamamori, Masaharu Seno, Hiroko Tada, Takao Nammo, Ming Li, Kazuya Yamagata, Hiromu Nakajima, Mitsuyoshi Namba, Masamichi Kuwajima, Yuji Matsuzawa

    Endocrine Journal   46 ( 6 )   755 - 764   1999.12

     More details

    Betacellulin (BTC) purified from mouse β cell tumor (βTC-3) is a new member of the epidermal growth factor (EGF) family which can bind receptor tyrosine kinase, EGF receptor (erbB1) and erbB4. It has been demonstrated that proBTC mRNA was abundantly expressed in human pancreas tissue, and that BTC converted amylase-secreting rat acinar cell line (AR42J) into insulin- secreting cells, suggesting that BTC might be important for the growth and/or differentiation of islet cells. However, the cell type producing BTC in the pancreas has not been clarified. In this study, we examined the localization of BTC in human pancreas and islet cell tumors. Immunohistochemistry using specific antibodies to human BTC revealed that this protein was produced in a cells and duct cells, and probably in β cells in normal adult pancreas. Furthermore, strong immunoreactivity to BTC was detected in primitive duct cells of the fetal pancreas, and both insulinoma and glucagonoma cells also showed positive immunoreactivity to BTC. EGF receptor (erbB1) and erbB4 were expressed mainly in islet and duet cells, and duct cells, respectively. These results demonstrate the localization of BTC and its receptors, and suggest that BTC may be one of the factors that have physiologically important roles such as growth and differentiation of islet cells in the human pancreas.

    DOI: 10.1507/endocrj.46.755

    Scopus

    PubMed

    researchmap

  • Immunohistochemical localization of betacellulin, a new member of the EGF family, in normal human pancreas and islet tumor cells

    J Miyagawa, T Hanafusa, R Sasada, K Yamamoto, K Igarashi, K Yamamori, M Seno, H Tada, T Nammo, M Li, K Yamagata, H Nakajima, M Namba, M Kuwajima, Y Matsuzawa

    ENDOCRINE JOURNAL   46 ( 6 )   755 - 764   1999.12

     More details

    Language:English   Publisher:JAPAN ENDOCRINE SOCIETY  

    Betacellulin (BTC) purified from mouse beta cell tumor (beta TC-3) is a new member of the epidermal growth factor (EGF) family which can bind receptor tyrosine kinase, EGF receptor (erbB1) and erbB4. It has been demonstrated that proBTC mRNA was abundantly expressed in human pancreas tissue, and that ETC converted amylase-secreting rat acinar cell line (AR42J) into insulin-secreting cells, suggesting that ETC might be important for the growth and/or differentiation of islet cells. However, the cell type producing ETC in the pancreas has not been clarified. In this study, we examined the localization of ETC in human pancreas and islet cell tumors. Immunohistochemistry using specific antibodies to human ETC revealed that this protein was produced in alpha cells and duct cells, and probably in beta cells in normal adult pancreas. Furthermore, strong immunoreactivity to ETC was detected in primitive duct cells of the fetal pancreas, and both insulinoma and glucagonoma cells also showed positive immunoreactivity to ETC. EGF receptor (erbB1) and erbB4 were expressed mainly in islet and duct cells, and duct cells, respectively. These results demonstrate the localization of ETC and its receptors, and suggest that ETC may be one of the factors that have physiologically important roles such as growth and differentiation of islet cells in the human pancreas.

    DOI: 10.1507/endocrj.46.755

    Web of Science

    researchmap

  • Immunohistochemical localization of betacellulin, a new member of the EGF family, in normal human pancreas and islet tumor cells

    Jun Ichiro Miyagawa, Toshiaki Hanafusa, Reiko Sasada, Koji Yamamoto, Koichi Igarashi, Katsumi Yamamori, Masaharu Seno, Hiroko Tada, Takao Nammo, Ming Li, Kazuya Yamagata, Hiromu Nakajima, Mitsuyoshi Namba, Masamichi Kuwajima, Yuji Matsuzawa

    Endocrine Journal   46 ( 6 )   755 - 764   1999.12

     More details

    Betacellulin (BTC) purified from mouse β cell tumor (βTC-3) is a new member of the epidermal growth factor (EGF) family which can bind receptor tyrosine kinase, EGF receptor (erbB1) and erbB4. It has been demonstrated that proBTC mRNA was abundantly expressed in human pancreas tissue, and that BTC converted amylase-secreting rat acinar cell line (AR42J) into insulin- secreting cells, suggesting that BTC might be important for the growth and/or differentiation of islet cells. However, the cell type producing BTC in the pancreas has not been clarified. In this study, we examined the localization of BTC in human pancreas and islet cell tumors. Immunohistochemistry using specific antibodies to human BTC revealed that this protein was produced in a cells and duct cells, and probably in β cells in normal adult pancreas. Furthermore, strong immunoreactivity to BTC was detected in primitive duct cells of the fetal pancreas, and both insulinoma and glucagonoma cells also showed positive immunoreactivity to BTC. EGF receptor (erbB1) and erbB4 were expressed mainly in islet and duet cells, and duct cells, respectively. These results demonstrate the localization of BTC and its receptors, and suggest that BTC may be one of the factors that have physiologically important roles such as growth and differentiation of islet cells in the human pancreas.

    DOI: 10.1507/endocrj.46.755

    Scopus

    PubMed

    researchmap

  • Inhibition of cell growth by a fused protein of human ribonuclease 1 and human basic fibroblast growth factor

    J Futami, M Seno, M Ueda, H Tada, H Yamada

    PROTEIN ENGINEERING   12 ( 11 )   1013 - 1019   1999.11

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    Pancreatic-type RNases are considered to have cytotoxic potential due to their ability to degrade RNA molecules when they enter the cytosol. However, most of these RNases show little cytotoxicity because cells have no active uptake mechanism for these RNases and because the ubiquitous cytoplasmic RNase inhibitor is considered to play a protective role against the endocytotic leak of RNases from the outside of cells. To study the cytotoxic potential of RNase toward malignant cells targeting growth factor receptors, the C-terminus of human RNase 1 was fused to the N-terminus of human basic fibroblast growth factor (bFGF). This RNase-FGF fused protein effectively inhibited the growth of mouse melanoma cell line B16/BL6 with high levels of cell surface FGF receptor. This effect appeared to result from prolongation of the overall cell cycle rather than the killing of cells or specific arrest in a particular phase of the cell cycle. Thus, human RNase 1 fused to a ligand of cell surface molecules, such as the FGF receptor, is shown to be an effective candidate for a selective cell targeting agent with low toxic effects on normal cell types.

    Web of Science

    researchmap

  • Cripto-1 induces phosphatidylinositol 3 &apos;-kinase-dependent phosphorylation of AKT and glycogen synthase kinase 3 beta in human cervical carcinoma cells

    AD Ebert, C Wechselberger, S Frank, B Wallace-Jones, M Seno, Martinez-Lacaci, I, C Bianco, M De Santis, HK Weitzel, DS Salomon

    CANCER RESEARCH   59 ( 18 )   4502 - 4505   1999.9

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    Cripto-1 (CR-1), a member of the epidermal growth factor-CFC peptide family, activates the ras/raf/mitogen-activated protein/extracellular signal-regulated kinase/mitogen-activated protein kinase pathway. In the present study, the role of CR-1 in the phosphatidylinositol 3&apos;-kinase (PI3K)/AKT (protein kinase B)/glycogen synthase kinase 3 beta (GSK-3 beta)dependent signaling pathway was evaluated in human SiHa cervical carcinoma cells. Our data demonstrate that CR-1 can enhance the tyrosine phosphorylation of the p85 regulatory subunit of PI3K and transiently induce the phosphorylation of AKT in a time- and dose-dependent manner. In addition, CR-1 was found to induce the phosphorylation of GSK-3 beta. Phosphorylation of AKT and GSK-3 beta by CR-1 can be blocked by LY294002, a specific inhibitor of PI3K;, thus leading to apoptosis. Finally, the apoptotic effect of LY294002 can be partially rescued by exogenous CR-1. In summary, our data suggest that human CR-1 may function as a survival factor through a PI3K-dependent signaling pathway involving AKT and GSK-3 beta.

    Web of Science

    researchmap

  • Cripto-1 induces phosphatidylinositol 3''-kinase-dependent phosphorylation of AKT and glycogen synthase kinase 3beta in human cervical carcinoma cells.

    Ebert AD, Wechselberger C, Frank S, Wallace-Jones B, *Seno M, Martinez-LacaciI, Bianco C, De Santis M, Weitzel HK, Salomon DS.

    Cancer Res.   59 ( 18 )   4502   1999.4

     More details

  • Inhibition of Cell Growth by a fused Protein of Human Basic Fibroblast Growth Factor.

    Protein Engineering   12 ( 11 )   1013   1999.4

     More details

  • Cripto-1 induces phosphatidylinositol 3''-kinase-dependent phosphorylation of AKT and glycogen synthase kinase 3beta in human cervical carcinoma cells.

    Cancer Res.   59 ( 18 )   4502   1999.4

     More details

  • Inhibition of Cell Growth by a fused Protein of Human Basic Fibroblast Growth Factor.

    Futami J, *Seno M, Ueda M, Tada H, Yamada H

    Protein Engineering   12 ( 11 )   1013   1999.4

     More details

  • Cripto-1 indirectly stimulates the tyrosine phosphorylation of erb B-4 through a novel receptor

    Caterina Bianco, Subha Kannan, Marta De Santis, Masaharu Seno, Careen K. Tang, Isabel Martinez-Lacaci, Nancy Kim, Brenda Wallace-Jones, Marc E. Lippman, Andreas D. Ebert, Christian Wechselberger, David S. Salomon

    Journal of Biological Chemistry   274 ( 13 )   8624 - 8629   1999.3

     More details

    Language:English  

    Cripto-1 (CR-1) is a recently discovered protein of the epidermal growth factor family that fails to directly bind to any of the four known erb B type 1 receptor tyrosine kinases. The present study demonstrates that CR-1 indirectly induces tyrosine phosphorylation of erb B-4 but not of the epidermal growth factor-related receptors erb B-2 and erb B-3 in different mouse and human mammary epithelial cell lines. In addition, down-regulation of erb B-4 in NMuMG mouse mammary epithelial cells and in T47D human breast cancer cells, using an anti-orb B-4 blocking antibody or a hammerhead ribozyme vector targeted to orb B-4 mRNA, impairs the ability of CR-1 to fully activate mitogen-activated protein kinase. Finally, chemical cross- linking of 125I-CR-1 to mouse and human mammary epithelial cell membranes results in the labeling of two specific bands with a molecular weight of 130 and 60 kDa, suggesting that the CR-1 receptor represents a novel receptor structurally unrelated to any of the known type I receptor tyrosine kinases. In conclusion, these data demonstrate that CR-1, upon binding to an unknown receptor, can enhance the tyrosine kinase activity of erb B-4 and that a functional erb B-4 receptor is required for CR-1-induced MAPK activation.

    DOI: 10.1074/jbc.274.13.8624

    Scopus

    PubMed

    researchmap

  • Cripto-1 indirectly stimulates the tyrosine phosphorylation of erb B-4 through a novel receptor

    Caterina Bianco, Subha Kannan, Marta De Santis, Masaharu Seno, Careen K. Tang, Isabel Martinez-Lacaci, Nancy Kim, Brenda Wallace-Jones, Marc E. Lippman, Andreas D. Ebert, Christian Wechselberger, David S. Salomon

    Journal of Biological Chemistry   274 ( 13 )   8624 - 8629   1999.3

     More details

    Language:English  

    Cripto-1 (CR-1) is a recently discovered protein of the epidermal growth factor family that fails to directly bind to any of the four known erb B type 1 receptor tyrosine kinases. The present study demonstrates that CR-1 indirectly induces tyrosine phosphorylation of erb B-4 but not of the epidermal growth factor-related receptors erb B-2 and erb B-3 in different mouse and human mammary epithelial cell lines. In addition, down-regulation of erb B-4 in NMuMG mouse mammary epithelial cells and in T47D human breast cancer cells, using an anti-orb B-4 blocking antibody or a hammerhead ribozyme vector targeted to orb B-4 mRNA, impairs the ability of CR-1 to fully activate mitogen-activated protein kinase. Finally, chemical cross- linking of 125I-CR-1 to mouse and human mammary epithelial cell membranes results in the labeling of two specific bands with a molecular weight of 130 and 60 kDa, suggesting that the CR-1 receptor represents a novel receptor structurally unrelated to any of the known type I receptor tyrosine kinases. In conclusion, these data demonstrate that CR-1, upon binding to an unknown receptor, can enhance the tyrosine kinase activity of erb B-4 and that a functional erb B-4 receptor is required for CR-1-induced MAPK activation.

    DOI: 10.1074/jbc.274.13.8624

    Scopus

    PubMed

    researchmap

  • Cripto-1 indirectly stimulates the tyrosine phosphorylation of erb B-4 through a novel receptor

    C Bianco, S Kannan, M De Santis, M Seno, CK Tang, Martinez-Lacaci, I, N Kim, B Wallace-Jones, ME Lippman, AD Ebert, C Wechselberger, DS Salomon

    JOURNAL OF BIOLOGICAL CHEMISTRY   274 ( 13 )   8624 - 8629   1999.3

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Cripto-1 (CR-1) is a recently discovered protein of the epidermal growth factor family that fails to directly bind to any of the four known erb B type 1 receptor tyrosine kinases. The present study demonstrates that CR-1 indirectly induces tyrosine phosphorylation of erb B-4 but not of the epidermal growth factor-related receptors erb B-2 and erb B-3 in different mouse and human mammary epithelial cell lines. In addition, downregulation of erb B-4 in NMuMG mouse mammary epithelial cells and in T47D human breast cancer cells, using an anti-erb B-4 blocking antibody or a hammerhead ribozyme vector targeted to erb B-4 mRNA, impairs the ability of CR-1 to fully activate mitogen-activated protein kinase. Finally, chemical cross-linking of I-125-CR-1 to mouse and human mammary epithelial cell membranes results in the labeling of two specific bands with a molecular weight of 130 and 60 kDa, suggesting that the CR-1 receptor represents a novel receptor structurally unrelated to any of the known type I receptor tyrosine kinases. In conclusion, these data demonstrate that CR-1, upon binding to an unknown receptor, can enhance the tyrosine kinase activity of erb B-4 and that a functional erb B-4 receptor is required for CR-1-induced MAPK activation.

    DOI: 10.1074/jbc.274.13.8624

    Web of Science

    researchmap

  • Processing and juxtacrine activity of membrane-anchored betacellulin

    H Tada, R Sasada, Y Kawaguchi, Kojima, I, WJ Gullick, DS Salomon, K Igarashi, M Seno, H Yamada

    JOURNAL OF CELLULAR BIOCHEMISTRY   72 ( 3 )   423 - 434   1999.3

     More details

    Language:English   Publisher:WILEY-LISS  

    Betacellulin (BTC) was originally isolated as a secreted growth factor from a mouse pancreatic beta-tumor cell line, whereas the cDNA sequence predicts that ETC is synthesized as a larger transmembrane protein. In the present study, we have characterized the membrane-anchored forms of BTC, using Chinese hamster ovary (CHO) cells, mouse fibroblast A9 cells, and a human breast cancer cell line MCF-7, all of which were stably transfected with human ETC cDNA. A9 and MCF-7 transfectants produced membrane-anchored ETC isoforms of 21, 25, 29, and 40 kDa on the cell surface, as well as a secreted BTC isoform. CHO transfectants secreted little ETC but accumulated the membrane-anchored isoforms. The cleavage of the membrane-anchored forms to release a secreted from of ETC was not enhanced by biological mediators such as a phorbol ester, which stimulates the cleavage of other membrane-anchored growth factors. The membrane-anchored forms of ETC expressed on the transfected cells induced the insulin production and/or promoted the growth in subclones of AR42J rat pancreatic cells. These results suggest that the membrane-anchored ETC can function as a juxtacrine factor in regulating the growth and differentiation of pancreatic endocrine cells. J. Cell. Biochem. 72.423-134, 1999. (C) 1999 Wiley-Liss, Inc.

    DOI: 10.1002/(SICI)1097-4644(19990301)72:3<423::AID-JCB11>3.0.CO;2-P

    Web of Science

    researchmap

  • Genes expressed during the differentiation of pancreatic ARA2J cells into insulin-secreting cells

    H Mashima, S Yamada, T Tajima, M Seno, H Yamada, J Takeda, Kojima, I

    DIABETES   48 ( 2 )   304 - 309   1999.2

     More details

    Language:English   Publisher:AMER DIABETES ASSOC  

    dPancreatic AR42J cells have the feature of pluripotency of the common precursor cells of the pancreas. Dexamethasone (Dx) converts them to exocrine cells, whereas activin A (Act) converts them into endocrine cells expressing pancreatic polypeptide. A combination of Act and betacellulin (BTC) converts them further into insulin-secreting cells, The present study identifies some of the genes involved in the process of differentiation that is induced by these factors, using the mRNA differential display and screening of the cDNA expression array. The expression levels of 7 genes were increased by Act alone, and a combination of Act and ETC increased the expression of 25 more genes. Of these, 16 represented known genes or homologues of genes characterized previously, Nine of the identified genes were unrelated to any other sequences in the database. An inhibitor of the mitogen-activated protein kinase pathway, PD098059, which blocks the differentiation into insulin-secreting cells, inhibited the expression of 18 of the 25 genes, suggesting that the proteins encoded by these genes are associated with the differentiation into insulin-producing cells, These include known genes encoding extracellular signaling molecules, such as parathyroid hormone-related peptide, cyloskeletal proteins, and intracellular signaling molecules. Identification and characterization of these differentially expressed genes should help to clarify the molecular mechanism of differentiation of pancreatic cells and the; gene products that enable the beta-cells to produce insulin.

    DOI: 10.2337/diabetes.48.2.304

    Web of Science

    researchmap

  • The three-dimensional structure of human RNase 4, unliganded and complexed with d(Up), reveals the basis for its uridine selectivity

    SS Terzyan, R Peracaula, R de Llorens, Y Tsushima, H Yamada, M Seno, FX Gomis-Ruth, M Coll

    JOURNAL OF MOLECULAR BIOLOGY   285 ( 1 )   205 - 214   1999.1

     More details

    Language:English   Publisher:ACADEMIC PRESS LTD  

    The RNase 4 family is unique among RNase enzymes, displaying the highest level of sequence similarity and encompassing the shortest polypeptide chain. It is the only one showing high specificity. The human representative is an intracellular and plasma enzyme, first isolated from colon adenocarcinoma cell line HT-29. The crystal structures of human recombinant RNase 4, unliganded and in complex with d(Up), have been determined, revealing in the unique active site an explanation for the uridine specificity. Arg101, at a position not involved in catalysis in the other RNase enzymes, penetrates the enzyme moiety shaping the recognition pocket, a flip that is mediated by the interaction with the (shorter chain) C-terminal carboxylate group, providing an anchoring point for the O4 atom of the substrate uridine. The bulky Phe42 side-chain forces Asp80 to be in the chi(1) = -72.49 degrees rotamer, accepting a hydrogen bond from Thr44, further converting the latter into a hydrogen bond acceptor. This favours an interaction with the -NH-donor group of uridine at position 3 over that with the =N-acceptor of cytidine. The two chemical groups that distinguish uracyl from cytosine are used by the enzyme to discriminate between these two bases. (C) 1999 Academic Press.

    DOI: 10.1006/jmbi.1998.2288

    Web of Science

    researchmap

  • Genes expressed during the differentiation of pancreatic AR42J cells into insulin-secreting cells

    Hirosato Mashima, Shirou Yamada, Tomoko Tajima, Masaharu Seno, Hidenori Yamada, Jun Takeda, Itaru Kojima

    Diabetes   48 ( 2 )   304 - 309   1999

     More details

    Language:English   Publisher:American Diabetes Association Inc.  

    Pancreatic AR42J cells have the feature of pluripotency of the common precursor cells of the pancreas. Dexamethasone (Dx) converts them to exocrine cells, whereas activin A (Act) converts them into endocrine cells expressing pancreatic polypeptide. A combination of Act and betacellulin (BTC) converts them further into insulin-secreting cells. The present study identifies some of the genes involved in the process of differentiation that is induced by these factors, using the mRNA differential display and screening of the cDNA expression array. The expression levels of 7 genes were increased by Act alone, and a combination of Act and BTC increased the expression of 25 more genes. Of these, 16 represented known genes or homologues of genes characterized previously. Nine of the identified genes were unrelated to any other sequences in the database. An inhibitor of the mitogen-activated protein kinase pathway, PD098059, which blocks the differentiation into insulin-secreting cells, inhibited the expression of 18 of the 25 genes, suggesting that the proteins encoded by these genes are associated with the differentiation into insulin-producing cells. These include known genes encoding extracellular signaling molecules, such as parathyroid hormone- related peptide, cytoskeletal proteins, and intracellular signaling molecules. Identification and characterization of these differentially expressed genes should help to clarify the molecular mechanism of differentiation of pancreatic cells and the gene products that enable the β- cells to produce insulin.

    DOI: 10.2337/diabetes.48.2.304

    Scopus

    PubMed

    researchmap

  • Genes expressed during the differentiation of pancreatic AR42J cells into insulin-secreting cells

    Hirosato Mashima, Shirou Yamada, Tomoko Tajima, Masaharu Seno, Hidenori Yamada, Jun Takeda, Itaru Kojima

    Diabetes   48 ( 2 )   304 - 309   1999

     More details

    Language:English   Publisher:American Diabetes Association Inc.  

    Pancreatic AR42J cells have the feature of pluripotency of the common precursor cells of the pancreas. Dexamethasone (Dx) converts them to exocrine cells, whereas activin A (Act) converts them into endocrine cells expressing pancreatic polypeptide. A combination of Act and betacellulin (BTC) converts them further into insulin-secreting cells. The present study identifies some of the genes involved in the process of differentiation that is induced by these factors, using the mRNA differential display and screening of the cDNA expression array. The expression levels of 7 genes were increased by Act alone, and a combination of Act and BTC increased the expression of 25 more genes. Of these, 16 represented known genes or homologues of genes characterized previously. Nine of the identified genes were unrelated to any other sequences in the database. An inhibitor of the mitogen-activated protein kinase pathway, PD098059, which blocks the differentiation into insulin-secreting cells, inhibited the expression of 18 of the 25 genes, suggesting that the proteins encoded by these genes are associated with the differentiation into insulin-producing cells. These include known genes encoding extracellular signaling molecules, such as parathyroid hormone- related peptide, cytoskeletal proteins, and intracellular signaling molecules. Identification and characterization of these differentially expressed genes should help to clarify the molecular mechanism of differentiation of pancreatic cells and the gene products that enable the β- cells to produce insulin.

    DOI: 10.2337/diabetes.48.2.304

    Scopus

    PubMed

    researchmap

  • 新しい臓器特異的癌関連遺伝子のクローニングと癌遺伝子産物を分子標的とした治療法の開発

    上田 政和, 菊池 潔, 板野 理, プサラス・キリヤコス, 北島 政樹, 妹尾 昌治

    日本癌治療学会誌   33 ( 3 )   355 - 355   1998.8

     More details

    Language:Japanese   Publisher:(一社)日本癌治療学会  

    researchmap

  • Identification of the genes associated with differentiation of AR42J cells into insulin-secreting cells

    H Mashima, J Takeda, M Seno, H Yamada, Kojima, I

    DIABETES   47   A176 - A176   1998.5

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER DIABETES ASSOC  

    Web of Science

    researchmap

  • Molecular targeting for activated T cell by recombinant human RNase and IL-2.

    M Ueda, K Psarras, M Tanabe, T Yamamura, M Kitajima, M Seno, S Komatsu

    GASTROENTEROLOGY   114 ( 4 )   A1186 - A1186   1998.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:W B SAUNDERS CO  

    Web of Science

    researchmap

  • Multiple-labeling of oligonucleotide probes for in situ hybridization

    Junzo Sasaki, Hitoshi Yamamoto, Takako Nomura, Junko Matsuura, Masaharu Seno, Eisuke F. Sato, Masayasu Inoue

    Acta Histochemica et Cytochemica   31 ( 4 )   275 - 279   1998

     More details

    Language:English   Publisher:Japan Society of Histochemistry and Cytochemistry  

    We describe here a method to synthesize probes for in situ hybridization. This method provides more efficient incorporation of the reporter molecules such as 35S-UTP or digoxigenin-UTP into the oligonucleotide probes than other methods. Two 99-base oligonucleotides complementary to each other were obtained as purified and lyophilized products (&gt
    99%). These oligonucleotides were designed as follows. The sequence of 77 bases derived from reported cDNA sequence in the literature was flanked by the restriction sites of EcoR I and Hind III (6 bases for each) with extended random sequences of 5 bases at both ends (total 99 bases). Both oligonucleotides were then annealed and digested with EcoR I and Hind III. The gel-purified EcoR I/Hind III-cut DNA fragment was cloned into the pGEM4Z vector. The resultant plasmid DNA was linearized with EcoR I or Hind III and used as a template for the synthesis of labeled sense or antisense riboprobes. The amelogenin probes prepared by this method clearly distinguished the localized expression of mRNA when applied to in situ hybridization.

    DOI: 10.1267/ahc.31.275

    Scopus

    researchmap

  • BALB/c 3T3 cells co-expressing FGF-2 and soluble FGF receptor acquire tumorigenicity

    Masaharu Seno, Akinori Masago, Atsushi Nishimura, Hiroko Tada, Megumi Kosaka, Reiko Sasada, Koichi Igarashi, Satimaru Seno, Hidenori Yamada

    Cytokine   10 ( 4 )   290 - 294   1998

     More details

    Language:English   Publisher:Academic Press  

    The physiological significance of the soluble fibroblast growth factor (FGF) receptors is not clear yet although they are present in blood, vitreous fluid and in the extracellular matrix of vascular endothelial cells. A hypothesis that they might help FGF-2 release from cells is very interesting because FGF-2 does not have clear secretion signal and the mechanism of the secretion of FGF-2 is still unclear. Single overexpression of FGF-2 is related neither to the secretion potential of the molecule nor to the tumorigenicity of the cells. In this report, BALB/c 3T3 cells transformed with the full length of human FGF-2 cDNA are further transformed with the cDNA coding the extracellular domain of human FGF receptor 1. The obtained transformants co-expressing FGF-2 and soluble FGF receptor are highly tumorigenic in nude mice, while the parental cells do not show any tumorigenicity. In the conditioned medium of the double-transformants, FGF-2 is immunologically detected. These results suggest that naturally produced soluble form of FGF receptor supports the release of FGF-2 from the cells and that over-expression of these two molecules leads to induce the malignant tumours in vivo.

    DOI: 10.1006/cyto.1997.0286

    Scopus

    PubMed

    researchmap

  • BALB/c 3T3 cells co-expressing FGF-2 and soluble FGF receptor acquire tumorigenicity

    Masaharu Seno, Akinori Masago, Atsushi Nishimura, Hiroko Tada, Megumi Kosaka, Reiko Sasada, Koichi Igarashi, Satimaru Seno, Hidenori Yamada

    Cytokine   10 ( 4 )   290 - 294   1998

     More details

    Language:English   Publisher:Academic Press  

    The physiological significance of the soluble fibroblast growth factor (FGF) receptors is not clear yet although they are present in blood, vitreous fluid and in the extracellular matrix of vascular endothelial cells. A hypothesis that they might help FGF-2 release from cells is very interesting because FGF-2 does not have clear secretion signal and the mechanism of the secretion of FGF-2 is still unclear. Single overexpression of FGF-2 is related neither to the secretion potential of the molecule nor to the tumorigenicity of the cells. In this report, BALB/c 3T3 cells transformed with the full length of human FGF-2 cDNA are further transformed with the cDNA coding the extracellular domain of human FGF receptor 1. The obtained transformants co-expressing FGF-2 and soluble FGF receptor are highly tumorigenic in nude mice, while the parental cells do not show any tumorigenicity. In the conditioned medium of the double-transformants, FGF-2 is immunologically detected. These results suggest that naturally produced soluble form of FGF receptor supports the release of FGF-2 from the cells and that over-expression of these two molecules leads to induce the malignant tumours in vivo.

    DOI: 10.1006/cyto.1997.0286

    Scopus