2021/04/15 更新

写真a

エグチ タカノリ
江口 傑徳
EGUCHI Takanori
所属
医歯薬学域 助教
職名
助教
通称等の別名
たかのり
プロフィール

江口 傑徳(たかのり)は、日本の生命科学者、教育者。分子機能や細胞間コミュニケーションの視点から生体を理解し、新しい医療の創出を目指している。これまでに50編以上の論文や著書を発表し、代表論文は、

Organoids with cancer stem cell-like properties secrete exosomes and HSP90 in a 3D nanoenvironment. Eguchi T et al, PLoS One 13 (2), e0191109

Triple knockdown of CDC37, HSP90-alpha and HSP90-beta diminishes extracellular vesicles-driven malignancy events and macrophage M2 polarization in oral cancer. Ono K et al, Journal of Extracellular Vesicles 9 (1), 1769373

Novel transcription factor-like function of human matrix metalloproteinase 3 regulating the CTGF/CCN2 gene. Eguchi T et al, Molecular and Cellular Biology 28 (7), 2391-2413(日本軟骨代謝学会賞、International CCN Society Springer Scholarshipなどを受賞)

略歴:岡山大学大学院修了・歯学博士(2003)、日本学術振興会特別研究員、国立長寿医療研究センター室長、ハーバード大学医学大学院(2011-2015)を経て、現職。

学位

  • 博士(歯学) ( 2003年3月   岡山大学 )

研究キーワード

  • 非コードRNA

  • 遺伝子制御

  • オミクス

  • 細胞外小胞

  • Tumoroids

  • 三次元培養システムズ

  • 細胞分子生物学

  • ムーンライティングタンパク質

  • 細胞間コミュニケーション

経歴

  • ハーバード大学医学大学院   インストラクター

    2011年9月 - 2015年12月

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  • 国立研究開発法人国立長寿医療研究センター   室長

    2009年1月 - 2011年6月

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  • 岡山大学

    2021年4月 - 現在

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  • 岡山大学大学院医歯薬学研究科   助教

    2016年4月 - 現在

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  • ハーバード大学医学大学院   リサーチフェロー

    2011年7月 - 2011年8月

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  • 国立長寿医療センター研究所   流動研究員

    2007年9月 - 2008年12月

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  • 日本学術振興会   特別研究員(PD)

    2003年4月 - 2006年3月

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▼全件表示

所属学協会

▼全件表示

委員歴

  • 日本臨床ストレス応答学会   幹事  

    2020年4月   

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    団体区分:学協会

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  • 日本臨床ストレス応答学会   評議員  

    2016年 - 現在   

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    団体区分:学協会

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論文

  • Triple knockdown of CDC37, HSP90-alpha and HSP90-beta diminishes extracellular vesicles-driven malignancy events and macrophage M2 polarization in oral cancer. 国際誌

    Kisho Ono, Chiharu Sogawa, Hotaka Kawai, Manh Tien Tran, Eman A Taha, Yanyin Lu, May Wathone Oo, Yuka Okusha, Hirohiko Okamura, Soichiro Ibaragi, Masaharu Takigawa, Ken-Ichi Kozaki, Hitoshi Nagatsuka, Akira Sasaki, Kuniaki Okamoto, Stuart K Calderwood, Takanori Eguchi

    Journal of extracellular vesicles9 ( 1 ) 1769373 - 1769373   2020年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Evidence has been accumulating to indicate that extracellular vesicles (EVs), including exosomes, released by cancer cells can foster tumour progression. The molecular chaperones - CDC37, HSP90α and HSP90β play key roles in cancer progression including epithelial-mesenchymal transition (EMT), although their contribution to EVs-mediated cell-cell communication in tumour microenvironment has not been thoroughly examined. Here we show that triple depletion of the chaperone trio attenuates numerous cancer malignancy events exerted through EV release. Metastatic oral cancer-derived EVs (MEV) were enriched with HSP90α HSP90β and cancer-initiating cell marker CD326/EpCAM. Depletion of these chaperones individually induced compensatory increases in the other chaperones, whereas triple siRNA targeting of these molecules markedly diminished the levels of the chaperone trio and attenuated EMT. MEV were potent agents in initiating EMT in normal epithelial cells, a process that was attenuated by the triple chaperone depletion. The migration, invasion, and in vitro tumour initiation of oral cancer cells were significantly promoted by MEV, while triple depletion of CDC37/HSP90α/β reversed these MEV-driven malignancy events. In metastatic oral cancer patient-derived tumours, HSP90β was significantly accumulated in infiltrating tumour-associated macrophages (TAM) as compared to lower grade oral cancer cases. HSP90-enriched MEV-induced TAM polarization to an M2 phenotype, a transition known to support cancer progression, whereas the triple chaperone depletion attenuated this effect. Mechanistically, the triple chaperone depletion in metastatic oral cancer cells effectively reduced MEV transmission into macrophages. Hence, siRNA-mediated knockdown of the chaperone trio (CDC37/HSP90α/HSP90β) could potentially be a novel therapeutic strategy to attenuate several EV-driven malignancy events in the tumour microenvironment. Abbreviations: CDC37: cell division control 37; EMT: epithelial-mesenchymal transmission; EV: extracellular vesicles; HNSCC: head and neck squamous cell carcinoma; HSP90: heat shock protein 90; TAM: tumour-associated macrophage.

    DOI: 10.1080/20013078.2020.1769373

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  • Knockout of MMP3 Weakens Solid Tumor Organoids and Cancer Extracellular Vesicles. 国際誌

    Eman A Taha, Chiharu Sogawa, Yuka Okusha, Hotaka Kawai, May Wathone Oo, Abdellatif Elseoudi, Yanyin Lu, Hitoshi Nagatsuka, Satoshi Kubota, Ayano Satoh, Kuniaki Okamoto, Takanori Eguchi

    Cancers12 ( 5 )   2020年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The tumor organoid (tumoroid) model in three-dimensional (3D) culture systems has been developed to reflect more closely the in vivo tumors than 2D-cultured tumor cells. Notably, extracellular vesicles (EVs) are efficiently collectible from the culture supernatant of gel-free tumoroids. Matrix metalloproteinase (MMP) 3 is a multi-functional factor playing crucial roles in tumor progression. However, roles of MMP3 within tumor growth and EVs have not unveiled. Here, we investigated the protumorigenic roles of MMP3 on integrities of tumoroids and EVs. We generated MMP3-knockout (KO) cells using the CRISPR/Cas9 system from rapidly metastatic LuM1 tumor cells. Moreover, we established fluorescent cell lines with palmitoylation signal-fused fluorescent proteins (tdTomato and enhanced GFP). Then we confirmed the exchange of EVs between cellular populations and tumoroids. LuM1-tumoroids released large EVs (200-1000 nm) and small EVs (50-200 nm) while the knockout of MMP3 resulted in the additional release of broken EVs from tumoroids. The loss of MMP3 led to a significant reduction in tumoroid size and the development of the necrotic area within tumoroids. MMP3 and CD9 (a category-1 EV marker tetraspanin protein) were significantly down-regulated in MMP3-KO cells and their EV fraction. Moreover, CD63, another member of the tetraspanin family, was significantly reduced only in the EVs fractions of the MMP3-KO cells compared to their counterpart. These weakened phenotypes of MMP3-KO were markedly rescued by the addition of MMP3-rich EVs or conditioned medium (CM) collected from LuM1-tumoroids, which caused a dramatic rise in the expression of MMP3, CD9, and Ki-67 (a marker of proliferating cells) in the MMP3-null/CD9-low tumoroids. Notably, MMP3 enriched in tumoroids-derived EVs and CM deeply penetrated recipient MMP3-KO tumoroids, resulting in a remarkable enlargement of solid tumoroids, while MMP3-null EVs did not. These data demonstrate that EVs can mediate molecular transfer of MMP3, resulting in increasing the proliferation and tumorigenesis, indicating crucial roles of MMP3 in tumor progression.

    DOI: 10.3390/cancers12051260

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  • Antiparkinson Drug Benztropine Suppresses Tumor Growth, Circulating Tumor Cells, and Metastasis by Acting on SLC6A3/DAT and Reducing STAT3. 国際誌

    Chiharu Sogawa, Takanori Eguchi, Manh Tien Tran, Masayuki Ishige, Kilian Trin, Yuka Okusha, Eman Ahmed Taha, Yanyin Lu, Hotaka Kawai, Norio Sogawa, Masaharu Takigawa, Stuart K Calderwood, Kuniaki Okamoto, Ken-Ichi Kozaki

    Cancers12 ( 2 )   2020年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Tumor growth, progression, and therapy resistance are crucial factors in the prognosis of cancer. The properties of three-dimensional (3D) tumor-like organoids (tumoroids) more closely resemble in vivo tumors compared to two-dimensionally cultured cells and are therefore effectively used for assays and drug screening. We here established a repurposed drug for novel anticancer research and therapeutics using a 3D tumoroid-based screening system. We screened six pharmacologically active compounds by using an original tumoroid-based multiplex phenotypic screening system with a matrix metalloproteinase 9 (MMP9) promoter-driven fluorescence reporter for the evaluation of both tumoroid formation and progression. The antiparkinson drug benztropine was the most effective compound uncovered by the screen. Benztropine significantly inhibited in vitro tumoroid formation, cancer cell survival, and MMP9 promoter activity. Benztropine also reduced the activity of oncogenic signaling transducers and trans-activators for MMP9, including STAT3, NF-κB, and β-catenin, and the properties of cancer stem cells/cancer-initiating cells. Benztropine and GBR-12935 directly targeted the dopamine transporter DAT/SLC6A3, whose genetic alterations such as amplification were correlated with poor prognosis for cancer patients. Benztropine also inhibited the tumor growth, circulating tumor cell (CTC) number, and rate of metastasis in a tumor allograft model in mice. In conclusion, we propose the repurposing of benztropine for anticancer research and therapeutics that can suppress tumor progression, CTC, and metastasis of aggressive cancers by reducing key pro-tumorigenic factors.

    DOI: 10.3390/cancers12020523

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  • Organoids with cancer stem cell-like properties secrete exosomes and HSP90 in a 3D nanoenvironment. 査読 国際誌

    Takanori Eguchi, Chiharu Sogawa, Yuka Okusha, Kenta Uchibe, Ryosuke Iinuma, Kisho Ono, Keisuke Nakano, Jun Murakami, Manabu Itoh, Kazuya Arai, Toshifumi Fujiwara, Yuri Namba, Yoshiki Murata, Kazumi Ohyama, Manami Shimomura, Hirohiko Okamura, Masaharu Takigawa, Tetsuya Nakatsura, Ken-Ichi Kozaki, Kuniaki Okamoto, Stuart K Calderwood

    PloS one13 ( 2 ) e0191109   2018年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Public Library of Science  

    Ability to form cellular aggregations such as tumorspheres and spheroids have been used as a morphological marker of malignant cancer cells and in particular cancer stem cells (CSC). However, the common definition of the types of cellular aggregation formed by cancer cells has not been available. We examined morphologies of 67 cell lines cultured on three dimensional morphology enhancing NanoCulture Plates (NCP) and classified the types of cellular aggregates that form. Among the 67 cell lines, 49 cell lines formed spheres or spheroids, 8 cell lines formed grape-like aggregation (GLA), 8 cell lines formed other types of aggregation, and 3 cell lines formed monolayer sheets. Seven GLA-forming cell lines were derived from adenocarcinoma among the 8 lines. A neuroendocrine adenocarcinoma cell line PC-3 formed asymmetric GLA with ductal structures on the NCPs and rapidly growing asymmetric tumors that metastasized to lymph nodes in immunocompromised mice. In contrast, another adenocarcinoma cell line DU-145 formed spheroids in vitro and spheroid-like tumors in vivo that did not metastasize to lymph nodes until day 50 after transplantation. Culture in the 3D nanoenvironment and in a defined stem cell medium enabled the neuroendocrine adenocarcinoma cells to form slowly growing large organoids that expressed multiple stem cell markers, neuroendocrine markers, intercellular adhesion molecules, and oncogenes in vitro. In contrast, the more commonly used 2D serum-contained environment reduced intercellular adhesion and induced mesenchymal transition and promoted rapid growth of the cells. In addition, the 3D stemness nanoenvironment promoted secretion of HSP90 and EpCAM-exosomes, a marker of CSC phenotype, from the neuroendocrine organoids. These findings indicate that the NCP-based 3D environment enables cells to form stem cell tumoroids with multipotency and model more accurately the in vivo tumor status at the levels of morphology and gene expression.

    DOI: 10.1371/journal.pone.0191109

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  • OstemiR: a novel panel of microRNA biomarkers in osteoblastic and osteocytic differentiation from mesencymal stem cells. 査読 国際誌

    Takanori Eguchi, Ken Watanabe, Emilio Satoshi Hara, Mitsuaki Ono, Takuo Kuboki, Stuart K Calderwood

    PloS one8 ( 3 ) e58796   2013年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    MicroRNAs (miRNAs) are small RNA molecules of 21-25 nucleotides that regulate cell behavior through inhibition of translation from mRNA to protein, promotion of mRNA degradation and control of gene transcription. In this study, we investigated the miRNA expression signatures of cell cultures undergoing osteoblastic and osteocytic differentiation from mesenchymal stem cells (MSC) using mouse MSC line KUSA-A1 and human MSCs. Ninety types of miRNA were quantified during osteoblastic/osteocytic differentiation in KUSA-A1 cells utilizing miRNA PCR arrays. Coincidently with mRNA induction of the osteoblastic and osteocytic markers, the expression levels of several dozen miRNAs including miR-30 family, let-7 family, miR-21, miR-16, miR-155, miR-322 and Snord85 were changed during the differentiation process. These miRNAs were predicted to recognize osteogenic differentiation-, stemness-, epinegetics-, and cell cycle-related mRNAs, and were thus designated OstemiR. Among those OstemiR, the miR-30 family was classified into miR-30b/c and miR-30a/d/e groups on the basis of expression patterns during osteogenesis as well as mature miRNA structures. In silico prediction and subsequent qRT-PCR in stable miR-30d transfectants clarified that context-dependent targeting of miR-30d on known regulators of bone formation including osteopontin/spp1, lifr, ccn2/ctgf, ccn1/cyr61, runx2, sox9 as well as novel key factors including lin28a, hnrnpa3, hspa5/grp78, eed and pcgf5. In addition, knockdown of human OstemiR miR-541 increased Osteopontin/SPP1 expression and calcification in hMSC osteoblastic differentiation, indicating that miR-541 is a negative regulator of osteoblastic differentiation. These observations indicate stage-specific roles of OstemiR especially miR-541 and the miR-30 family on novel targets in osteogenesis.

    DOI: 10.1371/journal.pone.0058796

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  • Novel transcription-factor-like function of human matrix metalloproteinase 3 regulating the CTGF/CCN2 gene. 査読 国際誌

    Takanori Eguchi, Satoshi Kubota, Kazumi Kawata, Yoshiki Mukudai, Junji Uehara, Toshihiro Ohgawara, Soichiro Ibaragi, Akira Sasaki, Takuo Kuboki, Masaharu Takigawa

    Molecular and cellular biology28 ( 7 ) 2391 - 413   2008年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

    Matrix metalloproteinase 3 (MMP3) is well known as a secretory endopeptidase that degrades extracellular matrices. Recent reports indicated the presence of MMPs in the nucleus (A. J. Kwon et al., FASEB J. 18:690-692, 2004); however, its function has not been well investigated. Here, we report a novel function of human nuclear MMP3 as a trans regulator of connective tissue growth factor (CCN2/CTGF). Initially, we cloned MMP3 cDNA as a DNA-binding factor for the CCN2/CTGF gene. An interaction between MMP3 and transcription enhancer dominant in chondrocytes (TRENDIC) in the CCN2/CTGF promoter was confirmed by a gel shift assay and chromatin immunoprecipitation. The CCN2/CTGF promoter was activated by overexpressed MMP3, whereas a TRENDIC mutant promoter lost the response. Also, the knocking down of MMP3 suppressed CCN2/CTGF expression. By cytochemical and histochemical analyses, MMP3 was detected in the nuclei of chondrocytic cells in culture and also in the nuclei of normal and osteoarthritic chondrocytes in vivo. The nuclear translocation of externally added recombinant MMP3 and six putative nuclear localization signals in MMP3 also were shown. Furthermore, we determined that heterochromatin protein gamma coordinately regulates CCN2/CTGF by interacting with MMP3. The involvement of this novel role of MMP3 in the development, tissue remodeling, and pathology of arthritic diseases through CCN2/CTGF regulation thus is suggested.

    DOI: 10.1128/MCB.01288-07

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  • The Inhibitory Role of Rab11b in Osteoclastogenesis through Triggering Lysosome-Induced Degradation of c-Fms and RANK Surface Receptors. 国際誌

    Manh Tien Tran, Yuka Okusha, Yunxia Feng, Masatoshi Morimatsu, Penggong Wei, Chiharu Sogawa, Takanori Eguchi, Tomoko Kadowaki, Eiko Sakai, Hirohiko Okamura, Keiji Naruse, Takayuki Tsukuba, Kuniaki Okamoto

    International journal of molecular sciences21 ( 24 ) 9352 - 9352   2020年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI AG  

    Rab11b, abundantly enriched in endocytic recycling compartments, is required for the establishment of the machinery of vesicle trafficking. Yet, no report has so far characterized the biological function of Rab11b in osteoclastogenesis. Using in vitro model of osteoclasts differentiated from murine macrophages like RAW-D cells or bone marrow-derived macrophages, we elucidated that Rab11b served as an inhibitory regulator of osteoclast differentiation sequentially via (i) abolishing surface abundance of RANK and c-Fms receptors; and (ii) attenuating nuclear factor of activated T-cells c1 (NFATc-1) upstream signaling cascades, following RANKL stimulation. Rab11b was localized in early and late endosomes, Golgi complex, and endoplasmic reticulum; moreover, its overexpression enlarged early and late endosomes. Upon inhibition of lysosomal function by a specific blocker, chloroquine (CLQ), we comprehensively clarified a novel function of lysosomes on mediating proteolytic degradation of c-Fms and RANK surface receptors, drastically ameliorated by Rab11b overexpression in RAW-D cell-derived osteoclasts. These findings highlight the key role of Rab11b as an inhibitor of osteoclastogenesis by directing the transport of c-Fms and RANK surface receptors to lysosomes for degradation via the axis of early endosomes-late endosomes-lysosomes, thereby contributing towards the systemic equilibrium of the bone resorption phase.

    DOI: 10.3390/ijms21249352

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  • Organoids and Liquid Biopsy in Oral Cancer Research. 国際誌

    Takanori Eguchi

    Journal of clinical medicine9 ( 11 )   2020年11月

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    記述言語:英語  

    To promote the newest discoveries in oral cancer research, a special issue "Frontiers in Oral Cancer-Basic and Clinical Sciences" in the Journal of Clinical Medicine (JCM) was opened from September 2019 to April 2020 [...].

    DOI: 10.3390/jcm9113701

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  • Rab11A Functions as a Negative Regulator of Osteoclastogenesis through Dictating Lysosome-Induced Proteolysis of c-fms and RANK Surface Receptors. 国際誌

    Yuka Okusha, Manh Tien Tran, Mami Itagaki, Chiharu Sogawa, Takanori Eguchi, Tatsuo Okui, Tomoko Kadowaki, Eiko Sakai, Takayuki Tsukuba, Kuniaki Okamoto

    Cells9 ( 11 ) 2384 - 2384   2020年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI AG  

    Osteoclast differentiation and activity are controlled by two essential cytokines, macrophage colony-stimulating factor (M-CSF) and the receptor activator of nuclear factor-κB ligand (RANKL). Rab11A GTPase, belonging to Rab11 subfamily representing the largest branch of Ras superfamily of small GTPases, has been identified as one of the crucial regulators of cell surface receptor recycling. Nevertheless, the regulatory role of Rab11A in osteoclast differentiation has been completely unknown. In this study, we found that Rab11A was strongly upregulated at a late stage of osteoclast differentiation derived from bone marrow-derived macrophages (BMMs) or RAW-D murine osteoclast precursor cells. Rab11A silencing promoted osteoclast formation and significantly increased the surface levels of c-fms and receptor activator of nuclear factor-κB (RANK) while its overexpression attenuated osteoclast formation and the surface levels of c-fms and RANK. Using immunocytochemical staining for tracking Rab11A vesicular localization, we observed that Rab11A was localized in early and late endosomes, but not lysosomes. Intriguingly, Rab11A overexpression caused the enhancement of fluorescent intensity and size-based enlargement of early endosomes. Besides, Rab11A overexpression promoted lysosomal activity via elevating the endogenous levels of a specific lysosomal protein, LAMP1, and two key lysosomal enzymes, cathepsins B and D in osteoclasts. More importantly, inhibition of the lysosomal activity by chloroquine, we found that the endogenous levels of c-fms and RANK proteins were enhanced in osteoclasts. From these observations, we suggest a novel function of Rab11A as a negative regulator of osteoclastogenesis mainly through (i) abolishing the surface abundance of c-fms and RANK receptors, and (ii) upregulating lysosomal activity, subsequently augmenting the degradation of c-fms and RANK receptors, probably via the axis of early endosomes-late endosomes-lysosomes in osteoclasts.

    DOI: 10.3390/cells9112384

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  • Outer membrane vesicles of Porphyromonas gingivalis attenuate insulin sensitivity by delivering gingipains to the liver. 国際誌

    Mariko Seyama, Kaya Yoshida, Kayo Yoshida, Natsumi Fujiwara, Kisho Ono, Takanori Eguchi, Hotaka Kawai, Jiajie Guo, Yao Weng, Yuan Haoze, Kenta Uchibe, Mika Ikegame, Akira Sasaki, Hitoshi Nagatsuka, Kuniaki Okamoto, Hirohiko Okamura, Kazumi Ozaki

    Biochimica et biophysica acta. Molecular basis of disease1866 ( 6 ) 165731 - 165731   2020年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Outer membrane vesicles (OMVs) are nanosized particles derived from the outer membrane of gram-negative bacteria. Oral bacterium Porphyromonas gingivalis (Pg) is known to be a major pathogen of periodontitis that contributes to the progression of periodontal disease by releasing OMVs. The effect of Pg OMVs on systemic diseases is still unknown. To verify whether Pg OMVs affect the progress of diabetes mellitus, we analyzed the cargo proteins of vesicles and evaluated their effect on hepatic glucose metabolism. Here, we show that Pg OMVs were equipped with Pg-derived proteases gingipains and translocated to the liver in mice. In these mice, the hepatic glycogen synthesis in response to insulin was decreased, and thus high blood glucose levels were maintained. Pg OMVs also attenuated the insulin-induced Akt/glycogen synthase kinase-3 β (GSK-3β) signaling in a gingipain-dependent fashion in hepatic HepG2 cells. These results suggest that the delivery of gingipains mediated by Pg OMV elicits changes in glucose metabolisms in the liver and contributes to the progression of diabetes mellitus.

    DOI: 10.1016/j.bbadis.2020.165731

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  • Extracellular Vesicles Enriched with Moonlighting Metalloproteinase Are Highly Transmissive, Pro-Tumorigenic, and Trans-Activates Cellular Communication Network Factor (CCN2/CTGF): CRISPR against Cancer. 国際誌

    Yuka Okusha, Takanori Eguchi, Manh T Tran, Chiharu Sogawa, Kaya Yoshida, Mami Itagaki, Eman A Taha, Kisho Ono, Eriko Aoyama, Hirohiko Okamura, Ken-Ichi Kozaki, Stuart K Calderwood, Masaharu Takigawa, Kuniaki Okamoto

    Cancers12 ( 4 )   2020年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Matrix metalloproteinase 3 (MMP3) plays multiple roles in extracellular proteolysis as well as intracellular transcription, prompting a new definition of moonlighting metalloproteinase (MMP), according to a definition of protein moonlighting (or gene sharing), a phenomenon by which a protein can perform more than one function. Indeed, connective tissue growth factor (CTGF, aka cellular communication network factor 2 (CCN2)) is transcriptionally induced as well as cleaved by MMP3. Moreover, several members of the MMP family have been found within tumor-derived extracellular vesicles (EVs). We here investigated the roles of MMP3-rich EVs in tumor progression, molecular transmission, and gene regulation. EVs derived from a rapidly metastatic cancer cell line (LuM1) were enriched in MMP3 and a C-terminal half fragment of CCN2/CTGF. MMP3-rich, LuM1-derived EVs were disseminated to multiple organs through body fluid and were pro-tumorigenic in an allograft mouse model, which prompted us to define LuM1-EVs as oncosomes in the present study. Oncosome-derived MMP3 was transferred into recipient cell nuclei and thereby trans-activated the CCN2/CTGF promoter, and induced CCN2/CTGF production in vitro. TRENDIC and other cis-elements in the CCN2/CTGF promoter were essential for the oncosomal responsivity. The CRISPR/Cas9-mediated knockout of MMP3 showed significant anti-tumor effects such as the inhibition of migration and invasion of tumor cells, and a reduction in CCN2/CTGF promoter activity and fragmentations in vitro. A high expression level of MMP3 or CCN2/CTGF mRNA was prognostic and unfavorable in particular types of cancers including head and neck, lung, pancreatic, cervical, stomach, and urothelial cancers. These data newly demonstrate that oncogenic EVs-derived MMP is a transmissive trans-activator for the cellular communication network gene and promotes tumorigenesis at distant sites.

    DOI: 10.3390/cancers12040881

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  • Cell Stress Induced Stressome Release Including Damaged Membrane Vesicles and Extracellular HSP90 by Prostate Cancer Cells. 国際誌

    Takanori Eguchi, Chiharu Sogawa, Kisho Ono, Masaki Matsumoto, Manh Tien Tran, Yuka Okusha, Benjamin J Lang, Kuniaki Okamoto, Stuart K Calderwood

    Cells9 ( 3 )   2020年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Tumor cells exhibit therapeutic stress resistance-associated secretory phenotype involving extracellular vesicles (EVs) such as oncosomes and heat shock proteins (HSPs). Such a secretory phenotype occurs in response to cell stress and cancer therapeutics. HSPs are stress-responsive molecular chaperones promoting proper protein folding, while also being released from cells with EVs as well as a soluble form known as alarmins. We have here investigated the secretory phenotype of castration-resistant prostate cancer (CRPC) cells using proteome analysis. We have also examined the roles of the key co-chaperone CDC37 in the release of EV proteins including CD9 and epithelial-to-mesenchymal transition (EMT), a key event in tumor progression. EVs derived from CRPC cells promoted EMT in normal prostate epithelial cells. Some HSP family members and their potential receptor CD91/LRP1 were enriched at high levels in CRPC cell-derived EVs among over 700 other protein types found by mass spectrometry. The small EVs (30-200 nm in size) were released even in a non-heated condition from the prostate cancer cells, whereas the EMT-coupled release of EVs (200-500 nm) and damaged membrane vesicles with associated HSP90α was increased after heat shock stress (HSS). GAPDH and lactate dehydrogenase, a marker of membrane leakage/damage, were also found in conditioned media upon HSS. During this stress response, the intracellular chaperone CDC37 was transcriptionally induced by heat shock factor 1 (HSF1), which activated the CDC37 core promoter, containing an interspecies conserved heat shock element. In contrast, knockdown of CDC37 decreased EMT-coupled release of CD9-containing vesicles. Triple siRNA targeting CDC37, HSP90α, and HSP90β was required for efficient reduction of this chaperone trio and to reduce tumorigenicity of the CRPC cells in vivo. Taken together, we define "stressome" as cellular stress-induced all secretion products, including EVs (200-500 nm), membrane-damaged vesicles and remnants, and extracellular HSP90 and GAPDH. Our data also indicated that CDC37 is crucial for the release of vesicular proteins and tumor progression in prostate cancer.

    DOI: 10.3390/cells9030755

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  • A Novel Model of Cancer Drug Resistance: Oncosomal Release of Cytotoxic and Antibody-Based Drugs. 国際誌

    Takanori Eguchi, Eman Ahmed Taha, Stuart K Calderwood, Kisho Ono

    Biology9 ( 3 )   2020年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Extracellular vesicles (EVs), such as exosomes or oncosomes, often carry oncogenic molecules derived from tumor cells. In addition, accumulating evidence indicates that tumor cells can eject anti-cancer drugs such as chemotherapeutics and targeted drugs within EVs, a novel mechanism of drug resistance. The EV-releasing drug resistance phenotype is often coupled with cellular dedifferentiation and transformation in cells undergoing epithelial-mesenchymal transition (EMT), and the adoption of a cancer stem cell phenotype. The release of EVs is also involved in immunosuppression. Herein, we address different aspects by which EVs modulate the tumor microenvironment to become resistant to anticancer and antibody-based drugs, as well as the concept of the resistance-associated secretory phenotype (RASP).

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  • A reporter system evaluates tumorigenesis, metastasis, β-catenin/MMP regulation, and druggability. 招待 査読 国際誌

    Sogawa C, Eguchi T, C, i, auth, Corresponding author, Okusha Y, Ono K, Ohyama K, Iizuka M, Kawasaki R, Hamada Y, Takigawa M, Sogawa N, Okamoto K, Kozaki K

    Tissue Engineering Part A.25 ( 19-20 ) 1413 - 1425   2019年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Roles of Extracellular HSPs as Biomarkers in Immune Surveillance and Immune Evasion. 国際誌

    Eman A Taha, Kisho Ono, Takanori Eguchi

    International journal of molecular sciences20 ( 18 )   2019年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Extracellular heat shock proteins (ex-HSPs) have been found in exosomes, oncosomes, membrane surfaces, as well as free HSP in cancer and various pathological conditions, also known as alarmins. Such ex-HSPs include HSP90 (α, β, Gp96, Trap1), HSP70, and large and small HSPs. Production of HSPs is coordinately induced by heat shock factor 1 (HSF1) and hypoxia-inducible factor 1 (HIF-1), while matrix metalloproteinase 3 (MMP-3) and heterochromatin protein 1 are novel inducers of HSPs. Oncosomes released by tumor cells are a major aspect of the resistance-associated secretory phenotype (RASP) by which immune evasion can be established. The concepts of RASP are: (i) releases of ex-HSP and HSP-rich oncosomes are essential in RASP, by which molecular co-transfer of HSPs with oncogenic factors to recipient cells can promote cancer progression and resistance against stresses such as hypoxia, radiation, drugs, and immune systems; (ii) RASP of tumor cells can eject anticancer drugs, targeted therapeutics, and immune checkpoint inhibitors with oncosomes; (iii) cytotoxic lipids can be also released from tumor cells as RASP. ex-HSP and membrane-surface HSP (mHSP) play immunostimulatory roles recognized by CD91+ scavenger receptor expressed by endothelial cells-1 (SREC-1)+ Toll-like receptors (TLRs)+ antigen-presenting cells, leading to antigen cross-presentation and T cell cross-priming, as well as by CD94+ natural killer cells, leading to tumor cytolysis. On the other hand, ex-HSP/CD91 signaling in cancer cells promotes cancer progression. HSPs in body fluids are potential biomarkers detectable by liquid biopsies in cancers and tissue-damaged diseases. HSP-based vaccines, inhibitors, and RNAi therapeutics are also reviewed.

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  • Tumor Angiogenic Inhibition Triggered Necrosis (TAITN) in Oral Cancer. 査読 国際誌

    Saori Yoshida, Hotaka Kawai, Takanori Eguchi, Shintaro Sukegawa, May Wathone Oo, Chang Anqi, Kiyofumi Takabatake, Keisuke Nakano, Kuniaki Okamoto, Hitoshi Nagatsuka

    Cells8 ( 7 )   2019年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    CXCR4 is a chemokine receptor crucial in tumor progression, although the angiogenic role of CXCR4 in oral squamous cell carcinoma (OSCC) has not been investigated. Here we show that CXCR4 is crucial for tumor angiogenesis, thereby supporting tumor survival in OSCC. Immunohistochemistry on human clinical specimens revealed that CXCR4 and a tumor vasculature marker CD34 were co-distributed in tumor vessels in human OSCC specimens. To uncover the effects of CXCR4 inhibition, we treated the OSCC-xenografted mice with AMD3100, so-called plerixafor, an antagonist of CXCR4. Notably, we found a unique pathophysiological structure defined as tumor angiogenic inhibition triggered necrosis (TAITN), which was induced by the CXCR4 antagonism. Treatment with AMD3100 increased necrotic areas with the induction of hypoxia-inducible factor-1α in the xenografted tumors, suggesting that AMD3100-induced TAITN was involved in hypoxia and ischemia. Taken together, we demonstrated that CXCR4 plays a crucial role in tumor angiogenesis required for OSCC progression, whereas TAITN induced by CXCR4 antagonism could be an effective anti-angiogenic therapeutic strategy in OSCC treatment.

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  • MZF1 and SCAND1 Reciprocally Regulate CDC37 Gene Expression in Prostate Cancer. 査読 国際誌

    Takanori Eguchi, Thomas L Prince, Manh Tien Tran, Chiharu Sogawa, Benjamin J Lang, Stuart K Calderwood

    Cancers11 ( 6 )   2019年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Cell division control 37 (CDC37) increases the stability of heat shock protein 90 (HSP90) client proteins and is thus essential for numerous intracellular oncogenic signaling pathways, playing a key role in prostate oncogenesis. Notably, elevated expression of CDC37 was found in prostate cancer cells, although the regulatory mechanisms through which CDC37 expression becomes increased are unknown. Here we show both positive and negative regulation of CDC37 gene transcription by two members of the SREZBP-CTfin51-AW1-Number 18 cDNA (SCAN) transcription factor family-MZF1 and SCAND1, respectively. Consensus DNA-binding motifs for myeloid zinc finger 1 (MZF1/ZSCAN6) were abundant in the CDC37 promoter region. MZF1 became bound to these regulatory sites and trans-activated the CDC37 gene whereas MZF1 depletion decreased CDC37 transcription and reduced the tumorigenesis of prostate cancer cells. On the other hand, SCAND1, a zinc fingerless SCAN box protein that potentially inhibits MZF1, accumulated at MZF1-binding sites in the CDC37 gene, negatively regulated the CDC37 gene and inhibited tumorigenesis. SCAND1 was abundantly expressed in normal prostate cells but was reduced in prostate cancer cells, suggesting a potential tumor suppressor role of SCAND1 in prostate cancer. These findings indicate that CDC37, a crucial protein in prostate cancer progression, is regulated reciprocally by MZF1 and SCAND1.

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  • Nicotine promotes lymph node metastasis and cetuximab resistance in head and neck squamous cell carcinoma. 査読 国際誌

    Rieko Shimizu, Soichiro Ibaragi, Takanori Eguchi, Daisuke Kuwajima, Shinichi Kodama, Takashi Nishioka, Tatsuo Okui, Kyoichi Obata, Kiyofumi Takabatake, Hotaka Kawai, Kisho Ono, Kuniaki Okamoto, Hitoshi Nagatsuka, Akira Sasaki

    International journal of oncology54 ( 1 ) 283 - 294   2019年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Epidermal growth factor (EGF) is overexpressed in many cancers and is associated with worse prognosis. EGF binds to its cell surface receptor (EGFR), which induces EGFR phosphorylation. Phosphorylated EGFR (p‑EGFR) is translocated into the nucleus, which increases cancer cell activity. Nicotine, which is one of the main components of tobacco, is absorbed through pulmonary alveoli and mucosal epithelia in the head and neck region by smoking and moves into the blood. Nicotine in blood binds to nicotinic acetylcholine receptor (nAChR) in the central nervous system and serves a crucial role in tobacco addiction. Although nAChR localization is thought to be limited in the nervous system, nAChR is present in a wide variety of non‑neuronal cells, including cancer cells. Recent studies suggest that nicotine contributes to the metastasis and resistance to anti‑cancer drugs of various cancer cells. However, it remains unknown whether head and neck squamous cell carcinoma (HNSCC) cells can utilize nicotine‑nAChR signaling to metastasize and acquire resistance to anti‑cancer drugs, even though the mucosal epithelia of the head and neck region are the primary sites of exposure to tobacco smoke. To the best of our knowledge, the present study is the first to demonstrate the role of nicotine in metastasis and anti‑EGFR‑therapy resistance of HNSCC. The present findings demonstrated that nicotine increased proliferation, migration, invasion, p‑EGFR nuclear translocation and protein kinase B (Akt) phosphorylation in HNSCC cells. It was also demonstrated that nicotine restored cetuximab‑inhibited proliferation, migration and invasion of HNSCC cells. Finally, an in vivo experiment revealed that nicotine increased lymph node metastasis of xenografted tumors, whereas an nAChR inhibitor suppressed lymph node metastasis and p‑EGFR nuclear localization of xenografted tumors. Taken together, these results demonstrated that nicotine induced nuclear accumulation of p‑EGFR, and activation of Akt signaling. These signaling pathways elevated the activities of HNSCC cells, causing lymph node metastasis and serving a role in cetuximab resistance.

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  • Carcinogenic epithelial-mesenchymal transition initiated by oral cancer exosomes is inhibited by anti-EGFR antibody cetuximab. 査読 国際誌

    Toshifumi Fujiwara, Takanori Eguchi, Chiharu Sogawa, Kisho Ono, Jun Murakami, Soichiro Ibaragi, Jun-Ichi Asaumi, Stuart K Calderwood, Kuniaki Okamoto, Ken-Ichi Kozaki

    Oral oncology86   251 - 257   2018年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Overexpression and increased signaling from the epidermal growth factor receptor (EGFR) often changes oral squamous cell carcinoma (OSCC) and thus EGFR is frequently targeted molecularly by the therapeutic antibody cetuximab. We assessed the roles of OSCC-derived extracellular vesicles (EVs), including exosomes in the trafficking of cetuximab and in epithelial-mesenchymal transition (EMT) of epithelial cells. OSCC cells abundantly expressed EGFR, which was secreted from cells with OSCC-EVs upon EGF stimulations. The OSCC-EGFR-EVs were then able to enter into and transform epithelial cells leading to increased mesenchymal traits with increased vimentin and spindle-like shapes. EGF priming of OSCC cells further increased this EMT-initiating effect of the OSCC-EVs. The internalization and pro-EMT effects of the OSCC-EVs were largely blocked by cetuximab. Thus, OSCC-derived EVs transform normal epithelial cells into a mesenchymal phenotype and anti-EGFR therapeutic antibody cetuximab inhibits such a carcinogenic effect of the OSCC-EVs.

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  • Anti-EGFR antibody cetuximab is secreted by oral squamous cell carcinoma and alters EGF-driven mesenchymal transition. 査読 国際誌

    Toshifumi Fujiwara, Takanori Eguchi, Chiharu Sogawa, Kisho Ono, Jun Murakami, Soichiro Ibaragi, Jun-Ichi Asaumi, Kuniaki Okamoto, Stuart K Calderwood, Ken-Ichi Kozaki

    Biochemical and biophysical research communications503 ( 3 ) 1267 - 1272   2018年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Genetic amplification, overexpression, and increased signaling from the epidermal growth factor receptor (EGFR) are often found in oral squamous cell carcinoma (OSCC) and thus EGFR is frequently targeted molecularly by the therapeutic antibody cetuximab. We assessed effects of cetuximab in control of EGF-driven malignant traits of OSCC cells. EGF stimulation promoted progression level of mesenchymal traits in OSCC cells, which were attenuated by cetuximab but incompletely. We pursued a potential mechanism underlying such incomplete attenuation of OSCC malignant traits. Cetuximab promoted secretion of EGFR-EVs by OSCC cells and failed to inhibit EGF-driven secretion of EGFR-EVs. Cetuximab was also found to be robustly secreted with the EGFR-EVs by the OSCC cells. Thus, EGF promotes the level of mesenchymal traits of OSCC cells and secretion of EGFR-EVs, which involve cetuximab resistance.

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  • The intranuclear PEX domain of MMP involves proliferation, migration, and metastasis of aggressive adenocarcinoma cells. 査読 国際誌

    Yuka Okusha, Takanori Eguchi, Chiharu Sogawa, Tatsuo Okui, Keisuke Nakano, Kuniaki Okamoto, Ken-Ichi Kozaki

    Journal of cellular biochemistry119 ( 9 ) 7363 - 7376   2018年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Members of matrix metalloproteinase (MMP) family promote cancer cell migration, invasion, and metastasis through alteration of the tumor milieu, intracellular signaling pathways, and transcription. We examined gene expression signatures of colon adenocarcinoma cell lines with different metastatic potentials and found that rapidly metastatic cells powerfully expressed genes encoding MMP3 and MMP9. The non-proteolytic PEX isoform and proteolytic isoforms of MMPs were significantly expressed in the metastatic cells in vitro. Knockdown of MMP3 attenuated cancer cell migration and invasion in vitro and lung metastasis in vivo. Profound nuclear localization of MMP3/PEX was found in tumor-stroma marginal area. In contrast, MMP9 was localized in central area of subcutaneous tumors. Overexpression of the PEX isoform of MMP3 promoted proliferation and migration of the rapidly metastatic cells in vitro. Taken together, the non-proteolytic PEX isoform of MMPs locating in cell nuclei involves proliferation, migration, and subsequent metastasis of aggressive adenocarcinoma cells.

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  • HSP-enriched properties of extracellular vesicles involve survival of metastatic oral cancer cells. 査読 国際誌

    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 biochemistry119 ( 9 ) 7350 - 7362   2018年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    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.

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  • Genotoxic stress induces Sca-1-expressing metastatic mammary cancer cells. 査読 国際誌

    Jianlin Gong, Benjamin J Lang, Desheng Weng, Takanori Eguchi, Ayesha Murshid, Thiago J Borges, Sachin Doshi, Baizheng Song, Mary A Stevenson, Stuart K Calderwood

    Molecular oncology12 ( 8 ) 1249 - 1263   2018年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We describe a cell damage-induced phenotype in mammary carcinoma cells involving acquisition of enhanced migratory and metastatic properties. Induction of this state by radiation required increased activity of the Ptgs2 gene product cyclooxygenase 2 (Cox2), secretion of its bioactive lipid product prostaglandin E2 (PGE2), and the activity of the PGE2 receptor EP4. Although largely transient, decaying to low levels in a few days to a week, this phenotype was cumulative with damage and levels of cell markers Sca-1 and ALDH1 increased with treatment dose. The Sca-1+ , metastatic phenotype was inhibited by both Cox2 inhibitors and PGE2 receptor antagonists, suggesting novel approaches to radiosensitization.

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  • Depletion of Lipid Efflux Pump ABCG1 Triggers the Intracellular Accumulation of Extracellular Vesicles and Reduces Aggregation and Tumorigenesis of Metastatic Cancer Cells. 査読 国際誌

    Yuri Namba, Chiharu Sogawa, Yuka Okusha, Hotaka Kawai, Mami Itagaki, Kisho Ono, Jun Murakami, Eriko Aoyama, Kazumi Ohyama, Jun-Ichi Asaumi, Masaharu Takigawa, Kuniaki Okamoto, Stuart K Calderwood, Ken-Ichi Kozaki, Takanori Eguchi

    Frontiers in oncology8 ( 376 ) 376 - 376   2018年

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The ATP-binding cassette transporter G1 (ABCG1) is a cholesterol lipid efflux pump whose role in tumor growth has been largely unknown. Our transcriptomics revealed that ABCG1 was powerfully expressed in rapidly metastatic, aggregative colon cancer cells, in all the ABC transporter family members. Coincidently, genetic amplification of ABCG1 is found in 10-35% of clinical samples of metastatic cancer cases. Expression of ABCG1 was further elevated in three-dimensional tumoroids (tumor organoids) within stemness-enhancing tumor milieu, whereas depletion of ABCG1 lowered cellular aggregation and tumoroid growth in vitro as well as hypoxia-inducible factor 1α in cancer cells around the central necrotic areas in tumors in vivo. Notably, depletion of ABCG1 triggered the intracellular accumulation of extracellular vesicles (EVs) and regression of tumoroids. Collectively, these data suggest that ABCG1 plays a crucial role in tumorigenesis in metastatic cancer and that depletion of ABCG1 triggers tumor regression with the accumulation of EVs and their derivatives and cargos, implicating a novel ABCG1-targeting therapeutic strategy by which redundant and toxic substances may be accumulated in tumors leading to their regression.

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  • Monitoring of the Heat Shock Response with a Real-Time Luciferase Reporter. 査読 国際誌

    Toshiki Kijima, Takanori Eguchi, Len Neckers, Thomas L Prince

    Methods in molecular biology (Clifton, N.J.)1709   35 - 45   2018年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Humana Press Inc.  

    The heat shock response (HSR) is a cellular mechanism for counteracting acute proteotoxic stress. In eukaryotes, transcriptional activation of the HSR is regulated by heat shock factor 1 (HSF1). Activation of HSF1 induces the expression of heat shock proteins (HSPs) that function as molecular chaperones to fold and maintain the three-dimensional structure of misfolded proteins. The regulation of the degree and duration of the HSR is controlled by multiple biochemical mechanisms that include posttranslational modification of HSF1 and numerous protein-protein interactions. In this chapter, we describe a method to evaluate the activation and deactivation of the HSR at the transcriptional level using a short half-life luciferase reporter assay. This assay can be used to further characterize the HSR or as a screen for small-molecule inducers, amplifiers, or repressors.

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  • A Tumor Suppressor Gene Product, Platelet-Derived Growth Factor Receptor-Like Protein Controls Chondrocyte Proliferation and Differentiation. 査読 国際誌

    Kazumi Kawata, Satoshi Kubota, Takanori Eguchi, Eriko Aoyama, Norifumi H Moritani, Morihiko Oka, Harumi Kawaki, Masaharu Takigawa

    Journal of cellular biochemistry118 ( 11 ) 4033 - 4044   2017年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    The platelet-derived growth factor receptor-like (PDGFRL) gene is regarded as a tumor suppressor gene. However, nothing is known about the molecular function of PDGFRL. In this study, we initially clarified its function in chondrocytes. Among all cell lines examined, the PDGFRL mRNA level was the highest in chondrocytic HCS-2/8 cells. Interestingly, the proliferation of chondrocytic HCS-2/8 cells was promoted by PDGFRL overexpression, whereas that of the breast cancer-derived MDA-MB-231 cells was inhibited. Of note, in PDGFRL-overexpressing HCS-2/8 cells, the expression of chondrocyte differentiation marker genes, SOX9, ACAN, COL2A1, COL10A1, and ALP, was decreased. Moreover, we confirmed the expression of PDGFRL mRNA in normal cartilage tissue and chondrocytes. Eventually, the expression of PDGFRL mRNA in condrocytes except in the case of hypertrophic chondrocytes was demonstrated in vivo and in vitro. These findings suggest that PDGFRL plays the different roles, depending upon cell types. Particularly, in chondrocytes, PDGFRL may play a new and important role which is distinct from the function previously reported. J. Cell. Biochem. 118: 4033-4044, 2017. © 2017 Wiley Periodicals, Inc.

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  • Catabolic effects of FGF-1 on chondrocytes and its possible role in osteoarthritis. 査読 国際誌

    Abdellatif El-Seoudi, Tarek Abd El Kader, Takashi Nishida, Takanori Eguchi, Eriko Aoyama, Masaharu Takigawa, Satoshi Kubota

    Journal of cell communication and signaling11 ( 3 ) 255 - 263   2017年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    Fibroblast growth factor 1 (FGF-1) is a classical member of the FGF family and is produced by chondrocytes cultured from osteoarthritic patients. Also, this growth factor was shown to bind to CCN family protein 2 (CCN2), which regenerates damaged articular cartilage and counteracts osteoarthritis (OA) in an animal model. However, the pathophysiological role of FGF-1 in cartilage has not been well investigated. In this study, we evaluated the effects of FGF-1 in vitro and its production in vivo by use of an OA model. Treatment of human chondrocytic cells with FGF-1 resulted in marked repression of genes for cartilaginous extracellular matrix components, whereas it strongly induced matrix metalloproteinase 13 (MMP-13), representing its catabolic effects on cartilage. Interestingly, expression of the CCN2 gene was dramatically repressed by FGF-1, which repression eventually caused the reduced production of CCN2 protein from the chondrocytic cells. The results of a reporter gene assay revealed that this repression could be ascribed, at least in part, to transcriptional regulation. In contrast, the gene expression of FGF-1 was enhanced by exogenous FGF-1, indicating a positive feedback system in these cells. Of note, induction of FGF-1 was observed in the articular cartilage of a rat OA model. These results collectively indicate a pathological role of FGF-1 in OA development, which includes an insufficient cartilage regeneration response caused by CCN2 down regulation.

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  • Semaphorin 4D promotes bone invasion in head and neck squamous cell carcinoma. 査読 国際誌

    Hiroyuki Takada, Soichiro Ibaragi, Takanori Eguchi, Tatsuo Okui, Kyoichi Obata, Masanori Masui, Ayaka Morisawa, Kiyofumi Takabatake, Hotaka Kawai, Norie Yoshioka, Nur Mohammad Monsur Hassan, Tsuyoshi Shimo, Guo-Fu Hu, Hitoshi Nagatsuka, Akira Sasaki

    International journal of oncology51 ( 2 ) 625 - 632   2017年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPANDIDOS PUBL LTD  

    Head and neck squamous cell carcinomas (HNSCCs) frequently invade the bones of the facial skeleton. Semaphorin 4D (Sema4D) is an axon guidance molecule produced by oligodendrocytes. Sema4D was also identified in the bone microenvironment and many cancer tissues including HNSCC. To date, however, the role of Sema4D in cancer-associated bone disease is still unknown. This is the first study to demonstrate the role of Sema4D in bone invasion of cancer. In the clinical tissue samples of bone lesion of HNSCC, Sema4D was detected at high levels, and its expression was correlated with insulin-like growth factor-I (IGF-I) expression. In vitro experiments showed that IGF-I regulates Sema4D expression and Sema4D increased proliferation, migration and invasion in HNSCC cells. Sema4D also regulated the expression of receptor activator of nuclear factor κβ ligand (RANKL) in osteoblasts, and this stimulated osteoclastgenesis. Furthermore, knockdown of Sema4D in HNSCC cells inhibited tumor growth and decreased the number of osteoclasts in a mouse xenograft model. Taken together, IGF-I-driven production of Sema4D in HNSCCs promotes osteoclastogenesis and bone invasion.

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  • Intracellular MMP3 Promotes HSP Gene Expression in Collaboration With Chromobox Proteins. 査読 国際誌

    Takanori Eguchi, Stuart K Calderwood, Masaharu Takigawa, Satoshi Kubota, Ken-Ichi Kozaki

    Journal of cellular biochemistry118 ( 1 ) 43 - 51   2017年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Matrix metalloproteinases (MMPs) are crucial factors in tumor progression, inflammatory/immune responses and tissue development/regeneration. Of note, it has been known that MMPs promote genome instability, epithelial-mesenchymal transition, invasion, and metastasis in tumor progression. We previously reported that human MMP3 could translocate into cellular nuclei and control transcription in human chondrosarcoma-derived cells and in articular cartilage (Eguchi et al. [2008] Mol Cell Biol 28(7):2391-2413); however, further transcriptional target genes and cofactors of intranuclear MMP3 have not been uncovered. In this paper, we used transcriptomics analysis in order to examine novel transcriptional target genes regulated by intracellular MMP3. We found that mRNA levels of HSP family members (HSP70B', HSP72, HSP40/DNAJ, and HSP20/CRYAB) are upregulated by the intracellular MMP3 overload. Bioinformatic analysis predicted several transcription factors that possibly interact with MMP3. Among these factors, heat shock factor 1 (HSF1) cooperated with the MMP3 to activate the HSP70B' gene promoter in reporter gene assays, while a dominant negative HSF1 blocked the role for MMP3 in the trans-activation. The hemopexin-like repeat (PEX) domain of the human MMP3 was essential for transcriptional induction of the HSP70B' gene. In addition, chromobox proteins CBX5/HP1α and CBX3/HP1γ cooperated with the PEX domain in induction of HSP70B' mRNA. Taken together, this study newly clarified that intracellular MMP3 cooperate with CBXs/HP1s in transcriptional promotion of HSP genes. J. Cell. Biochem. 118: 43-51, 2017. © 2016 Wiley Periodicals, Inc.

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  • Promoter Analyses of CCN Genes. 査読 国際誌

    Takanori Eguchi, Satoshi Kubota, Masaharu Takigawa

    Methods in molecular biology (Clifton, N.J.)1489   177 - 185   2017年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Humana Press Inc.  

    Promoter analysis is the most basics in the analysis of gene regulation. Luciferase gene is the most commonly used reporter gene in promoter analysis. Luciferase is an enzyme that is used when firefly and Renilla reniformis (sea pansy) emit light. The first experimental step in this reporter gene assay is to connect a particular DNA segment to a luciferase gene. The second step is to transfect the reporter construct into the cells. Thereafter, stable luciferase will be produced with the help of transcriptional machinery, mRNA transporters, and translational machinery in the cells. Luciferase assay measures the quantity of light that is emitted by luciferin-luciferase reaction. Consistent with the fact that CCN2 expression has been shown to be altered by a variety of stimuli, the CCN2 promoter region also haa been shown to be bound and regulated by multiple transcription factors such as Smad, MMP3, NF-κB, AP1, TCF/LEF, and Sox9.

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  • A Novel High-Throughput 3D Screening System for EMT Inhibitors: A Pilot Screening Discovered the EMT Inhibitory Activity of CDK2 Inhibitor SU9516. 査読 国際誌

    Kazuya Arai, Takanori Eguchi, M Mamunur Rahman, Ruriko Sakamoto, Norio Masuda, Tetsuya Nakatsura, Stuart K Calderwood, Ken-Ichi Kozaki, Manabu Itoh

    PloS one11 ( 9 ) e0162394   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    Epithelial-mesenchymal transition (EMT) is a crucial pathological event in cancer, particularly in tumor cell budding and metastasis. Therefore, control of EMT can represent a novel therapeutic strategy in cancer. Here, we introduce an innovative three-dimensional (3D) high-throughput screening (HTS) system that leads to an identification of EMT inhibitors. For the establishment of the novel 3D-HTS system, we chose NanoCulture Plates (NCP) that provided a gel-free micro-patterned scaffold for cells and were independent of other spheroid formation systems using soft-agar. In the NCP-based 3D cell culture system, A549 lung cancer cells migrated, gathered, and then formed multiple spheroids within 7 days. Live cell imaging experiments showed that an established EMT-inducer TGF-β promoted peripheral cells around the core of spheroids to acquire mesenchymal spindle shapes, loss of intercellular adhesion, and migration from the spheroids. Along with such morphological change, EMT-related gene expression signatures were altered, particularly alteration of mRNA levels of ECAD/CDH1, NCAD/CDH2, VIM and ZEB1/TCF8. These EMT-related phenotypic changes were blocked by SB431542, a TGF-βreceptor I (TGFβR1) inhibitor. Inside of the spheroids were highly hypoxic; in contrast, spheroid-derived peripheral migrating cells were normoxic, revealed by visualization and quantification using Hypoxia Probe. Thus, TGF-β-triggered EMT caused spheroid hypoplasia and loss of hypoxia. Spheroid EMT inhibitory (SEMTIN) activity of SB431542 was calculated from fluorescence intensities of the Hypoxia Probe, and then was utilized in a drug screening of EMT-inhibitory small molecule compounds. In a pilot screening, 9 of 1,330 compounds were above the thresholds of the SEMTIN activity and cell viability. Finally, two compounds SB-525334 and SU9516 showed SEMTIN activities in a dose dependent manner. SB-525334 was a known TGFβR1 inhibitor. SU9516 was a cyclin-dependent kinase 2 (CDK2) inhibitor, which we showed also had an EMT-inhibitory activity. The half maximal inhibitory concentration (IC50) of SB-525334 and SU9516 were 0.31 μM and 1.21 μM, respectively, while IC50 of SB431542 was 2.38 μM. Taken together, it was shown that this 3D NCP-based HTS system was useful for screening of EMT-regulatory drugs.

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  • Cellular Reprogramming Using Defined Factors and MicroRNAs. 査読 国際誌

    Takanori Eguchi, Takuo Kuboki

    Stem cells international2016   7530942 - 7530942   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Hindawi Publishing Corporation  

    Development of human bodies, organs, and tissues contains numerous steps of cellular differentiation including an initial zygote, embryonic stem (ES) cells, three germ layers, and multiple expertized lineages of cells. Induced pluripotent stem (iPS) cells have been recently developed using defined reprogramming factors such as Nanog, Klf5, Oct3/4 (Pou5f1), Sox2, and Myc. This outstanding innovation is largely changing life science and medicine. Methods of direct reprogramming of cells into myocytes, neurons, chondrocytes, and osteoblasts have been further developed using modified combination of factors such as N-myc, L-myc, Sox9, and microRNAs in defined cell/tissue culture conditions. Mesenchymal stem cells (MSCs) and dental pulp stem cells (DPSCs) are also emerging multipotent stem cells with particular microRNA expression signatures. It was shown that miRNA-720 had a role in cellular reprogramming through targeting the pluripotency factor Nanog and induction of DNA methyltransferases (DNMTs). This review reports histories, topics, and idea of cellular reprogramming.

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  • Targeting the hsp70 gene delays mammary tumor initiation and inhibits tumor cell metastasis 査読

    J. Gong, D. Weng, T. Eguchi, A. Murshid, M. Y. Sherman, B. Song, S. K. Calderwood

    ONCOGENE34 ( 43 ) 5460 - 5471   2015年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Elevated levels of the inducible heat-shock protein 70 (Hsp72) have been implicated in mammary tumorigenesis in histological investigations of human breast cancer. We therefore examined the role of Hsp72 in mice, using animals in which the hsp70 gene was inactivated. We used a spontaneous tumor system with mice expressing the polyomavirus middle T (PyMT) oncogene under control of the mouse mammary tumor virus (MMTV) long-terminal repeat (MMT mice). These mice developed spontaneous, metastatic mammary cancer. We then showed Hsp72 to be upregulated in a fraction of mammary cancer initiating cells (CIC) within the MMT tumor cell population. These cells were characterized by elevated surface levels of stem cell markers CD44 and Sca1 and by rapid metastasis. Inactivation of the hsp70 gene delayed the initiation of mammary tumors. This delay in tumor initiation imposed by loss of hsp70 was correlated with a decreased pool of CIC. Interestingly, hsp70 knockout significantly reduced invasion and metastasis by mammary tumor cells and implicated its product Hsp72 in cell migration and formation of secondary neoplasms. Impaired tumorigenesis and metastasis in hsp70-knockout MMT mice was associated with downregulation of the met gene and reduced activition of the oncogenic c-Met protein. These experiments therefore showed Hsp72 to be involved in the growth and progression of mammary carcinoma and highlighted this protein as a potential target for anticancer drug development.

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  • Role and Regulation of Myeloid Zinc Finger Protein 1 in Cancer. 査読 国際誌

    Taka Eguchi, Thomas Prince, Barbara Wegiel, Stuart K Calderwood

    Journal of cellular biochemistry116 ( 10 ) 2146 - 54   2015年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Myeloid zinc finger 1 (MZF1) belongs to the SCAN-Zinc Finger (SCAN-ZF) transcription factor family that has recently been implicated in a number of types of cancer. Although the initial studies concentrated on the role of MZF1 in myeloid differentiation and leukemia, the factor now appears to be involved in the etiology of major solid tumors such as lung, cervical, breast, and colorectal cancer. Here we discuss the regulation of MZF1 that mediated its recruitment and activation in cancer, concentrating on posttranslational modification by phosphorylation, and sumoylation, formation of promyelocytic leukemia nuclear bodies and its association with co-activators and co-repressors.

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  • HSF1 regulation of β-catenin in mammary cancer cells through control of HuR/elavL1 expression. 査読 国際誌

    Chou SD, Murshid A, Eguchi T, Gong J, Calderwood SK

    Oncogene34 ( 17 ) 2178 - 2188   2015年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • OstemiR: A Novel Panel of MicroRNA Biomarkers in Osteoblastic and Osteocytic Differentiation from Mesencymal Stem Cells 査読

    Eguchi, Takanori, Hara, Emilio Satoshi, Ono, Mitsuaki, Watanabe, Ken, Kuboki, Takuo, Calderwood, Stuart

    Faseb Journal29 ( 3 ) e58796   2015年

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    掲載種別:研究論文(学術雑誌)  

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  • miRNA-720 regulates the stem cell phenotype and differentiation of human dental pulp-derived mesenchymal stromal cells 査読

    Hara Emilio Satoshi, Ono Mitsuaki, Eguchi Takanori, Hai Thanh Pham, Tajima Shoji, Calderwood Stuart K, Matsumoto Takuya, Kuboki Takuo, IEEE

    2014 International Symposium on Micro-Nanomechatronics and Human Science (Mhs)   2014年

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    掲載種別:研究論文(学術雑誌)  

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  • Stress proteins in aging and life span. 査読 国際誌

    Ayesha Murshid, Takanori Eguchi, Stuart K Calderwood

    International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group29 ( 5 ) 442 - 7   2013年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INFORMA HEALTHCARE  

    Heat shock proteins (HSP) are molecular chaperones and have been implicated in longevity and aging in many species. Their major functions include chaperoning misfolded or newly synthesised polypeptides, protecting cells from proteotoxic stress, and processing of immunogenic agents. These proteins are expressed constitutively and can be induced by stresses such as heat, oxidative stress and many more. The induction of HSP in aging could potentially maintain protein homeostasis and longevity by refolding the damaged proteins which accumulate during aging and are toxic to cells. HSP are shown to increase life span in model organisms such as Caenorhabditis elegans and decrease aging-related proteotoxicity. Thus, decrease in HSP in aging is associated with disruption of cellular homeostasis which causes diseases such as cancer, cell senescence and neurodegeneration. HSP levels are decreased with aging in most organs including neurons. Aging also causes attenuation or alteration of many signalling pathways as well as the expression of transcription factors such as heat shock factor (HSF). The alteration in regulation and synthesis of Forkhead box O3a (FoxO3a) family of transcription factors as well as major antioxidant enzymes (manganese superoxide dismutase, catalase) are also seen in aging. Among many signalling mechanisms involved in altering longevity and aging, the insulin/IGF-1 pathway and the Sir2 deacetylase are highly significant. This review enquires into the role of some of these pathways in longevity/aging along with HSP.

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  • miRNA-720 controls stem cell phenotype, proliferation and differentiation of human dental pulp cells. 査読 国際誌

    Emilio Satoshi Hara, Mitsuaki Ono, Takanori Eguchi, Satoshi Kubota, Hai Thanh Pham, Wataru Sonoyama, Shoji Tajima, Masaharu Takigawa, Stuart K Calderwood, Takuo Kuboki

    PloS one8 ( 12 ) e83545   2013年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Dental pulp cells (DPCs) are known to be enriched in stem/progenitor cells but not well characterized yet. Small non-coding microRNAs (miRNAs) have been identified to control protein translation, mRNA stability and transcription, and have been reported to play important roles in stem cell biology, related to cell reprogramming, maintenance of stemness and regulation of cell differentiation. In order to characterize dental pulp stem/progenitor cells and its mechanism of differentiation, we herein sorted stem-cell-enriched side population (SP) cells from human DPCs and periodontal ligament cells (PDLCs), and performed a locked nucleic acid (LNA)-based miRNA array. As a result, miR-720 was highly expressed in the differentiated main population (MP) cells compared to that in SP cells. In silico analysis and a reporter assay showed that miR-720 targets the stem cell marker NANOG, indicating that miR-720 could promote differentiation of dental pulp stem/progenitor cells by repressing NANOG. Indeed, gain-and loss-of-function analyses showed that miR-720 controls NANOG transcript and protein levels. Moreover, transfection of miR-720 significantly decreased the number of cells positive for the early stem cell marker SSEA-4. Concomitantly, mRNA levels of DNA methyltransferases (DNMTs), which are known to play crucial factors during stem cell differentiation, were also increased by miR-720 through unknown mechanism. Finally, miR-720 decreased DPC proliferation as determined by immunocytochemical analysis against ki-67, and promoted odontogenic differentiation as demonstrated by alizarin red staining, as well as alkaline phosphatase and osteopontin mRNA levels. Our findings identify miR-720 as a novel miRNA regulating the differentiation of DPCs.

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  • Role of LRP1 in transport of CCN2 protein in chondrocytes. 査読 国際誌

    Kazumi Kawata, Satoshi Kubota, Takanori Eguchi, Eriko Aoyama, Norifumi H Moritani, Seiji Kondo, Takashi Nishida, Masaharu Takigawa

    Journal of cell science125 ( Pt 12 ) 2965 - 72   2012年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Low-density lipoprotein receptor-related protein 1 (LRP1) is known to be a receptor for signal transmission and endocytosis. We have previously reported that LRP1 regulates WNT-β-catenin and protein kinase C signaling in chondrocytes, represses the hypertrophy of chondrocytes during endochondral ossification and that LRP1 is colocalized with a ligand, CCN family member 2 (CCN2; also known as connective tissue growth factor, CTGF), which conducts endochondral ossification, in chondrocytes. However, the role of LRP1 in the endocytic transport of CCN2 in chondrocytes is not yet understood. In the present study, we investigated the interaction between LRP1 and CCN2 during endocytic trafficking. Small interfering RNA (siRNA)-mediated knockdown of LRP1 in chondrocytic HCS-2/8 cells showed that the amount of exogenous CCN2 binding and/or incorporation was decreased in the LRP1 downregulated cells. Importantly, we observed that CCN2 internalization in chondrocytes was dependent on clathrin, and internalizated CCN2 was colocalized with an early or recycling endosome marker. Transcytosis of CCN2 through HCS-2/8 cells was confirmed by performing experiments with a trans-well apparatus, and the amount of transcytosed CCN2 was decreased by an LRP1 antagonist. These findings rule out possible leakage and confirm the crucial involvement of LRP1 during experimental transcytosis. Moreover, under hypoxic conditions that mimic the cartilaginous microenvironment, the level of LRP1 and the amount of transcytosed CCN2 increased, and these increases were neutralized by treatment with the LRP1 antagonist. The distribution of LRP1 and its antagonist in the growth plate in vivo was consistent with that of CCN2 in this tissue, which is produced by and transported by LRP1 from the chondrocytes in the prehypertrophic layer. These findings suggest that LRP1 mediates the transcytosis of CCN2, which might be a crucial event that determines the distribution of CCN2 in cartilage.

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  • Role of low-density lipoprotein receptor related protein 1 (LRP1) in CCN2/connective tissue growth factor (CTGF) protein transport in chondrocytes

    Kawata., K, Kubota, S, Eguchi, T, Aoyama, E, Moritani, N, Kondo, S, Nishida, T, Takigawa, M

    J Cell Sci15   2965 - 2972   2012年

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  • A coding RNA segment that enhances the ribosomal recruitment of chicken ccn1 mRNA. 査読 国際誌

    Yoshiki Mukudai, Satoshi Kubota, Takanori Eguchi, Kumi Sumiyoshi, Danilo Janune, Seiji Kondo, Satoru Shintani, Masaharu Takigawa

    Journal of cellular biochemistry111 ( 6 ) 1607 - 18   2010年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    CCN1, a member of the CCN family of proteins, plays important physiological or pathological roles in a variety of tissues. In the present study, we initially found a highly guanine-cytosine (GC)-rich region of approximately 200 bp near the 5'-end of the open reading frame, which was always truncated by amplification of the corresponding cDNA region through the conventional polymerase chain reaction. An RNA in vitro folding assay and selective ribonuclease digestion of the corresponding segment of the ccn1 mRNA confirmed the involvement of a stable secondary structure. Subsequent RNA electromobility-shift assays demonstrated the specific binding of some cytoplasmic factor(s) in chicken embryo fibroblasts to the RNA segment. Moreover, the corresponding cDNA fragment strongly enhanced the expression of the reporter gene in cis at the 5'-end, but did not do so at the 3'-end. According to the results of a ribosomal assembly test, the effect of the mRNA segment can predominantly be ascribed to the enhancement of transport and/or entry of the mRNA into the ribosome. Finally, the minimal GC-rich mRNA segment that was predicted and demonstrated to form a secondary structure was confirmed to be a functional regulatory element. Thus, we here uncover a novel dual-functionality of the mRNA segment in the ccn1 open reading frame, which segment acts as a cis-element that mediates posttranscriptional gene regulation, while retaining the information for the amino acid sequence of the resultant protein.

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  • Role of the low-density lipoprotein receptor-related protein-1 in regulation of chondrocyte differentiation. 査読 国際誌

    Kazumi Kawata, Satoshi Kubota, Takanori Eguchi, Norifumi H Moritani, Tsuyoshi Shimo, Seiji Kondo, Takashi Nishida, Shogo Minagi, Masaharu Takigawa

    Journal of cellular physiology222 ( 1 ) 138 - 48   2010年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The low-density lipoprotein receptor-related protein 1 (LRP1) is known as an endocytic and signal transmission receptor. We formerly reported the gene expression and the localization of LRP1 in cartilage tissue and chondrocytes, but its roles in the differentiation of chondrocytes remained to be investigated. Here, in order to address this issue, we employed RNAi strategy to knockdown lrp1 in chondrocytic cells and obtained findings indicating a critical role therein. As a result of lrp1 knockdown, aggrecan and col2a1 mRNA levels were decreased. However, that of col10a1 or mmp13 mRNA was rather increased. Under this condition, we performed a promoter assay for Axin2, which is known to be induced by activation of the WNT/beta-catenin (betacat) signaling pathway. Thereby, we found that Axin2 promoter activity was enhanced in the lrp1 knockdown cells. Furthermore, when the WNT/beta-catenin pathway was activated in chondrocytic cells by WNT3a or SB216763, which inhibits the phosphorylation of GSK3beta, the mRNA levels of aggrecan and col2a1 were decreased, whereas that of mmp13 was increased. Additionally, the level of phosphorylated protein kinase C (PKC) zeta was also decreased in the lrp1 knockdown cells. When the phosphorylation of PKCzeta was selectively inhibited, aggrecan and col2a1 mRNA levels decreased, whereas the mmp13 mRNA level increased. These data demonstrate that LRP1 exerts remarkable effects to retain the mature phenotype of chondrocytes as a critical mediator of cell signaling. Our findings also indicate that the onset of hypertrophy during endochondral ossification appears to be particularly dependent on the WNT and PKC signaling initiated by LRP1.

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  • Novel Transcriptional Regulation of CCN2/CTGF by Nuclear Translocation of MMP3 査読

    Takanori Eguchi, Satoshi Kubota, Kazumi Kawata, Yoshiki Mukudai, Junji Uehara, Toshihiro Ohgawara, Soichiro Ibaragi, Akira Sasaki, Takuo Kuboki, Masaharu Takigawa

    CCN PROTEINS IN HEALTH AND DISEASE: AN OVERVIEW OF THE FIFTH INTERNATIONAL WORKSHOP ON THE CCN FAMILY OF GENES   255 - +   2010年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:SPRINGER-VERLAG BERLIN  

    CCN2/CTGF, previously known as Connective Tissue Growth Factor, is a crucial regulator of extra-cellular matrix (ECM), which promotes ECM synthesis and stabilization. As their family name clearly implies, matrix metalloproteases (MMPs) are also localized in the ECM, where they function as proteases, modulating cell signaling by cleaving proteins such as matrix proteins, growth factors and growth factor receptors. Strong expression of CCN2/CTGF in chondrocytic cells occurs through transcription enhancer dominant in chondrocytes (TRENDIC). Matrix metalloprotease-3 (MMP3) is a novel TRENDIC-binding transcription factor for CCN2/CTGF expression. First, MMP3 cDNA was cloned as a TRENDIC-binding factor by Southwestern screening. The interaction between MMP3 and TRENDIC was confirmed by a gel shift assay and chromatin immunoprecipitation. The CCN2/CTGF promoter was activated by transfected MMP3, whereas a TRENDIC mutant for the promoter lost the response. In addition, the knockdown of MMP3 suppressed CCN2/CTGF expression. Cytochemical and histochemical analyses demonstrated that MMP3 was detected in the nuclei of chondrocytic cells in culture and also in the nuclei of normal and osteoarthritic chondrocytes in vivo. The nuclear translocation of externally added recombinant MMP3 was observed in 30 min after the addition, and six putative nuclear localization signals were found in MMP3. These results indicated a novel trans-activation mechanism of CCN2/CTGF by the nuclear translocation of MMP3 through binding with TRENDIC in chondrocytes. Although MMPs historically had been recognized as a protease for extra-cellular proteins, this study indicated that it also stimulates ECM synthesis through CCN2/CTGF trans-activation. This novel regulatory role of the ECM may contribute to understanding the mechanism of not only the development, but also the pathogenesis of arthritis fibrosis and periodontitis.

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  • Nucleophosmin/B23: A Multifunctional Regulator that Determines the Fate of CCN2 mRNA 査読

    Satoshi Kubota, Yoshiki Mukudai, Harumi Kawaki, Seiji Kondo, Takanori Eguchi, Kumi Sumiyoshi, Toshihiro Ohgawara, Tsuyoshi Shimo, Masaharu Takigawa

    CCN PROTEINS IN HEALTH AND DISEASE: AN OVERVIEW OF THE FIFTH INTERNATIONAL WORKSHOP ON THE CCN FAMILY OF GENES   41 - +   2010年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:SPRINGER-VERLAG BERLIN  

    CCN2/CTGF is a multifunctional molecule that has been shown to play a central role in chondrocyte differentiation. During this process, the expression of ccn2 is tightly regulated to confer a maximal level at prehypertrophic - hypertrophic stages, in which the 3'-untranslated region (UTR) of the mRNA is critically involved in mediating its post-transcriptional regulation. In our previous studies, we found that a 40-kDa protein binding specifically to an RNA cis-element, 3'-100/50, in the 3'-UTR of the chicken ccn2 mRNA regulated the intracellular stability of the mRNA. The interaction of this 40-kDa protein with 3'-100/50 was enhanced in proliferating chondrocytes, in which ccn2 mRNA is rapidly degraded; whereas a prolonged half life of ccn2 mRNA is observed in hypertrophic chondrocytes, where the interaction of the 40 kDa-protein and 3'-100/50 is diminished. Collectively, the data suggested that this 40-kDa protein acts as a ccn2-specific mRNA destabilizer during chondrocyte differentiation.
    In this present study we finally identified this 40-kDa protein as nucleophosmin (NPM)/B23. NPM is a nuclear-cytoplasmic shuttling protein that is characterized by its multiple functionality. This protein is known to be a histone chaperone, a regulator of ribosomal RNA transcription, as well as an RNA-binding post-transcriptional regulator of gene expression. In our hands, direct binding of NPM to 3'-100/50 was confirmed not only by RNA EMSA and UV crosslinking assays, but also by RNA immunoprecipitation analysis. By using recombinant chicken NPM, we could successfully reconstitute the post-transcriptional regulation of ccn2 by NPM in vitro and found that this regulation was more robust in chondrocytes than in fibroblasts. Furthermore, siRNA-mediated gene silencing of NPM in vivo clearly showed enhanced ccn2 gene expression and a prolonged half life of the ccn2 mRNA, confirming the functional property of NPM as a specific destabilizer of the ccn2 mRNA in living cells.
    The 5'-100/50 element, a target of NPM, is evolutionally conserved among vertebrate species. Therefore, we consider NPM to be a critical post-transcriptional regulator of ccn2 acting via 3'-UTR during endochondral ossification and possibly, in other physiological and pathological states as well.

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  • Regulation of chondrocytic phenotype by micro RNA 18a: involvement of Ccn2/Ctgf as a major target gene. 査読 国際誌

    Toshihiro Ohgawara, Satoshi Kubota, Harumi Kawaki, Seiji Kondo, Takanori Eguchi, Naito Kurio, Eriko Aoyama, Akira Sasaki, Masaharu Takigawa

    FEBS letters583 ( 6 ) 1006 - 10   2009年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    We searched for miRNAs that were down-regulated in chondrocytic cells and predicted to target CCN2/connective tissue growth factor (CCN2/CTGF) that promotes endochondral ossification. Among them, expression of miR-18a was most strongly repressed in chondrocytic cells. Reporter gene analysis confirmed the functionality of an miR-18a target in the 3'-untranslated region of Ccn2 mRNA, which was predicted in silico. Indeed, introduction of miR-18a efficiently repressed the CCN2 production from chondrocytic cells. Finally, transfected miR-18a significantly repressed the mature chondrocytic phenotype. Our present study revealed a regulatory role for miR-18a in chondrocytic differentiation through CCN2.

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  • Posttranscriptional regulation of chicken ccn2 gene expression by nucleophosmin/B23 during chondrocyte differentiation. 査読 国際誌

    Yoshiki Mukudai, Satoshi Kubota, Harumi Kawaki, Seiji Kondo, Takanori Eguchi, Kumi Sumiyoshi, Toshihiro Ohgawara, Tsuyoshi Shimo, Masaharu Takigawa

    Molecular and cellular biology28 ( 19 ) 6134 - 47   2008年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

    CCN2/CTGF is a multifunctional factor that plays a crucial role in the growth and differentiation of chondrocytes. The chicken ccn2 gene is regulated not only at the transcriptional level but also by the interaction between a posttranscriptional element in the 3' untranslated region (3'-UTR) and a cofactor. In the present study, we identified a nucleophosmin (NPM) (also called B23) as this cofactor. Binding of NPM to the element was confirmed, and subsequent analysis revealed a significant correlation between the decrease in cytosolic NPM and the increased stability of the ccn2 mRNA during chondrocyte differentiation in vivo. Furthermore, recombinant chicken NPM enhanced the degradation of chimeric RNAs containing the posttranscriptional cis elements in a chicken embryonic fibroblast extract in vitro. It is noteworthy that the RNA destabilization effect by NPM was far more prominent in the cytosolic extract of chondrocytes than in that of fibroblasts, representing a chondrocyte-specific action of NPM. Stimulation by growth factors to promote differentiation changed the subcellular distribution of NPM in chondrocytes, which followed the expected patterns from the resultant change in the ccn2 mRNA stability. Therefore, the present study reveals a novel aspect of NPM as a key player in the posttranscriptional regulation of ccn2 mRNA during the differentiation of chondrocytes.

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  • Gene expression and distribution of connective tissue growth factor (CCN2/CTGF) during secondary ossification center formation. 査読 国際誌

    Morihiko Oka, Satoshi Kubota, Seiji Kondo, Takanori Eguchi, Chisa Kuroda, Kazumi Kawata, Shogo Minagi, Masaharu Takigawa

    The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society55 ( 12 ) 1245 - 55   2007年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:HISTOCHEMICAL SOC INC  

    CCN2/connective tissue growth factor (CCN2/CTGF) is a critical signaling modulator of mesenchymal tissue development. This study investigated the localization and expression of CCN2/CTGF as a factor supporting angiogenesis and chondrogenesis during development of secondary ossification centers in the mouse tibial epiphysis. Formation of the secondary ossification center was initiated by cartilage canal formation and blood vessel invasion at 7 days of age, and onset of ossification was observed at 14 days. In situ hybridization showed that CCN2/CTGF mRNA was distinctively expressed in the region of the cartilage canal and capsule-attached marginal tissues at 7 days of age, and distinct expression was also observed in proliferating chondrocytes around the marrow space at 14 days of age. Immunostaining showed that CCN2/CTGF was distributed broadly around the expressed cells located in the central region of the epiphysis, where the chondrocytes become hypertrophic and the cartilage canal enters into the hypertrophic mass. Furthermore, an overlapping distribution of metalloproteinase (MMP)9 and CCN2/CTGF was found in the secondary ossification center. These findings suggest that the CCN2/CTGF is involved in establishing epiphyseal vascularization and remodeling, which eventually determines the secondary ossification center in the developing epiphysial cartilage.

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  • Different transcriptional strategies for ccn2/ctgf gene induction between human chondrocytic and breast cancer cell lines. 査読 国際誌

    Takanori Eguchi, Satoshi Kubota, Kazumi Kawata, Yoshiki Mukudai, Toshihiro Ohgawara, Kohei Miyazono, Kyouji Nakao, Seiji Kondo, Masaharu Takigawa

    Biochimie89 ( 3 ) 278 - 88   2007年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER  

    Connective tissue growth factor (CTGF/CCN2) plays a critical role in endochondral bone formation; however, CCN2 also promotes angiogenesis and bone metastasis in breast cancer. Chondrocytic HCS-2/8 cells and breast cancer MDA231 cells produce over 6 times more CCN2 than any other cell type. In this study, we demonstrate that these cell lines employ different transcriptional strategies for ccn2 gene induction. Four tandem copies of the dominant transcriptional enhancer in chondrocytes (4 x TRENDIC) were chimerically connected to an SV40 promoter-luciferase construct and subsequently analyzed. The enhancement of the promoter activity by 4 x TRENDIC was greater in the HCS-2/8 cells (7-fold) than in the other 4 cell lines (3-4 fold). The TRENDIC-binding protein complex was detected at a higher signal in the HCS-2/8 cells than in the other cell lines. In addition, the HCS-2/8 nuclear factors strongly targeted not only TRENDIC, but also the previously reported basal control element and a novel enhancer element in the ccn2 promoter. In contrast, high-level ccn2 gene induction in MDA231 cells was largely dependent on Smad signaling through the Smad-binding element in the ccn2 promoter. Based on these results, we propose a model of differential transcription of the ccn2 gene between the chondrocytic cell line and the breast cancer cell line, and therefore imply that these cells utilize distinct transcriptional strategies to obtain the enhanced CCN2 production that is not observed in other types of cells.

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  • Pathogenic role of connective tissue growth factor (CTGF/CCN2) in osteolytic metastasis of breast cancer. 査読 国際誌

    Tsuyoshi Shimo, Satoshi Kubota, Norie Yoshioka, Soichiro Ibaragi, Sachiko Isowa, Takanori Eguchi, Akira Sasaki, Masaharu Takigawa

    Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research21 ( 7 ) 1045 - 59   2006年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BONE & MINERAL RES  

    UNLABELLED: The role of CTGF/CCN2 in osteolytic metastasis by breast cancer cells and its mechanism of action were studied. Osteolytic metastasis accompanied by CCN2 and PTHrP overproduction was efficiently inhibited by an anti-CCN2 antibody. Furthermore, we found that CCN2 was induced by PTHrP through PKA-, PKC-, and ERK-mediated pathways therein. INTRODUCTION: Connective tissue growth factor (CTGF/CCN2) is a mediator of local angiogenesis induced by breast cancer, but its role in osteolytic metastasis has not been evaluated. PTH-related peptide (PTHrP) is another critical factor in the development of the osteolytic metastasis. Using both in vivo and in vitro approaches, we studied whether/how neutralization of CCN2 prevented bone metastasis and how PTHrP signaling is related. MATERIALS AND METHODS: A mouse model of bone metastasis by human breast cancer cell line MDA231 was treated with a CCN2-neutralizing antibody, and osteolytic bone metastases were assessed on radiographs and immunohistochemistry. Ccn2 gene expression and transcription were examined by Northern blot and luciferase analysis. Immunoblot analysis and kinase inhibitors were used to identify the signaling pathways implicated. Anti-angiogenic/osteoclastogenic effects of ccn2 downregulation were also evaluated. RESULTS: Treatment of mice with a CCN2-neutralizing antibody greatly decreased osteolytic bone metastasis, microvasculature, and osteoclasts involved. The antibody also suppressed the growth of subcutaneous tumor in vivo and proliferation and migration of human umbilical vein endothelial cells (HUVECs) in vitro. Downregulation of ccn2 also repressed osteoclastogenesis. CCN2 expression was specifically observed in cancer cells producing PTHrP and type I PTH/PTHrP receptor (PTH1R) invaded the bone marrow, and PTHrP strongly upregulated ccn2 in MDA231 cells in vitro. Activation of protein kinase C (PKC) and protein kinase A (PKA) was necessary and sufficient for the stimulation of ccn2 by PTHrP. Indeed, inhibition of the extracellular signal-regulated kinase (ERK1/2), PKC, or PKA by specific inhibitors counteracted the stimulation of ccn2 expression. Incubation of MDA231 cells with PTHrP induced the activation of ERK1/2. Consistent with these findings, inhibition of PKC prevented PTHrP-induced ERK1/2 activation, whereas 12-O-tetradecanoylphorbol13-acetate (TPA), a stimulator of PKC, upregulated it. CONCLUSIONS: CCN2 was critically involved in osteolytic metastasis and was induced by PKA- and PKC-dependent activation of ERK1/2 signaling by PTHrP. Thus, CCN2 may be a new molecular target for anti-osteolytic therapy to shut off the PTHrP-CCN2 signaling pathway.

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  • Possible role of LRP1, a CCN2 receptor, in chondrocytes. 査読 国際誌

    Kazumi Kawata, Takanori Eguchi, Satoshi Kubota, Harumi Kawaki, Morihiko Oka, Shogo Minagi, Masaharu Takigawa

    Biochemical and biophysical research communications345 ( 2 ) 552 - 9   2006年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Low density lipoprotein receptor (LDLR)-related protein 1 (LRP1/CD91) is one of the receptors of CCN2 that conducts endochondral ossification and cartilage repair. LRP1 is a well-known endocytic receptor, but its distribution among chondrocytes remains to be elucidated. We herein demonstrate for the first time that the distribution of LRP1 in chondrocytes except for hypertrophic chondrocytes in vivo and in vitro. Interestingly, the LRP1 levels were higher in mature chondrocytic HCS-2/8 and osteoblastic SaOS-2 than in other cells, whereas the other LDLR family members involved in ossification were detected at lower levels in HCS-2/8. It was interesting to note that in HCS-2/8, LRP1 was observed not only on the cell surface and in the cytoplasm, but also in the nucleus. Exogenously added CCN2 was incorporated into HCS-2/8, which was partially co-localized with LRP1, and targeted to the recycling endosomes and nucleus as well as the lysosomes. These findings suggest specific roles of LRP1 in cartilage biology.

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  • Collaborative action of M-CSF and CTGF/CCN2 in articular chondrocytes: possible regenerative roles in articular cartilage metabolism. 査読 国際誌

    Kyouji Nakao, Satoshi Kubota, Hideyuki Doi, Takanori Eguchi, Morihiko Oka, Takuo Fujisawa, Takashi Nishida, Masaharu Takigawa

    Bone36 ( 5 ) 884 - 92   2005年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE INC  

    It is known that expression of the macrophage colony-stimulating factor (M-CSF) gene is induced in articular chondrocytes upon inflammation. However, the functional role of M-CSF in cartilage has been unclear. In this study, we describe possible roles of M-CSF in the protection and maintenance of the articular cartilage based on the results of experiments using human chondrocytic cells and rat primary chondrocytes. Connective tissue growth factor (CTGF/CCN2) is known to be a potent molecule to regenerate damaged cartilage by promoting the growth and differentiation of articular chondrocytes. Here, we uncovered the fact that M-CSF induced the mRNA expression of the ctgf/ccn2 gene in those cells. Enhanced production of CTGF/CCN2 protein by M-CSF was also confirmed. Furthermore, M-CSF could autoactivate the m-csf gene, forming a positive feed-back network to amplify and prolong the observed effects. Finally, promotion of proteoglycan synthesis was observed by the addition of M-CSF. These findings taken together indicate novel roles of M-CSF in articular cartilage metabolism in collaboration with CTGF/CCN2, particularly during an inflammatory response. Such roles of M-CSF were further supported by the distribution of M-CSF producing chondrocytes in experimentally induced rat osteoarthritis cartilage in vivo.

    DOI: 10.1016/j.bone.2004.10.015

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  • Regulation of chicken ccn2 gene by interaction between RNA cis-element and putative trans-factor during differentiation of chondrocytes. 査読 国際誌

    Yoshiki Mukudai, Satoshi Kubota, Takanori Eguchi, Seiji Kondo, Kyouji Nakao, Masaharu Takigawa

    The Journal of biological chemistry280 ( 5 ) 3166 - 77   2005年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    CCN2/CTGF is a multifunctional growth factor. Our previous studies have revealed that CCN2 plays important roles in both growth and differentiation of chondrocytes and that the 3'-untranslated region (3'-UTR) of ccn2 mRNA contains a cis-repressive element of gene expression. In the present study, we found that the stability of chicken ccn2 mRNA is regulated in a differentiation stage-dependent manner in chondrocytes. We also found that stimulation by bone morphogenetic protein 2, platelet-derived growth factor, and CCN2 stabilized ccn2 mRNA in proliferating chondrocytes but that it destabilized the mRNA in prehypertrophic-hypertrophic chondrocytes. The results of a reporter gene assay revealed that the minimal repressive cis-element of the 3'-UTR of chicken ccn2 mRNA was located within the area between 100 and 150 bases from the polyadenylation tail. Moreover, the stability of ccn2 mRNA was correlated with the interaction between this cis-element and a putative 40-kDa trans-factor in nuclei and cytoplasm. In fact, the binding between them was prominent in proliferating chondrocytes and attenuated in (pre)hypertrophic chondrocytes. Stimulation by the growth factors repressed the binding in proliferating chondrocytes; however, it enhanced it in (pre)hypertrophic chondrocytes. Therefore, gene expression of ccn2 mRNA during endochondral ossification is properly regulated, at least in part, by changing the stability of the mRNA, which arises from the interaction between the RNA cis-element and putative trans-factor.

    DOI: 10.1074/jbc.M411632200

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  • Interaction of AP-1 and the ctgf gene: a possible driver of chondrocyte hypertrophy in growth cartilage. 査読

    Norifumi H Moritani, Satoshi Kubota, Takanori Eguchi, Tomohiro Fukunaga, Takashi Yamashiro, Teruko Takano-Yamamoto, Hideki Tahara, Kazumi Ohyama, Toshio Sugahara, Masaharu Takigawa

    Journal of bone and mineral metabolism21 ( 4 ) 205 - 10   2003年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER-VERLAG TOKYO  

    The expression of the connective tissue growth factor ( ctgf) gene increases along with the differentiation of growth cartilage cells, and the highest expression is observed in the hypertrophic stage. Similarly, recent reports demonstrated c- fos expression in chondrocytes in the early hypertrophic zone of growth cartilage, and suggested that the c- fos gene may play a crucial role in the regulation of hypertrophic differentiation. A chondrocytic human cell line, HCS-2/8, is known to retain a variety of chondrocytic phenotypes. When such cells were kept overconfluent, they expressed increasing levels of c- fos transcripts along a time course phenotypically similar to that of hypertrophic differentiation. Moreover, by using a competitive electromobility-shift assay, we found that AP-1, a Fos/Jun heterodimer, in HCS-2/8 was capable of binding not only to a typical AP-1-binding DNA fragment but also to the enhancer fragment of the ctgf gene. Based on the findings above, we hypothesize that, prior to hypertrophic differentiation, AP-1-related oncogenes are activated and that their gene products subsequently activate ctgf gene expression, which might eventually induce hypertrophy.

    DOI: 10.1007/s00774-003-0410-1

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  • A novel cis-element that enhances connective tissue growth factor gene expression in chondrocytic cells. 査読 国際誌

    Takanori Eguchi, Satoshi Kubota, Seiji Kondo, Takuo Kuboki, Hirofumi Yatani, Masaharu Takigawa

    Biochemical and biophysical research communications295 ( 2 ) 445 - 51   2002年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    To clarify the chondrocyte-specific regulatory mechanism of connective tissue growth factor (ctgf) gene expression, we analyzed the functionality and DNA-protein interaction of the CTGF promoter. Comparative luciferase assay of the CTGF promoter deletion mutants among HCS-2/8 chondrocytic cells and fibroblastic cells revealed that a 110-bp region in the promoter was crucial for the HCS-2/8-specific transcriptional enhancement. Subsequent competitive gel shift assay revealed that transcription factors in HCS-2/8 nuclei bound to a 60-bp portion in the corresponding region. Relative luciferase activity from a CTGF promoter with mutant TGF-beta response element (TbRE) was 16.9% lower than that from an intact promoter. On the other hand, relative luciferase activity from a CTGF promoter with 4bp point mutations at 30bp upstream of the TbRE was 47.7% lower than that from the intact one. The binding activity of HCS-2/8 nuclear factor(s) to the sequence over the 4-bp was remarkably higher than that of any nuclear extract from other types of cells. Therefore, we entitled the sequence 'TRENDIC', a transcription enhancer dominant in chondrocytes, which stands for a novel enhancer for chondrocyte-specific CTGF gene expression.

    DOI: 10.1016/S0006-291X(02)00700-3

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  • Connective tissue growth factor increased by hypoxia may initiate angiogenesis in collaboration with matrix metalloproteinases. 査読 国際誌

    Seiji Kondo, Satoshi Kubota, Tsuyoshi Shimo, Takashi Nishida, Gen Yosimichi, Takanori Eguchi, Toshio Sugahara, Masaharu Takigawa

    Carcinogenesis23 ( 5 ) 769 - 76   2002年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Connective tissue growth factor (CTGF) is known to be a potent angiogenic factor. Here we investigated how CTGF and matrix metalloproteinases (MMPs) are involved in the early stage of hypoxia-induced angiogenesis using human breast cancer cell line, MDA231, and vascular endothelial cells. Hypoxic stimulation (5% O(2)) of MDA231 cells increased their steady-state level of ctgf mRNA by approximately 2-fold within 1.5 h, and the levels remained at a plateau up to 6 h, and then decreased by 12 h as compared with the cells cultured under the normoxic condition. Membrane-type 1 MMP (MT1-MMP) mRNA levels was also increased within a few hours of the exposure to hypoxia. Indeed, ELISA revealed that the CTGF protein/cell in medium conditioned by MDA231 cells exposed to hypoxia was maximally greater at 24 h than in the medium from normoxic cultures and that the secretion rate (supernatant CTGF/cell layer CTGF) increased in a time-dependent manner from 24 to 72 h of hypoxic exposure. Hypoxic induction of CTGF was also confirmed by immunohistochemical analyses. Furthermore, zymogram analysis revealed that the production of active MMP-9 was also induced in MDA231 cells incubated under hypoxic conditions. Finally, we found that recombinant CTGF also increased the expression of a number of metalloproteinases that play a role in the vascular invasive processes and decreased the expression of tissue inhibitors of metalloproteinases by vascular endothelial cells. These findings suggest that hypoxia stimulates MDA231 cells to release CTGF as an angiogenic modulator, which initiates the invasive angiogenesis cascade by modulating the balance of extracellular matrix synthesis and degradation via MMPs secreted by endothelial cells in response to CTGF. This cascade may play critical roles in the hypoxia-induced neovascularization that accompanies tumor invasion in vivo.

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  • Cell-type-specific trans-activation of herpes simplex virus thymidine kinase promoter by the human T-cell leukemia virus type I Tax protein. 査読

    Kubota S, Mukudai Y, Hattori T, Eguchi T, Kondo S, Takigawa M

    DNA and cell biology20 ( 9 ) 563 - 568   2001年9月

  • Novel mode of processing and secretion of connective tissue growth factor/ecogenin(CTGF/Hcs24) in chondrocytic HCS-2/8 cells 査読

    Kubota S, Eguchi T, Shimo T, Nishida T, Hattori T, Kondo S, Nakanishi T, Takigawa M

    Bone29   155 - 161   2001年

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    掲載種別:研究論文(学術雑誌)  

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  • Characterization of a mouse ctgf 3 '-UTR segment that mediates repressive regulation of gene expression

    S Kondo, S Kubota, T Eguchi, T Hattori, T Nakanishi, T Sugahara, M Takigawa

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS278 ( 1 ) 119 - 124   2000年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC  

    We isolated a small segment of the 3'-untranslated region (3'-UTR) in the mouse connective tissue growth factor (ctgf/fisp12) gene and evaluated its functionality. Comparison of the nucleotide sequences of human and mouse ctgf 3'-UTRs revealed a conserved small segment of 91 bases, The corresponding segments of the 3'-UTRs shared as much as 82.4% homology, whereas the overall homology between the 3'-UTRS was 71.8%. To study the functionality of the conserved segment, the corresponding region of mouse ctgf cDNA was amplified from NIH3T3 cells. When it was fused downstream of a marker gene, it showed remarkable repressive effects on gene expression. The repressive effect of the sense form was more prominent than that of the antisense form. Computer analyses of these sequence predicted stable secondary structures, suggesting that they act at the RNA level. The predicted structures of the sense and antisense forms appeared to be slightly different, which is consistent with the difference in repressive function. These findings defined the conserved small element in the mouse ctgf gene as a potent negative regulator of gene expression, which may act at a posttranscriptional level. (C) 2000 Academic Press.

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  • Identification of an RNA element that confers post-transcriptional repression of connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) gene: Similarities to retroviral RNA-protein interactions

    S Kubota, S Kondo, T Eguchi, T Hattori, T Nakanishi, RJ Pomerantz, M Takigawa

    ONCOGENE19 ( 41 ) 4773 - 4786   2000年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    The repressive effect of the 3'-untranslated region (3'-UTR) in human connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) mRNA on gene expression had been demonstrated in our previous study. Here, we identified a minimal RNA element in the 3'-UTR, which acts as a cis-acting element of structure-anchored repression (CAESAR). Deletion analyses of the 3'-UTR led us to minimize the element of 84 bases at the junction of the coding region and the 3'-UTR. The minimized RNA segment is predicted, and actually capable of forming a stable secondary structure in vitro. Mutational analyses disclosed a significant relationship between the predicted structure and repressive effect. The utility of CAESAR as a post-transcriptional regulatory element was represented by the fact that steady-state mRNA levels were not affected by CAESAR linked in cis, while protein levels from such a chimeric gene were markedly reduced. Of note, the CAESAR sequence exerted no effect, when it was placed upstream of the promoter. Finally, RNA gel electromobility-shift analyses demonstrated a nuclear factor that interacts with the folded CAESAR. Taken together, it was uncovered that CAESAR of ctgf is a novel post-transcriptional structured RNA regulatory element, probably acting through direct interactions with a nuclear factor as observed in retroviral RNA elements with certain proteins.

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書籍等出版物

  • HSP Stimulation on Macrophages and Dendritic Cells Activates Innate Immune System. In: . Heat Shock Proteins.

    Yanyin Lu, Takanori Eguchi

    Springer, Dordrecht  2020年 

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  • Heat Shock Proteins and Periodontitis – Cross-Reaction Between Bacterial and Human HSP in Periodontal Infection Linking with Cardiovascular Diseases. In: . Heat Shock Proteins.

    Tadashi Yamamoto, Takanori Eguchi

    Springer, Dordrecht  2020年 

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  • Extracellular Vesicle-Associated Moonlighting Proteins: Heat Shock Proteins and Metalloproteinases. In: . Heat Shock Proteins.

    Takanori Eguchi, Eman A. Taha

    Springer, Dordrecht  2020年 

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  • Roles of HSP on Antigen Presentation. In: . Heat Shock Proteins.

    Kazuyuki Furuta, Takanori Eguchi

    Springer, Dordrecht  2020年 

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  • Regulatory Roles of HSP90-Rich Extracellular Vesicles. In: Asea A., Kaur P. (eds) Heat Shock Protein 90 in Human Diseases and Disorders. Heat Shock Proteins, vol 19.

    Eguchi T, Ono K, Kawata K, Okamoto K, Calderwood S.K

    Springer, Cham  2019年 

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  • Monitoring of the Heat Shock Response with a Real-Time Luciferase Reporter. In: Stuart Calderwood and Thomas Prince. (eds) Chaperones. Methods in Molecular Biology, vol 1709.

    Toshiki Kijima, Takanori Eguchi, Len Neckers, Thomas L. Prince

    Humana Press, New York, NY  2018年 

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  • Regulatory Roles for Hsp70 in Cancer Incidence and Tumor Progression. In: Mario D. Galigniana (ed) Frontiers in Structural Biology, vol 1. Role of Molecular Chaperones in Structural Folding, Biological Functions, and Drug Interactions of Client Proteins.

    Taka Eguchi, Benjamin J. Lang, Ayesha Murshid, Thomas Prince, Jianlin Gong, Stuart K Calderwood

    Bentham Science Publisher  2018年 

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  • Promoter analyses of CCN genes. in Methods in Molecular Biology. CCN Proteins: Methods and Protocols

    Eguchi T, Kubota S, Takigawa M

    Humana Press, New York, NY  2017年1月 

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  • CCN proteins in Health and Diseases: Novel Transcriptional Regulation of CCN2/CTGF by Nuclear Translocation of MMP3

    Eguchi T, Kubota S, Kawata K, Mukudai, Y, Takigawa M( 担当: 共著)

    Springer Dordrecht Heidelberg London New York  2010年1月 

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  • CCN Proteins in Health and Disease: Nucleophosmin/B23: A Multifunctional Regulator that Determines the Fate of CCN2 mRNA.

    Kubota S, Mukudai Y, Kawaki H, Kondo S, Eguchi T, Sumiyoshi K, Ohgawara T, Shimo T, Takigawa M( 担当: 共著)

    Springer Dordrecht Heidelberg London New York  2010年1月 

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  • エンドトキシン研究12:58-60. ヒストンアセチル化制御薬を用いたHMGB1の放出制御

    杉浦進介, 江口傑徳, 小松寿明, 松下健二

    医学図書出版  2009年 

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MISC

  • ニコチンは口腔扁平上皮癌細胞のセツキシマブ耐性を促進する

    清水 理恵子, 伊原木 聰一郎, 江口 傑徳, 奥井 達雄, 高畠 清文, 河合 穂高, 小野 喜章, 岡元 邦彰, 長塚 仁, 佐々木 朗

    岡山歯学会雑誌37 ( 2 ) 80 - 81   2018年12月

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    記述言語:日本語   出版者・発行元:岡山歯学会  

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  • 口腔扁平上皮癌診断・治療における分子シャペロンHSP90含有細胞外小胞の可能性

    小野 喜章, 江口 傑徳, 十川 千春, 奥舎 有加, 河合 穂高, 中野 敬介, 佐々木 朗, 岡元 邦彰, 小崎 健一

    Journal of Oral Biosciences Supplement2018   333 - 333   2018年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • 核内およびエクソソーム中に存在するMMPs/PEXの癌進展における役割とその抑制

    奥舎 有加, 江口 傑徳, 十川 千春, 小野 喜章, 奥井 達雄, 中野 敬介, 岡元 邦彰

    Journal of Oral Biosciences Supplement2018   150 - 150   2018年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • 癌の治療抵抗性と転移におけるHSP90およびMMP3の役割

    江口 傑徳, 小野 喜章, 奥舎 有加, 十川 千春, 内部 健太, 中野 敬介, 奥井 達雄, 滝川 正春, 岡元 邦彰, カルダーウッド・スチュアート

    Journal of Oral Biosciences Supplement2018   142 - 142   2018年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • CATABOLIC EFFECTS OF FGF-1 ON CHONDROCYTES WITH REDUCED CCN2 PRODUCTION THAT PROMOTES CARTILAGE REGENERATION: POSSIBLE ROLE IN OSTEOARTHRITIS

    A. Elseoudi, T. Abd El Kader, T. Nishida, E. Aoyama, T. Eguchi, M. Takigawa, S. Kubota

    OSTEOPOROSIS INTERNATIONAL28   S225 - S225   2017年7月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:SPRINGER LONDON LTD  

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  • An old dog's new tricks: iMMP-3 Processes Nuclear Matrix and Converts Heterochromatin Proteins Leading to Transcriptional Promotion of HSP genes

    Takanori Eguchi, Akihisa Mino, Benjamin J. Lang, Stuart K. Calderwood

    FASEB JOURNAL30   2016年4月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:FEDERATION AMER SOC EXP BIOL  

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  • An old dog's new tricks: iMMP-3 Processes Nuclear Matrix and Converts Heterochromatin Proteins Leading to Transcriptional Promotion of HSP genes

    Takanori Eguchi, Akihisa Mino, Benjamin J. Lang, Stuart K. Calderwood

    FASEB JOURNAL30   2016年4月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:FEDERATION AMER SOC EXP BIOL  

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  • Cellular Reprogramming Using Defined Factors and MicroRNAs

    Takanori Eguchi, Takuo Kuboki

    STEM CELLS INTERNATIONAL   2016年

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:HINDAWI PUBLISHING CORP  

    Development of human bodies, organs, and tissues contains numerous steps of cellular differentiation including an initial zygote, embryonic stem (ES) cells, three germ layers, and multiple expertized lineages of cells. Induced pluripotent stem (iPS) cells have been recently developed using defined reprogramming factors such as Nanog, Klf5, Oct3/4 (Pou5f1), Sox2, and Myc. This outstanding innovation is largely changing life science and medicine. Methods of direct reprogramming of cells into myocytes, neurons, chondrocytes, and osteoblasts have been further developed using modified combination of factors such as N-myc, L-myc, Sox9, and microRNAs in defined cell/tissue culture conditions. Mesenchymal stem cells (MSCs) and dental pulp stem cells (DPSCs) are also emerging multipotent stem cells with particular microRNA expression signatures. It was shown that miRNA-720 had a role in cellular reprogramming through targeting the pluripotency factor Nanog and induction of DNA methyltransferases (DNMTs). This review reports histories, topics, and idea of cellular reprogramming.

    DOI: 10.1155/2016/7530942

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  • Micro RNA 18a regulates chondrocytic phenotype: Involvement of Ccn2/Ctgf as a major target gene

    T. Ohgawara, S. Kubota, H. Kawaki, S. Kondo, T. Eguchi, A. Sasaki, M. Takigawa

    BONE44   S42 - S43   2009年5月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:ELSEVIER SCIENCE INC  

    DOI: 10.1016/j.bone.2009.01.109

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  • Novel transcription factor-like function of MMP-3/stromelysin-1 that regulates connective tissue growth factor (CTGF/CCN2) gene transcription

    T. Eguchi, S. Kubota, K. Kawata, Y. Mukudai, T. Yanagita, T. Ohgawara, J. Uehara, S. Ibaragi, M. Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH22   S261 - S261   2007年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC BONE & MINERAL RES  

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  • Purification and functional characterization of a protein that regulate ccn2 gene expression during chicken chondrocyte differentiation.

    Y. Mukudai, S. Kubota, S. Kondo, T. Eguchi, H. Kawaki, M. Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH21   S149 - S149   2006年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC BONE & MINERAL RES  

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  • Post-transcriptional regulation of CCN2/CTGF gene expression during differentiation of chicken chondrocytes: involvement of a putative trans-factor which interacts with a cis-element in the 3 '-UTR of mRNA

    Y Mukudai, S Kubota, T Eguchi, S Kondo, K Nakao, M Takigawa

    FEBS JOURNAL272   284 - 285   2005年7月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:BLACKWELL PUBLISHING  

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  • Novel cis-element TRENDIC that enhance connective tissue growth factor (ctgf) gene expression in chondrocytic HCS-2/8.

    T Eguchi, S Kubota, S Kondo, Y Mukudai, T Kuboki, H Yatani, M Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH17   S224 - S224   2002年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC BONE & MINERAL RES  

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  • Effects of IL-1beta and LPS on CTGF expression in mouse-derived odontoblast-like cells, MDPC-23.

    W Sonoyama, T Kuboki, T Fujisawa, T Eguchi, J Uehara, H Yatani, M Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH17   S327 - S327   2002年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC BONE & MINERAL RES  

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  • 低酸素によるヒト乳癌細胞における結合組織成長因子CTGF及びマトリクスメタロプロテアーゼの発現誘導

    近藤 誠二, 久保田 聡, 志茂 剛, 西田 崇, 吉道 玄, 江口 傑徳, 菅原 利夫, 滝川 正春

    日本癌学会総会記事60回   183 - 183   2001年9月

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    記述言語:日本語   出版者・発行元:日本癌学会  

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  • Promoter activity determinant of human connective tissue growth factor (CTGF/Hcs24) gene in a human chondrocytic cell line, HCS-2/8.

    T Eguchi, S Kubota, S Kondo, T Shimo, T Nakanishi, T Kuboki, H Yatani, M Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH16   S326 - S326   2001年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC BONE & MINERAL RES  

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  • 低酸素による結合組織成長因子(CTGF)及びマトリクスメタロプロテアーゼ(MMP)活性の協調的発現誘導

    近藤 誠二, 久保田 聡, 志茂 剛, 西田 崇, 吉道 玄, 江口 傑徳, 菅原 利夫, 滝川 正春

    歯科基礎医学会雑誌43 ( 5 ) 632 - 632   2001年8月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • ヒト軟骨様細胞株HCS-2/8におけるCTGF/Hcs24遺伝子のプロモーター活性決定因子

    江口 傑徳, 久保田 聡, 近藤 誠二, 志茂 剛, 中西 徹, 窪木 拓男, 矢谷 博文, 滝川 正春

    歯科基礎医学会雑誌43 ( 5 ) 557 - 557   2001年8月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • 軟骨様細胞株HCS-2/8における多機能成長因子CTGF/Hcs24の転写から分泌まで

    江口 傑徳, 久保田 聡, 志茂 剛, 近藤 誠二, 中西 徹, 矢谷 博文, 滝川 正春

    生化学73 ( 8 ) 778 - 778   2001年8月

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    記述言語:日本語   出版者・発行元:(公社)日本生化学会  

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  • 軟骨由来成長因子CTGF/Hcs24のヒト軟骨細胞株HCS-2/8におけるプロセシングと分泌の様態

    久保田 聡, 江口 傑徳, 志茂 剛, 西田 崇, 服部 高子, 近藤 誠二, 中西 徹, 滝川 正春

    日本骨代謝学会雑誌19 ( 2 ) 104 - 104   2001年7月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • ヒト軟骨肉腫由来軟骨様細胞株HCS-2/8における結合組織成長因子CTGF/Hcs24遺伝子のプロモーター活性決定因子

    江口 傑徳, 久保田 聡, 近藤 誠二, 志茂 剛, 中西 徹, 窪木 拓男, 矢谷 博文, 滝川 正春

    日本骨代謝学会雑誌19 ( 2 ) 102 - 102   2001年7月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • 結合組織成長因子CTGF/Hcs24の軟骨細胞様細胞株HCS-2/8での発現と動態制御

    久保田 聡, 江口 傑徳, 志茂 剛, 服部 高子, 近藤 誠二, 中西 徹, 滝川 正春

    Connective Tissue33 ( 2 ) 157 - 157   2001年6月

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    記述言語:日本語   出版者・発行元:日本結合組織学会  

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  • Regulatory mechanism of human connective tissue growth factor (ctgf/hcs24) gene expression in a human chondrocytic cell line, HCS-2/8.

    T.Eguchi, S.Kubota, S.Kondo, T.Shimo, T.Hattori, T.Nakanishi, T.Kuboki, H.Yatani, M.Takigawa

    Journal of Biochemistry   2001年

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  • A novel RNA element that confers post-transcriptional repression of human connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) gene.

    S Kubota, S Kondo, T Eguchi, T Hattori, T Nakanishi, RJ Pomerantz, M Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH15   S340 - S340   2000年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC BONE & MINERAL RES  

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  • A cis-acting repressive element in the 3 '-untranslated region of the CTGF gene.

    S Kubota, T Hattori, T Eguchi, S Kondo, T Nakanishi, M Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH14   S436 - S436   1999年9月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC BONE & MINERAL RES  

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▼全件表示

講演・口頭発表等

  • 軟骨由来成長因子CTGF/Hcs24のヒト軟骨細胞株HCS-2/8におけるプロセシングと分泌の様態

    久保田 聡, 江口 傑徳, 志茂 剛, 西田 崇, 服部 高子, 近藤 誠二, 中西 徹, 滝川 正春

    日本骨代謝学会雑誌  2001年7月  (一社)日本骨代謝学会

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    開催年月日: 2001年7月

    記述言語:日本語  

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  • 結合組織成長因子CTGF/Hcs24の軟骨細胞様細胞株HCS-2/8での発現と動態制御

    久保田 聡, 江口 傑徳, 志茂 剛, 服部 高子, 近藤 誠二, 中西 徹, 滝川 正春

    Connective Tissue  2001年6月  日本結合組織学会

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    開催年月日: 2001年6月

    記述言語:日本語  

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  • Regulatory mechanism of human connective tissue growth factor (ctgf/hcs24) gene expression in a human chondrocytic cell line, HCS-2/8.

    T.Eguchi, S.Kubota, S.Kondo, T.Shimo, T.Hattori, T.Nakanishi, T.Kuboki, H.Yatani, M.Takigawa

    Journal of Biochemistry  2001年 

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    開催年月日: 2001年

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  • A novel RNA element that confers post-transcriptional repression of human connective tissue growth factor/hypertrophic chondrocyte specific 24 (ctgf/hcs24) gene.

    S Kubota, S Kondo, T Eguchi, T Hattori, T Nakanishi, RJ Pomerantz, M Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH  2000年9月  AMER SOC BONE & MINERAL RES

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    開催年月日: 2000年9月

    記述言語:英語  

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  • A cis-acting repressive element in the 3 '-untranslated region of the CTGF gene.

    S Kubota, T Hattori, T Eguchi, S Kondo, T Nakanishi, M Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH  1999年9月  AMER SOC BONE & MINERAL RES

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    開催年月日: 1999年9月

    記述言語:英語  

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  • オルガノイドと細胞外小胞を癌研究に応用する

    江口傑徳, 十川千春, 岡元邦彰

    第62回歯科基礎医学会学術大会:アップデートシンポジウム「発生と疾患にみる新たな細胞間コミュニケーション」  2020年9月 

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  • 三次元腫瘍オルガノイドの開発と細胞外小胞の新機能

    江口傑徳

    第125回日本解剖学会総会全国学術集会:シンポジウム「顎口腔領域の発生と疾患に見る細胞間情報伝達機構の新たなカタチ」  2020年3月 

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  • 細胞間情報伝達・細胞外小胞研究国際シンポジウム開催報告と今後

    江口傑徳

    第50回 ARCOCSセミナー:次世代研究育成グループ研究成果中間報告  2020年2月 

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  • パーキンソン病治療薬の抗癌作用(ドラッグリポジショニングの提案)

    十川千春, 江口傑徳, Tran Tien Manh, 石毛真行, 河合穂高, 奥舎有加, 中野敬介, 十川紀夫, 小崎健一, 岡元邦彰

    第45回 岡山脳研究セミナー  2020年1月28日 

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  • The Roles of Macrophage-derived Exosomes on the Oral Cancer Cells

    Toshiki Nara, Taka Eguchi, Chiharu Sogawa, Kuniaki Okamoto

    第4回岡山医療教育国際シンポジウム  2019年12月 

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  • Roles of MMP3 in Cargo Signature and Transmissive Activity of Cancer Extracellular Vesicles (Poster Competition Award)

    Eman A Taha, Taka Eguchi, Chiharu Sogawa, Kuniaki Okamoto

    第4回岡山医療教育国際シンポジウム  2019年12月 

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  • Exosome-mediated Intercellular Communication using Multiplexing Fluorescent and Bioluminescent Reporter Systems

    Yanyin Lu, Taka Eguchi, Chiharu Sogawa, Kuniaki Okamoto

    第4回岡山医療教育国際シンポジウム  2019年12月 

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  • オルガノイドを応用したドラッグリポジショニング開発

    十川千春, 江口傑徳, Tran Tien Manh, 石毛真行, 河合穂高, 奥舎有加, 中野敬介, 十川紀夫, 小崎健一, 岡元邦彰

    岡山歯学会  2019年12月 

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  • Tumoroidを応用したゲノム研究の可能性

    江口傑徳

    第2回岡山大学ゲノミクス研究会  2019年11月18日 

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  • 分子シャペロントリオによるエクソソーム制御,腫瘍悪性化およびマクロファージ分極について

    江口 傑徳, 小野 喜章, 河合 穂高, チャン・チェン・マン, 十川 千春, 奥舎 有加, 岡元 邦彰

    日本臨床ストレス応答学会  2019年11月3日 

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  • Oncosome and Tumoroid

    Takanori Eguchi

    細胞間情報伝達・細胞外小胞研究会第一回国際シンポジウム  2019年11月 

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  • Cancer exosomes initiate EMT involving drug resistance and tumorigenesis.

    Takanori Eguchi, Kisho Ono, Chiharu Sogawa, Yuka Okusha, Eman Ahmed Taha, Manh Tien Tran, Yanyin Lu, Kuniaki Okamoto

    The 9th International Epithelial-Mesenchymal Transition International Association meeting (TEMTIA IX), the 35th International Kumamoto Medical Bioscience Symposium  2019年11月 

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  • Roles of MMP3-containing extracellular vesicles in tumorigenesis, CD9-exosome release, and cell-cell communication in cancer

    Eman A Taha, Yuka Okusha, Chiharu Sogawa, Abdellatif El-Seoudi, Satoshi Kubota, Ayano Satoh, Kuniaki Okamoto, Taka Eguchi

    細胞間情報伝達・細胞外小胞研究会第一回国際シンポジウム  2019年11月 

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  • Rab11A negatively regulates osteoclastogenesis through modulating the transport route of cell surface receptors

    Yuka Okusha, Mami Itagaki, Manh Tien Tran, Chiharu Sogawa, Takanori Eguchi, Tatsuo Okui, Kuniaki Okamoto

    細胞間情報伝達・細胞外小胞研究会第一回国際シンポジウム  2019年11月 

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  • Monitoring of exosome using multiplexing fluorescent and bioluminescent reporter systems

    Yanyin Lu, Toshiki Nara, Chiharu Sogawa, Eman A Taha, Noa Matsuda, Kuniaki Okamoto, Takanori Eguchi

    細胞間情報伝達・細胞外小胞研究会第一回国際シンポジウム  2019年11月 

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  • Drug repositioning using the three-dimensional tumor organoid monitoring system for anti-cancer drug screening

    Chiharu Sogawa, Takanori Eguchi, Masayuki Ishige, Hotaka Kawai, Keisuke Nakano, Yuka Okusha, Norio Sogawa, Ken-ichi Kozaki, Kuniaki Okamoto

    細胞間情報伝達・細胞外小胞研究会第一回国際シンポジウム  2019年11月 

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  • 口腔扁平上皮癌診断・治療における分子シャペロンHSP90含有エクソソームの可能性

    小野喜章, 江口傑徳, 中野敬介, 河合穂高, 佐々木朗

    第64回公益社団法人日本口腔外科学会総会・学術大会  2019年10月 

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  • 新しい腫瘍オルガノイド多元評価システムの開発

    十川千春, 江口傑徳, 大山和美, 奥舎有加, 中野敬介, 十川紀夫, 岡元邦彰

    第61回日本歯科基礎医学会  2019年10月 

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  • 口腔癌の進展と抵抗性における細胞外小胞の重要な役割

    江口傑徳

    第61回日本歯科基礎医学会:アップデートシンポジウム「がんの多様性と新規治療法への展望」  2019年10月 

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  • 口腔癌エクソソームによるマクロファージ分極および腫瘍悪性化における分子シャペロントリオの重要性

    江口 傑徳, 小野 喜章, 河合 穂高, チャン・チェン・マン, 十川 千春, 奥舎 有加, 岡元 邦彰

    第6回 日本細胞外小胞学会  2019年10月 

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  • 悪性腫瘍の微小環境におけるCDC37とHSP90の重要性

    江口傑徳

    第36回日本ハイパーサーミア学会:シンポジウム「Heat Shock Proteinとがん温熱療法の最前線」  2019年9月6日 

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  • 高転移性癌細胞で高発現するMMP3が有するCtgf/Ccn2発現調節機能と細胞外小胞の関連

    奥舎 有加, 江口 傑徳, タハ・エマン, チャン・チェン・マン, 十川 千春, 青山絵里子, 滝川正春, 岡元 邦彰

    第11回日本CCNファミリー研究会  2019年8月31日 

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  • Roles of MMP3-containing extracellular vesicles in tumorigenesis, CD9-exosome release, and cell-cell communication in cancer

    第11回日本CCNファミリー研究会  2019年8月31日 

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  • 口腔癌エクソソームによる腫瘍悪性化及びマクロファージM2分極における分子シャペロントリオCDC37/HSP90の重要性と標的性

    江口傑徳, 小野喜章, 河合穂高, チャン・チェン・マン, 十川千春, 奥舎有加, 岡元邦彰

    第11回 日本RNAi研究会  2019年8月28日 

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  • 細胞外小胞の積荷・伝達機能および腫瘍形成におけるMMP3の役割

    エマン・タハ, 江口傑徳, 奥舎有加, 十川千春, アブデラティフ・エルソウディ, 久保田聡, 佐藤あやの, 岡元邦彰

    第11回 日本RNAi研究会  2019年8月28日 

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  • Anti-tumor effect by regulation of HSP90 / CDC37

    Takanori Eguchi

    Seminar at Sunshine Hospital Victoria University, Melbourne  2019年3月29日 

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  • How do Exosomes promote Carcinogenesis?

    Takanori Eguchi

    Special Cancer Biology Seminar at QIMRB Medical Research Institute, Brisbane  2019年3月27日 

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    記述言語:英語  

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  • Exosomal and Transcriptional Roles of MMP in Cancer

    Takanori Eguchi

    Special Guest Lecture at the University of Sydney  2019年3月25日 

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    記述言語:英語  

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  • 脂質・コレステロール排出ポンプABCG1標的による腫瘍内エクソソーム蓄積および腫瘍縮小

    江口 傑徳, 十川 千春, 難波 友里, 奥舎 有加, 河合 穂高, 小野 喜章, 板垣 まみ, 村上 純, 大山 和美, 浅海 淳一, 岡元 邦彰

    第92回日本薬理学会  2019年3月 

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    記述言語:英語   会議種別:口頭発表(一般)  

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  • マトリックス・メタロプロテアーゼ(MMPs)の古典的機能と新機能

    江口 傑徳

    BioScience Retreat in Tosa 2017  2018年12月3日 

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  • Tumoroid Never Knows? 腫瘍オルガノイドと細胞外小胞からみる難治性がん研究・治療のミライ

    江口 傑徳

    第41回日本分子生物学会年会  2018年11月30日 

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    記述言語:英語  

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  • RASP: 癌のストレス抵抗性と小胞/HSP分泌特性

    江口 傑徳

    2018 Bioscience Retreat in Awaji  2018年11月24日 

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    記述言語:英語  

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  • Secretion of Extracellular Vesicles with Heat Shock Proteins by Resistant Adenocarcinomas and Metastatic Oral Cancer

    江口 傑徳

    Cold Spring Harbor Asia  2018年11月16日 

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    記述言語:英語  

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  • がん幹細胞特性をもつオルガノイドによるEPCAMエクソソームおよびHSP90の分泌

    江口 傑徳

    日本癌学会学術総会 第77回  2018年9月28日 

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  • 大腸癌の増殖,浸潤,転移を促進するPEXドメインの発見とその抑制

    江口 傑徳

    The intranuclear PEX domain of MMP involves proliferation, migration, and metastasis of aggressive adenocarcinoma cells  2018年9月28日 

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    記述言語:英語  

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  • 抗体医薬耐性および上皮間葉転換におけるがん細胞外小胞の役割: Roles for cancer EVs in antibody therapeutic resistance and epithelial-to-mesenchymal transition

    Eguchi T, Ono K, Fujiwara T, Sogawa C, Calderwood SK, 江口傑徳, 小野喜章, 藤原敏史, 十川千春, スチュアート, カルダーウッド

    第10回 日本RNAi研究会 第5回 日本細胞外小胞学会  2018年8月30日 

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  • 癌と軟骨,その両者において鍵を握るMMP-CCN制御軸: MMP-CCN Regulatory Axis is a Common Key to Cancer and Cartilage Metabolisms. (シンポジウム)

    江口 傑徳

    日本CCNファミリー研究会 第10回記念大会  2018年8月25日 

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  • 癌の治療抵抗性と転移におけるHSP90およびMMP3の役割

    江口傑徳, 小野喜章, 奥舎有加, 十川千春, 内部健太, 中野敬介, 河合穂高, 奥井達雄, 岡元邦彰, スチュアート, カルダーウッド

    歯科基礎医学会 第60回  2018年 

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  • RASP: 難治性がんにみられるストレス抵抗性と小胞/HSP分泌特性(シンポジウム)

    江口 傑徳

    臨床ストレス応答学会  2018年 

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    会議種別:シンポジウム・ワークショップ パネル(指名)  

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  • エクソソームの基礎,最新知見,展望

    江口 傑徳

    ARCOCSセミナー  2018年 

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  • がんをなくす:New Approaches Revealing Novel Secretory Phenotype, Pathology, and Therapeutic Targets in Resistant / Refractory Cancer (ベストプレゼンテーション賞受賞)

    江口 傑徳

    ブレインストーミング2018  2018年 

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  • 癌や炎症性疾患において分子シャペロンHSPsが誘導される新たな経路: 核内MMPとヘテロクロマチンタンパク質の相互作用

    江口 傑徳

    第12回 臨床ストレス応答学会  2017年11月5日 

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  • 細胞内におけるマトリックスメタロプロテアーゼ(MMP)の役割

    江口 傑徳

    歯科基礎医学会  2017年9月17日 

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  • MMP3はヘテロクロマチンタンパク質HP1と相互作用してHSP遺伝子群を制御する

    江口 傑徳

    2017年8月26日 

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  • 三次元培養技術を用いた創薬の試み

    江口 傑徳

    ARCOCS seminar  2017年2月15日 

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  • 「トランスオミックスによる癌細胞耐性メカニズム解明への挑戦 」

    江口 傑徳

    第4回次世代がんインフォマティクス研究会  2016年12月16日 

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  • 転写因子MZF1とSCAND1による前立腺癌細胞の上皮間葉転換、分子シャペロンCDC37発現、キナーゼシグナルの制御

    江口 傑徳

    第39回 日本分子生物学会  2016年11月30日 

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  • 癌細胞ストレス耐性における細胞内MMP3の役割

    江口 傑徳

    第37回岡山歯学会学術集会  2016年10月16日 

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  • 転写因子MZF1とSCAND1は前立腺癌細胞においてEMT,癌促進性キナーゼシグナルおよび分子シャペロン発現を制御する.

    江口 傑徳

    第75回日本癌学会学術総会  2016年10月6日 

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  • OstemiR: A Novel Panel of MicroRNA Biomarkers in Osteoblastic and Osteocytic Differentiation from Mesencymal Stem Cells.

    江口 傑徳

    米国生化学分子生物学会・実験生物学学会  2015年3月 

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  • Intra-nuclear MMP-3 controls transcription of HSP70 gene through interaction with heterochromatin proteins

    江口 傑徳

    米国生化学分子生物学会・実験生物学学会  2015年3月 

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  • 転写コンポーネントによる分子シャペロンおよび腫瘍の制御(講演) 招待

    江口 傑徳

    Molecular Biology of Calcified Tissueセミナー / BioForum@Dental School 合同講演会  2014年3月18日 

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  • 間葉系幹細胞の骨細胞分化における新規の マイクロRNAバイオマーカー(講演)

    江口 傑徳

    Molecular Biology of Calcified Tissueセミナー  2013年6月3日 

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  • Control of Cdc37/Hsp90 and kinase signaling in prostate cancer by SCAN domain proteins. 招待

    江口 傑徳

    国際ハイパーサーミア腫瘍学会第11回大会・日本ハイパーサーミア学会第29回大会合同大会  2012年8月 

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    記述言語:英語  

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  • 岡山歯学会賞受賞講演

    江口 傑徳

    2009年11月8日 

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  • 日本軟骨代謝学会賞受賞講演

    江口 傑徳

    2009年3月7日 

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  • Novel transcriptional Regulation of CCN2/CTGF by Nuclear Translocation of MMP3 (シュプリンガー スカラシップ 受賞講演)

    江口 傑徳

    第5回国際CCN研究会  2008年10月22日 

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▼全件表示

受賞

  • 日本軟骨代謝学会賞

    2009年  

    江口傑徳

     詳細を見る

  • International CCN Society Springer Scholarship

    2008年  

    Takanori Eguchi

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  • 岡山歯学会優秀論文賞

    2020年11月  

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  • プレインストーミング2018最優秀発表賞

    2018年9月  

    江口 傑徳

     詳細を見る

  • 生物学研究奨励賞

    2016年10月   両備檉園記念財団  

    江口 傑徳

     詳細を見る

  • 医学系研究奨励

    2009年11月   武田科学振興財団  

    江口 傑徳

     詳細を見る

  • 岡山歯学会奨励論文賞

    2009年  

    江口傑徳

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▼全件表示

共同研究・競争的資金等の研究

  • コンドロニュートリゲノミクス研究の開拓とニュートリジェネティクス研究への展開

    研究課題/領域番号:20K20611  2020年07月 - 2024年03月

    日本学術振興会  科学研究費助成事業 挑戦的研究(開拓)  挑戦的研究(開拓)

    滝川 正春, 青山 絵理子, 星島 光博, 久保田 聡, 西田 崇, 江口 傑徳

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    配分額:25870000円 ( 直接経費:19900000円 、 間接経費:5970000円 )

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  • 口腔扁平上皮癌における新規遠隔転移抑制治療の開発

    研究課題/領域番号:20H03888  2020年04月 - 2025年03月

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    長塚 仁, 山近 英樹, 中野 敬介, 河合 穂高, 江口 傑徳, 辻極 秀次, 高畠 清文

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    配分額:17030000円 ( 直接経費:13100000円 、 間接経費:3930000円 )

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  • 三次元腫瘍オルガノイド評価系により見出された新規癌転移抑制化合物の創薬展開

    研究課題/領域番号:20K09904  2020年04月 - 2023年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    十川 千春, 岡元 邦彰, 江口 傑徳, 河合 穂高, 青山 絵理子

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    配分額:4420000円 ( 直接経費:3400000円 、 間接経費:1020000円 )

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  • CCNsの細胞内新機能~細胞外新情報伝達系の解明と共通分子基盤の確立及びその応用

    研究課題/領域番号:19H03817  2019年04月 - 2023年03月

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    滝川 正春, 青山 絵理子, 星島 光博, 久保田 聡, 西田 崇, 江口 傑徳, 大野 充昭, 鈴木 守

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    配分額:17290000円 ( 直接経費:13300000円 、 間接経費:3990000円 )

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  • 歯周病原菌が放出する小胞の組織障害性と病態評価への応用

    研究課題/領域番号:19H04051  2019年04月 - 2023年03月

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    岡村 裕彦, 江口 傑徳, 宝田 剛志, 吉田 賀弥, 池亀 美華, 江國 大輔, 伊原木 聰一郎

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    配分額:17160000円 ( 直接経費:13200000円 、 間接経費:3960000円 )

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  • 癌間質による癌の生物学的性格の制御および癌誘発の検討

    研究課題/領域番号:18K09789  2018年04月 - 2021年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    中野 敬介, 長塚 仁, 高畠 清文, 河合 穂高, 江口 傑徳, 辻極 秀次, 志茂 剛

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    資金種別:競争的資金

    配分額:4550000円 ( 直接経費:3500000円 、 間接経費:1050000円 )

    本研究の目的は、口腔癌の間質が癌細胞に作用して、癌の生物学的性質を変化させることの検証、および、癌間質が癌の周囲の正常細胞に働きかけて癌化を引き起こす可能性を検証することである。平成30年度では口腔扁平上皮癌細胞株と間質細胞の組合せによる培養、腫瘍間質の違いによる腫瘍細胞の生物学的性格(増殖能、浸潤能、遊走能、形態)に与える影響を検討した。腫瘍細胞株は高浸潤癌:HSC-2、低浸潤癌:SAS、対照細胞: HaCaT細胞を用いた。間質組織は岡山大学病院手術摘出材料から高浸潤癌症例、低浸潤癌症例を選定して用いた。由来の異なる癌間質を癌細胞株と共培養し、また同複合体をマウス皮下に移植した。その結果、腫瘍組織は癌の間質の違いによって大きくその性状を変化させることが示された。培養系を用いた検討では共培養する間質細胞の違いにより癌細胞に対する遊走、増殖能が変化することが明らかになった。癌細胞と間質複合体の移植実験では組織学的および免疫組織化学的検討を行い、癌細胞の増殖と分化の様相が間質細胞によって影響されることが明らかになった。また、移植実験系では当初予想していなかった非常に特殊な間質細胞の動態が明らかになった。この点については次年度以降さらに追及していく予定である。本年度の研究結果は、癌間質が直接的に癌の生物学的性格を調節していることを組織学的に明らかにしている。この研究結果は癌間質に注目した癌の治療戦略に結び付く非常に重要な成果である。

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  • 難治性がん克服のための腫瘍オルガノイドを利用した創薬研究

    2018年 - 2019年

    公益財団法人鈴木謙三記念医科学応用研究財団  生活習慣病における医学,薬学の萌芽的研究 

    江口 傑徳

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    担当区分:研究代表者  資金種別:競争的資金

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  • 腫瘍内不均一性を反映する新たな薬剤感受性評価システムの構築

    研究課題/領域番号:17K11669  2017年04月 - 2021年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    大山 和美, 十川 千春, 江口 傑徳, 奥舎 有加

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    資金種別:競争的資金

    配分額:4550000円 ( 直接経費:3500000円 、 間接経費:1050000円 )

    近年,上皮間葉転換(EMT)や腫瘍内に存在するEpCAM陽性の癌幹細胞様細胞(CIC)およびエクソソームが,薬剤耐性等の腫瘍抵抗性に関与することが示唆されている.本課題では,口腔扁平上皮癌(OSCC),去勢抵抗性前立腺がん(CRPC)および転移性大腸がん(MCRC)における腫瘍抵抗性に着目し,「腫瘍内不均一性を反映する新たな薬剤感受性評価システム構築」を進めている.H30年度は,① 転移性の異なるOSCC細胞株に由来するエクソソームのプロテオミクスを実施し,がん細胞生存因子HSP90AlphaおよびHSP90Betaを同定し,臨床検体データベース解析により予後予測バイオマーカーとしての可能性を示した. ② OSCC細胞に由来するエクソソームは,OSCC細胞のEMTを促進しただけでなく,正常口腔上皮細胞株のEMTをも誘導したため,エクソソーム中に癌化因子が含まれると考えられた.OSCCエクソソーム誘導性EMTは,抗EGFR抗体薬セツキシマブによって一部阻害できたものの,セツキシマブがOSCCエクソソームとともに分泌されるという新規薬剤耐性機構を発見した.③ OSCCとCRPCに共通の腫瘍抵抗性分子基盤として,細胞外HSP90の分泌およびEpCAMエクソソームの増加が判明した.④ 単層培養と比較し,生体内腫瘍微小環境を如実に再現しうる腫瘍オルガノイドの作製を進めている.MCRCオルガノイドでは多剤耐性遺伝子ABCG2 / BCRP発現上昇と同時に脂質排出ポンプABCG1の発現上昇を認め,ABCG1によるエクソソーム脂質排出はMCRCの抵抗性および腫瘍形成能を上昇させることが判明した.またマイクロアレイデータベース解析から,ABCG1発現上昇がCRPC,OSCC,および乳癌の予後不良と相関し,予測因子として有用である可能性を示唆した.

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  • 癌遺伝子プロモーター活性を指標とした新規スクリーニング系による既存薬再開発

    研究課題/領域番号:17K11643  2017年04月 - 2021年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    十川 千春, 江口 傑徳, 奥舎 有加, 大山 和美, 中野 敬介, 岡元 邦彰, 十川 紀夫

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    資金種別:競争的資金

    配分額:4550000円 ( 直接経費:3500000円 、 間接経費:1050000円 )

    癌の転移は、原発巣からの浸潤、血管内への侵入、転移先臓器での定着等の過程を経て成立する。本研究では、癌の浸潤・転移過程における腫瘍微小環境変化、すなわち上皮間葉転換、基底膜細胞外マトリックス(ECM)分解、腫瘍血管形成を統合的に評価できる新規薬剤スクリーニング系を開発することを目的とする。これまでに、癌促進性プロテアーゼであるMMP9発現機構に着目し、MMP9高発現肺転移性マウス大腸癌由来LuM1細胞にMMP9プロモーター蛍光レポーター細胞(LuM1/m9)を構築した。平成30年度は、三次元培養および培地成分等の影響で腫瘍オルガノイド形成が異なってくるとの知見(Eguchi T et al, 2018, Plos One)を手掛かりに、LuM1/m9細胞三次元培養下でMMP9発現および腫瘍形成能を多元評価できるシステムを構築した。Wnt/β-catenin経路は腫瘍形成性幹細胞シグナルとして知られているが、同時に、マウスMMP9プロモーター領域にβ-catenin/TCF/LEF結合部位が複数存在することが明らかとなった。さらに、Wnt/β-catenin経路を刺激するLiClをLuM1/m9に作用させたところ、MMP9発現のみならず腫瘍オルガノイド形成を促進することがわかった。また、本システムを利用して、大腸癌治療の第一選択薬である5-フルオロウラシル(5-Fu)を評価した結果、5-Fuは腫瘍オルガノイド形成を阻害したが、MMP9発現を阻害しなかった。さらに、これまでに腫瘍オルガノイド形成阻害を確認している薬剤群について、本システムを用いて評価した結果、腫瘍オルガノイド形成とMMP9発現の両方を阻害する複数の薬剤が見出された。

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  • MZF1誘導性の新規がん幹細胞ストレス耐性に関する検証

    研究課題/領域番号:17K11642  2017年04月 - 2021年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    江口 傑徳, 十川 千春, 奥舎 有加, 岡元 邦彰

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    担当区分:研究代表者  資金種別:競争的資金

    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    新規がん幹細胞ストレス耐性:
    癌の転移や再発の原因と目されるがん幹細胞は,三次元環境下でスフェロイド(球状細胞塊)を形成する.今回試行した67種類の癌細胞株の殆どがスフェロイドを形成したのに対し,8種類は葡萄様細胞塊(GLA)を形成し,そのうち7種類は腺癌由来であった.このGLA形態は癌細胞の新規形態学的特性と考えられた.GLA形成性細胞株PC-3は去勢抵抗性前立腺癌と神経内分泌前立腺癌の特徴を併せ持つ.PC-3細胞は他の癌種と比べてin vivo腫瘍形成能およびリンパ節転移性に優っていた.幹細胞専用培地および三次元培養環境によって,PC-3細胞のGLA形成促進,細胞塊融合によるオルガノイド形成促進,増殖抑制,幹細胞マーカー(CD44v8-10)発現促進,上皮性マーカー(Eカドヘリン,EpCAM,ESRP1/2)発現促進,がん遺伝子発現促進を認めた.他方,二次元培養環境および血清刺激はこれらを抑制した.上記の幹細胞性誘導環境は,がん幹細胞マーカーEpCAM発現を誘導し,EMT誘導転写因子ZEB1の発現を抑制した.二次元培養下PC-3と比べ,PC-3オルガノイドは,タンパク質貯蔵が豊富で,盛んにEpCAM含有エクソソームを分泌し,顕著にストレス応答タンパク質HSP90αを分泌した.治療抵抗性がんは,以上のがん幹細胞特性より,ストレス耐性および治療抵抗性を発揮すると考えられた.
    Takanori Eguchi, Chiharu Sogawa, Yuka Okusha, Kenta Uchibe et al "Organoids with cancer stem cell-like properties secrete exosomes and HSP90 in a 3D nanoenvironment." PLoS One. 2018 Feb 7;13(2):e0191109.

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  • フラボノイドをベースにした抗がん作用をもつサプリメントの開発

    2017年

    基盤C 

    岡元 邦彰、江口傑徳、十川千春

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    資金種別:競争的資金

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  • エクソソームを指標とする体液診断型口腔癌予後予測法の開発

    研究課題/領域番号:16K11722  2016年04月 - 2020年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    村上 純, 岡田 俊輔, 竹中 文章, 江口 傑徳

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    資金種別:競争的資金

    配分額:4550000円 ( 直接経費:3500000円 、 間接経費:1050000円 )

    抗EGFRモノクローナル抗体であるセツキシマブは、癌細胞のEGFRと結合することで抗癌作用を示す有用な抗癌薬であるが、セツキシマブ耐性症例が報告されている。近年、癌細胞が放出する細胞外小胞(EV)が抗癌剤耐性に関与することが明らかとなってきたが、口腔癌の放出するEGFR含有細胞外小胞(EGFR-EV)とセツキシマブの関連性についての報告はほとんどない。そこで、口腔癌細胞株HSC-3が放出するEGFR-EVがセツキシマブ排出に関与している可能性を考え、HSC-3が放出するEVとセツキシマブの関連性について研究を行った。
    HSC-3を無血清培地で培養し、EGFおよびセツキシマブを添加した条件で行った。上記の培養上清からEV抽出試薬を用いてEVを回収し、回収したEVを粒子径分布解析、電子顕微鏡解析およびウェスタンブロッティング(WB)解析により評価した。セツキシマブの抗癌作用に関しては細胞形態変化および悪性性質変化を検討した。粒子径解析では、160nmの範囲に粒子が存在し、電子顕微鏡解析では脂質二重膜に囲まれた球形の粒子が確認でき、EVの定義と一致した。HSC-3ではEGF刺激によりEGFR-EVの放出が促進された。セツキシマブを投与すると、EV放出量に変化は見られず、EV画分にセツキシマブが検出された。細胞形態変化および悪性性質変化が誘導されたHSC-3にセツキシマブを投与すると、細胞形態変化および性質変化は認めず、効果は不完全であった。同条件の上清からEVを回収し、WB解析を行ったところ、EV画分にセツキシマブが検出された。
    上記の結果から、HSC-3はEVを介してセツキシマブを排出する可能性が示唆された。本研究は、口腔癌細胞のセツキシマブ排出機序の一部を解明した初めての報告であり、今後の癌治療における臨床応用への手がかりとなる重要な知見を含んだ内容である。

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  • フラボノイドをベースにした抗がん作用をもつサプリメントの開発

    研究課題/領域番号:16K11863  2016年04月 - 2019年03月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    岡元 邦彰, 坂井 詠子, 西下 一久, 筑波 隆幸, 十川 千春, 江口 傑徳, 奥舎 有加

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    配分額:4680000円 ( 直接経費:3600000円 、 間接経費:1080000円 )

    in vitroではいくつかのフラボノイドには抗がん作用があることが報告されているが、がんの骨浸潤に重要な役割をする破骨細胞や骨芽細胞に関しては検討されていない。今回行った研究において、抗がん作用を持つだけでなく、破骨細胞形成にも関与するフラボノイドをその構造から6種類に分類し解析した。その結果、漢方薬の甘草に含まれるリクイリチゲニンが破骨細胞抑制効果を持ち、骨芽細胞分化への抑制はなく、さらに副作用も少ない構造を持つ薬物であることが明らかとされた。

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  • MMPを介した腫瘍細胞リプログラミングの制御の試み

    2016年 - 2017年

    公益財団法人両備檉園記念財団 

    江口 傑徳

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    担当区分:研究代表者  資金種別:競争的資金

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  • 癌における細胞内MMP3の役割

    2015年01月 - 2015年12月

    ハーバード大学医学部 JCRT  JCRT grant 

    江口 傑徳

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    担当区分:研究代表者  資金種別:競争的資金

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  • Research and Development of Periodontal Bone-Regenerative Method and Agent

    2010年

    MHLW  Incentive for young scientists 

    EGUCHI Takanori

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    資金種別:競争的資金

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  • 歯周病治療薬と歯槽骨再生方法の開発

    2010年

    厚労省  創薬基盤推進研究事業 

    江口 傑徳

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    担当区分:研究代表者  資金種別:競争的資金

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  • 機能性バイオナノ粒子を応用したエキソサイトーシス制御による歯周病治療の戦略

    研究課題/領域番号:21659436  2009年 - 2011年

    日本学術振興会  科学研究費助成事業 挑戦的萌芽研究  挑戦的萌芽研究

    松下 健二, 江口 傑徳

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    配分額:3360000円 ( 直接経費:3000000円 、 間接経費:360000円 )

    歯肉上皮細胞における細胞内輸送関連因子を探索するとともに、その制御の可能性について検討した。その結果、同細胞における物質輸送に重要なタンパク質としてRab5を同定するとともに、同分子に結合する分子としてVinculinを同定した。歯肉上皮細胞へのPorphyromonas gingivalisの侵入にRab5とICAM-1が関わっていることが明らかになった。さらに、P. gingivalisによる血管内皮細胞からのエキソサイト.シス制御にE-selectinが重要であることが明らかになった。

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  • 安定型マトリックスメタロプロテナーゼ3を用いた新しい歯髄炎治療薬の開発

    研究課題/領域番号:21390512  2009年 - 2011年

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    中村 洋, 中島 美砂子, 中田 和彦, 江口 傑徳

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    配分額:18070000円 ( 直接経費:13900000円 、 間接経費:4170000円 )

    感染の程度が異なるイヌ不可逆性歯髄炎モデルを確立し、MMP-3の効果を検討した。MMP-3処理後14日目で血管と神経を伴う歯髄組織の再生が観察され、28日目で象牙質マトリックス形成がみられた。未処理の歯髄組織は壊死していた。MMP-3処理群では3~ 7日目でコントロール群と比較してマクロファージや抗原提示細胞の浸潤は減少し、またIL-6の発現が有意に低下していた。これらの結果はMMP-3の抗炎症作用を示唆しており、MMP-3が新しい不可逆性歯髄炎の治療薬の候補として有望であると考えられた。

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  • 長寿社会への布石: 水平性骨吸収に対する歯槽骨再生方法の開発

    2009年

    武田科学振興財団  医学系研究奨励(生活習慣病) 

    江口傑徳

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    担当区分:研究代表者  資金種別:競争的資金

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  • Fuseki for Longevity Society: Development of Bone Regenerative Medicine Targetting Holizontal Loss of the Periodontal Bone

    2009年

    Takeda Foundation  Incentive for young scientists 

    EGUCHI Takanori

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    資金種別:競争的資金

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  • 安定型マトリックスメタロプロテナーゼ3を用いた新しい歯髄炎治療薬の開発

    2009年

    文科省  科研費 基盤研究B 

    中村洋

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    資金種別:競争的資金

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  • 歯髄幹細胞を用いた象牙質・歯髄再生医療によるウ蝕・歯髄疾患等のための治療技術の開発

    2009年

    国立長寿医療センター  長寿委託費 

    中島美砂子

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    資金種別:競争的資金

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  • Evaluation of Physiological and Pathological Properties of MMP via Molecular Interactions

    2008年 - 2010年

    MEXT  Grant in aid for young scientists (B) 

    EGUCHI Takanori

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    資金種別:競争的資金

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  • 新規分子間相互作用を介したMMPの生理的・病理的機能発現機序の解明

    2008年 - 2009年

    文科省  科研費 若手研究 (B) 

    江口傑徳

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    担当区分:研究代表者  資金種別:競争的資金

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  • 新規分子間相互作用を介したMMPの生理的・病理的機能発現機序の解明

    研究課題/領域番号:20791378  2008年 - 2009年

    日本学術振興会  科学研究費助成事業 若手研究(B)  若手研究(B)

    江口 傑徳

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    配分額:4160000円 ( 直接経費:3200000円 、 間接経費:960000円 )

    マトリックス・メタロ・プロテアーゼ3(MMP3)は細胞外マトリックスや成長因子、そのレセプターを切断することで、組織リモデリング、血管新生、創傷治癒、骨・軟骨リモデリングに関与し、病理的にはリウマチのマーカーでもある。そこにきて我々は、MMP3が核移行し、転写因子としてCTGF/CCN2遺伝子の発現を誘導することを見出した。今回、細胞内で発現させたMMP3およびその機能ドメインであるPEXドメインおよびCatalyticドメインを細胞内で発現させその機能を解析した。全長MMP3およびそれらの変異体は細胞死を誘導することなく一部は核移行した。さらにはマイクロアレイによって、細胞内MMP3の特異的な新しいターゲット遺伝子としてHSP70B'を見出した。細胞内MMP3によってHSP70B'が誘導され、細胞がUV、感染、メカニカル・ロードなどのストレスに対して耐性を獲得すると考えられる。

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  • Identification and characterization of cartilage-specific transcription factor

    2003年 - 2006年

    JSPS  Incentive for JSPS postdoc 

    EGUCHI Takanori

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    資金種別:競争的資金

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  • 軟骨細胞特異的な新規転写因子の単離とその機能解析

    2003年 - 2006年

    日本学術振興会  特別研究員奨励費 

    江口傑徳

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    担当区分:研究代表者  資金種別:競争的資金

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  • 軟骨細胞特異的な新規転写因子の単離とその機能解析

    研究課題/領域番号:03J02535  2003年 - 2005年

    日本学術振興会  科学研究費助成事業 特別研究員奨励費  特別研究員奨励費

    江口 傑徳

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    配分額:3300000円 ( 直接経費:3300000円 )

    内軟骨性骨化、乳癌の骨転移、組織再生などへの強い関与が報告されているタンパク質CCN2/CTGFは、それら生命現象の実験的モデルとして多用されている細胞株HCS-2/8やMDA231でもまた高発現しており、そのメカニズムの一つがCCN2プロモーター上のエンハンサーTRENDICとタンパク質(複合体)の相互作用であることは既に我々が論文報告している。その後TRENDIC結合因子を同定するために行ったサウスウェスタンスクリーニングにより得られたcDNAの1つは細胞外基質分解酵素として知られるMMP-3をコードしていた。そこでMMP-3による転写調節について研究を行っている。
    1.軟骨組織および培養軟骨細胞様細胞株HCS-2./8の免疫染色では、MMP-3は細胞核に局在化していた。
    2.HCS-2/8の細胞成分を核・細胞質に分画し、ウェスタンブロッティングにより検討したところ、両方の画分にMMP-3が検出された。
    3.クロマチン免疫沈降により、通常培養条件下のHCS-2/8細胞においてMMP-3が染色体上のCCN2のプロモーター領域(TRENDICを含む)に結合することが分かった。
    4.TRENDICと核タンパク質複合体の結合は抗MMP-3抗体によって阻害されるというスーパーシフトアッセイの結果からも、MMP-3がTRENDICに結合することが分かる。
    5.各種細胞株で強制発現したMMP-3は細胞質および核に局在した。この強制発現MMP-3によるCCN2プロモーターの活性化は特定の細胞株でのみ認められた。
    6.HCS-2/8におけるCCN2のプロモーター活性はMMP-3インヒビターの添加により抑制された。
    7.質量分析装置を応用することでMMP-3結合性転写調節因子・核輸送因子を同定した。
    現在上記の結果を統合し、論文発表に向け準備中である。

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担当授業科目

  • 歯科薬理学I(演習・実習) (2020年度) 特別  - その他

  • 歯科薬理学I(講義・演習) (2020年度) 特別  - その他

  • 歯科薬理学II(演習・実習) (2020年度) 特別  - その他

  • 歯科薬理学II(講義・演習) (2020年度) 特別  - その他