Faculty Profiles - KAGAWA Shunsuke

Papers - KAGAWA Shunsuke

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Oncolytic virotherapy for osteosarcoma

Pharm stage 16 ( 12 ) 54 - 59 2017

HER2陽性胃癌に対する新たな治療戦略

金谷信彦, 黒田新士, 久保田哲史, 森廣俊昭, 菊地覚次, 西崎正彦, 田澤大, 香川俊輔, 藤原俊義

癌と化学療法 44 ( 10 ) 883 - 885 2017

超高齢者の進行食道癌に対し積極的に集学的治療を行い長期無再発生存中の1例

松三雄騎, 白川靖博, 田邊俊介, 河本慧, 前田直見, 二宮卓之, 野間和広, 西崎正彦, 香川俊輔, 藤原俊義

癌と化学療法 44 ( 12 ) 1784 - 1786 2017

胃癌術後再発病変に対する複数回の放射線療法が奏功し長期生存が得られた1例

堀直人, 香川俊輔, 菊地覚次, 黒田新士, 渡邉めぐみ, 坂本修一, 香川哲也, 桒田和也, 久保田哲史, 岸本浩行, 西﨑正彦, 片山敬久, 藤原俊義

癌と化学療法 44 ( 2 ) 165 - 167 2017

Language：Japanese Publisher：(株)癌と化学療法社

胃癌術後再発に対しての第一選択は化学療法であるが、胃癌術後複数箇所のリンパ節転移に対して、3回の放射線療法(radio therapy:RT)により長期の病勢制御が可能であった症例を経験したので報告する。症例は70歳、男性。胃癌に対して幽門側胃切除を行い、Stage IIICであった。S-1の補助化学療法を行ったが、3年経過後に膵頭後部リンパ節再発を来した。他に再発病巣なくRTを行い、奏効が得られた。その後、化学療法中に出現した腹部大動脈周囲リンパ節、縦隔リンパ節再発に対してもそれぞれRTを施行し、再発から4年7ヵ月間、化学療法抵抗性であった再発病巣は良好に制御されている。胃癌術後再発におけるRTは、その多くが多発病巣を呈するため適応にはなりにくく、胃周辺臓器への有害事象も考慮する必要があるが、本症例のような化学療法抵抗性の単発病変再発の局所制御には有効性を発揮することがあり、治療の選択肢として重要である。(著者抄録)

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Cell-cycle-dependent drug-resistant quiescent cancer cells induce tumor angiogenesis after chemotherapy as visualized by real-time FUCCI imaging

Shuya Yano, Kiyoto Takehara, Hiroshi Tazawa, Hiroyuki Kishimoto, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara, Robert M. Hoffman

CELL CYCLE 16 ( 5 ) 406 - 414 2017

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

We previously demonstrated that quiescent cancer cells in a tumor are resistant to conventional chemotherapy as visualized with a fluorescence ubiquitination cell cycle indicator (FUCCI). We also showed that proliferating cancer cells exist in a tumor only near nascent vessels or on the tumor surface as visualized with FUCCI and green fluorescent protein (GFP)-expressing tumor vessels. In the present study, we show the relationship between cell-cycle phase and chemotherapy-induced tumor angiogenesis using in vivo FUCCI real-time imaging of the cell cycle and nestin-driven GFP to detect nascent blood vessels. We observed that chemotherapy-treated tumors, consisting of mostly of quiescent cancer cells after treatment, had much more and deeper tumor vessels than untreated tumors. These newly-vascularized cancer cells regrew rapidly after chemotherapy. In contrast, formerly quiescent cancer cells decoyed to S/G(2) phase by a telomerase-dependent adenovirus did not induce tumor angiogenesis. The present results further demonstrate the importance of the cancer-cell position in the cell cycle in order that chemotherapy be effective and not have the opposite effect of stimulating tumor angiogenesis and progression.

DOI： 10.1080/15384101.2016.1220461

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食道がんに対する放射線併用ウイルス療法の臨床研究

藤原俊義, 田澤大, 田邊俊介, 香川俊輔, 白川靖博

日本臨床 75 ( 5 ) 746 - 751 2017

HOZOT細胞を用いた腫瘍融解ウイルス製剤のDDS

田澤大, 大西哲平, 香川俊輔, 中村修治, 藤原俊義

BIO INDUSTRY 34 ( 4 ) 16 - 23 2017

噴門側胃切除術後再建：手縫い腹腔鏡下体腔内手縫い「観音開き法」食道残胃吻合

西﨑正彦, 黒田新士, 菊地覚次, 垣内慶彦, 松村年久, 渡邉めぐみ, 香川俊輔, 藤原俊義

臨床外科 72 ( 4 ) 412 - 415 2017

観音開き法を用いた噴門機能温存

西﨑正彦, 黒田新士, 菊地覚次, 垣内慶彦, 香川俊輔, 藤原俊義

手術 71 ( 1 ) 21 - 25 2017

In Vivo Selection of Intermediately- and Highly-Malignant Variants of Triple-negative Breast Cancer in Orthotopic Nude Mouse Models

Shuya Yano, Kiyoto Takehara, Hiroyuki Kishimoto, Hiroshi Tazawa, Yasuo Urata, Shunsuke Kagawa, Michael Bouvet, Toshiyoshi Fujiwara, Robert M. Hoffman

ANTICANCER RESEARCH 36 ( 12 ) 6273 - 6277 2016.12

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

Aim: Triple-negative breast cancer (TNBC), defined by the absence of receptors for estrogen, progesterone and human epithelial receptor 2 (HER2), is a recalcitrant disease in need of effective therapy. We previously isolated very-highly metastatic variants of the TNBC cell line MDA-MB-231 using serial orthotopic implantation of MDA-MB-231 human breast cancer cells in nude mice. Materials and Methods: MDA-MB-231 cells expressing red fluorescent protein (MDA-MB-231-RFP) (1x10(7) cells/site) were initially injected subcutaneously in the flank of nude mice. After the subcutaneous tumors grew, they were harvested and cut into small pieces for orthotopic implantation into the right lower mammary gland. After the orthotopic tumors grew, they were resected and cut into small pieces and orthotopically re-implanted into the mammary gland of nude mice. The tumors grew and metastasized to lymph nodes. The lymph node metastases were harvested and cut into small pieces and orthotopically re-implanted into the mammary gland of nude mice. After the orthotopic tumors grew, the tumor was removed leaving residual cancer cells, which grew and metastasized to lymph nodes. The lymph node metastases were harvested, cut into pieces and orthotopically re-implanted into the mammary gland of nude mice for three or seven cycles in order to isolate intermediately, or highly-metastatic variants, respectively. Results: The degree of malignancy of isolated variants of MDA-MB-231 depends on the extent of orthotopic transplantation. Serial transplantation resulted in MDA-MB-231-RFP which significantly produced larger tumors compared with the parental MDA-MB-231-RFP. Serial orthotopic implantation for three cycles resulted in intermediately-metastatic variants of MDA-MB-231-RFP. MDA-MB-231-RFP serially orthotopically transplanted seven times significantly metastasized more to lymph nodes compared with parental MDA-MB-231-RFP cells and the intermediately-metastatic variant. The highly-metastatic variant MDA-MB-231-RFP cells significantly metastasized to the lung to a greater extent compared with parental MDA-MB-231-RFP and intermediate variants of MDA-MB-231-RFP. Conclusion: These results suggest that the number of serial orthotopic transplantations of MDA-MB-231-RFP correlates positively with tumor aggressiveness, and the intermediately- and highly-metastatic variants can serve as models of metastatic progression of TNBC.

DOI： 10.21873/anticanres.11222

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Tumor-targeting adenovirus OBP-401 inhibits primary and metastatic tumor growth of triple-negative breast cancer in orthotopic nude-mouse models

Shuya Yano, Kiyoto Takehara, Hiroyuki Kishimoto, Hiroshi Tazawa, Yasuo Urata, Shunsuke Kagawa, Michael Bouvet, Toshiyoshi Fujiwara, Robert M. Hoffman

ONCOTARGET 7 ( 51 ) 85273 - 85282 2016.12

Language：English Publishing type：Research paper (scientific journal) Publisher：IMPACT JOURNALS LLC

Our laboratory previously developed a highly-invasive, triple-negative breast cancer (TNBC) variant using serial orthotopic implantation of the human MDA-MB-231 cell line in nude mice. The isolated variant was highly-invasive in the mammary gland and lymphatic channels and metastasized to lymph nodes in 10 of 12 mice compared to 2 of 12 of the parental cell line. In the present study, the tumor-selective telomerase dependent OBP-401 adenovirus was injected intratumorally (i. t.) (1 x 10(8) PFU) when the high-metastatic MDA-MB-231 primary tumor expressing red fluorescent protein (MDA-MB-231-RFP) reached approximately 500 mm3 (diameter; 10 mm). The mockinfected orthotopic primary tumor grew rapidly. After i. t. OBP-401 injection, the growth of the orthotopic tumors was arrested. Six weeks after implantation, the fluorescent area and fluorescence intensity showed no increase from the beginning of treatment. OBP-401 was then injected into high-metastatic MDA-MB-231-RFP primary orthotopic tumor growing in mice which already had developed metastasis within lymphatic ducts. All 7 of 7 control mice subsequently developed lymph node metastasis. In contrast, none of 7 mice which received OBP-401 had lymph node metastasis. Seven of 7 control mice also had gross lung metastasis. In contrast, none of the 7 mice which received OBP-401 had gross lung metastasis. Confocal laser microscopy imaging demonstrated that all control mice had diffuse lung metastases. In contrast, all 7 mice which received OBP-401 only had a few metastatic cells in the lung. OBP-401 treatment significantly extended survival of the treated mice.

DOI： 10.18632/oncotarget.13296

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[A Case of Multiple Liver Metastasis from Gastric Cancer Successfully Treated with CPT-11 as Third-Line Chemotherapy].

Satoru Kikuchi, Masahiko Nishizaki, Shinji Kuroda, Shunsuke Kagawa, Toshiyoshi Fujiwara

Gan to kagaku ryoho. Cancer & chemotherapy 43 ( 12 ) 2219 - 2221 2016.11

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

A 63-year-old man underwent proximal gastrectomy for early gastric cancer(pT1bN0M0, pStage I A). Twenty months after surgery, abdominal CT scans revealed multiple liver metastases. S-1 plus CDDP therapy was administered as first-line chemotherapy. After treatment, CT scans revealed tumor progression and nab-PTX was administrated. This treatment was ineffective; therefore, CPT-11 was administrated as third-line chemotherapy. Treatment with CPT-11 resulted in marked tumor reduction and improved the QOL of the patient; partial response was maintained for 8 months. After 17 courses of CPT-11 treatment, tumor regrowth was detected, and the patient was treated with S-1 plus oxaliplatin, DTX, and ramucirumab. Subsequently, the patient died of cancer 31 months after tumor recurrence. CPT-11 is potentially a key drug for the prevention of liver metastasis of gastric cancer, and using all active agents in patients with advanced gastric cancer over several lines of therapy could prolong survival.

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Real-time in-vivo image-guided cell-cycle perturbation to increase tumor chemosensitivity

Shuya Yano, Hiroshi Tazawa, Hiroyuki Kishimoto, Shunsuke Kagawa, Yoshihiko Kadowaki, Nobuhiro Ishido, Takahiro Okamoto, Yasuo Urata, Kiyoto Takehara, Robert M. Hoffman, Toshiyoshi Fujiwara

MOLECULAR CANCER RESEARCH 14 2016.11

Fluorescence-guided surgery of a highly-metastatic variant of human triple-negative breast cancer targeted with a cancer-specific GFP adenovirus prevents recurrence

Shuya Yano, Kiyoto Takehara, Shinji Miwa, Hiroyuki Kishimoto, Hiroshi Tazawa, Yasuo Urata, Shunsuke Kagawa, Michael Bouvet, Toshiyoshi Fujiwara, Robert M. Hoffman

ONCOTARGET 7 ( 46 ) 75635 - 75647 2016.11

Language：English Publishing type：Research paper (scientific journal) Publisher：IMPACT JOURNALS LLC

We have previously developed a genetically-engineered GFP-expressing telomerase-dependent adenovirus, OBP-401, which can selectively illuminate cancer cells. In the present report, we demonstrate that targeting a triple-negative highinvasive human breast cancer, orthotopically-growing in nude mice, with OBP-401 enables curative fluorescence-guided surgery (FGS). OBP-401 enabled complete resection and prevented local recurrence and greatly inhibited lymph-node metastasis due to the ability of the virus to selectively label and subsequently kill cancer cells. In contrast, residual breast cancer cells become more aggressive after bright (white)light surgery (BLS). OBP-401-based FGS also improved the overall survival compared with conventional BLS. Thus, metastasis from a highly-aggressive triple-negative breast cancer can be prevented by FGS in a clinically-relevant mouse model.

DOI： 10.18632/oncotarget.12314

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Tumor-specific delivery of biologics by a novel T-cell line HOZOT

Teppei Onishi, Hiroshi Tazawa, Yuuri Hashimoto, Makoto Takeuchi, Takeshi Otani, Shuji Nakamura, Fuminori Sakurai, Hiroyuki Mizuguchi, Hiroyuki Kishimoto, Yuzo Umeda, Yasuhiro Shirakawa, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara

Scientific Reports 6 2016.11

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

"Cell-in-cell" denotes an invasive phenotype in which one cell actively internalizes in another. The novel human T-cell line HOZOT, established from human umbilical cord blood, was shown to penetrate a variety of human cancer cells but not normal cells. Oncolytic viruses are emerging as biological therapies for human cancers; however, efficient viral delivery is limited by a lack of tumor-specific homing and presence of pre-existing or therapy-induced neutralizing antibodies. Here, we report a new, intriguing approach using HOZOT cells to transmit biologics such as oncolytic viruses into human cancer cells by cell-in-cell invasion. HOZOT cells were successfully loaded via human CD46 antigen with an attenuated adenovirus containing the fiber protein of adenovirus serotype 35 (OBP-401/F35), in which the telomerase promoter regulates viral replication. OBP-401/F35-loaded HOZOT cells were efficiently internalized into human cancer cells and exhibited tumor-specific killing by release of viruses, even in the presence of anti-viral neutralizing antibodies. Moreover, intraperitoneal administration of HOZOT cells loaded with OBP-401/F35 significantly suppressed peritoneally disseminated tumor growth in mice. This unique cell-in-cell property provides a platform for selective delivery of biologics into human cancer cells, which has important implications for the treatment of human cancers.

DOI： 10.1038/srep38060

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高リスク神経芽腫に対するhTERT標的化膿瘍融解ウイルス療法(hTERT-targeted oncolytic virotherapy against high-risk neuroblastomas)

谷本光隆, 田澤大, 家田偉史, 納所洋, 尾山貴徳, 浦田泰生, 香川俊輔, 野田卓男, 藤原俊義

日本小児血液・がん学会雑誌 53 ( 4 ) 202 - 202 2016.11

Novel HER2-Targeted therapy combined with gold nanoparticles

Tetsushi Kubota, Shinji Kuroda, Toshiaki Morihiro, Hiroshi Tazawa, Shunsuke Kagawa, Toshiyoshi Fujiwara

Japanese Journal of Cancer and Chemotherapy 43 ( 10 ) 1237 - 1239 2016.10

Language：Japanese Publisher：Japanese Journal of Cancer and Chemotherapy Publishers Inc.

Trastuzumab (Tmab) is a humanized monoclonal antibody that binds to the human epidermal growth factor receptor 2 (HER2). It is clinically used for HER2-positive breast and gastric cancers
however, the use of Tmab is restricted to tumors expressing high levels of HER2 (accounting for only 20% of tumors), and Tmab cannot be used for tumors resistant to Tmab. Although novel HER2-Targeted agents have been developed to treat Tmab-resistant tumors, none of these have shown clinical efficacy in gastric cancer patients. Recent developments in nanotechnology have had a significant impact on the field of medicine. Gold nanoparticles (AuNPs), which show characteristics such as in vivo stability and ease of surface functionali- zation, have been developed as therapeutic and contrast agents for medical applications. Previous studies show that AuNPs exert cytotoxic effects through autophagy and apoptosis
therefore, AuNPs in combination with tumor-Targeting antibodies are attractive therapeutic agents. In this study, we developed HER2-Targeted AuNPs (Tmab-AuNPs) and showed that they had a potent antitumor effect on Tmab-resistant cell lines. In addition, Tmab-AuNPs were effective against HER2-negative gastric cancer cell lines when HER2 was artificially overexpressed. Thus, our results indicate that Tmab-AuNPs may overcome the shortcomings of Tmab-based therapy.

Scopus

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金ナノ粒子を用いた新規HER2標的抗体療法

久保田哲史, 黒田新士, 森廣俊昭, 田澤大, 香川俊輔, 藤原俊義

癌と化学療法 43 ( 10 ) 1237 - 1239 2016.10

Language：Japanese Publisher：(株)癌と化学療法社

trastuzumab(Tmab)は、human epidermal growth factor receptor 2(HER2)に対するヒト化モノクローナル抗体で、現在乳癌や胃癌に対して臨床使用され予後を改善しているが、HER2の低発現(約20%)やTmab抵抗性の問題がある。Tmab抵抗例に対して新たなHER2標的製剤が開発されているが胃癌に対して臨床的に有効なものはまだなく、新たな治療法が望まれる。ナノ技術はその発展に伴い医療へ応用されてきている。金ナノ粒子は生体内での安定性と表面修飾の容易性などの特性があり、autophagyやoxidative stressを介したアポトーシスを誘導することも報告されている。われわれは、HER2標的金ナノ粒子製剤(Tmab-AuNPs)を作製し、Tmab抵抗性胃癌細胞株に対する抗腫瘍効果とHER2細胞外領域を強制発現させたHER2陰性株に対する抗腫瘍効果を確認しており、Tmab-AuNPsがTmab抵抗性胃癌に対する新たなHER2標的製剤になり得ると考えている。(著者抄録)

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A Phase II Clinical Trial of the Efficacy and Safety of Short-term (3 days) Enoxaparin for the Prevention of Venous Thromboembolism after Gastric Cancer Surgery

Shinji Kuroda, Satoru Kikuchi, Masahiko Nishizaki, Shunsuke Kagawa, Shiro Hinotsu, Toshiyoshi Fujiwara

ACTA MEDICA OKAYAMA 70 ( 5 ) 401 - 404 2016.10

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

Although intermittent pneumatic compression (IPC) has become common as perioperative prophylaxis for venous thromboembolism (VTE) consisting of pulmonary thromboembolism (PE) and deep vein thrombosis (DVT), the prophylactic effect against VTE, especially lethal PE, is not yet satisfactory. Therefore, pharmacologic prophylaxis, such as with enoxaparin, is desirable. While the efficacy and safety of enoxaparin have been proven in several clinical trials, concern about bleeding with long-term (at least 7 days) use have potentially decreased its widespread adoption. We have launched a phase II study to evaluate the efficacy and safety of short-term (3 days) enoxaparin, in which a total of 70 gastric cancer patients undergoing gastrectomy will be recruited, and the primary endpoint is the incidence of DVT. This study could contribute to making pharmacologic prophylaxis for VTE more common.

DOI： 10.18926/AMO/54601

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Efficacy of a Cell-Cycle Decoying Killer Adenovirus on 3-D Gelfoam (R)-Histoculture and Tumor-Sphere Models of Chemo-Resistant Stomach Carcinomatosis Visualized by FUCCI Imaging

Shuya Yano, Kiyoto Takehara, Hiroshi Tazawa, Hiroyuki Kishimoto, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara, Robert M. Hoffman

PLOS ONE 11 ( 9 ) doi: 10.1371/journal.pone.0162991 2016.9

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

Stomach cancer carcinomatosis peritonitis (SCCP) is a recalcitrant disease. The goal of the present study was to establish an in vitro-in vivo-like imageable model of SCCP to develop cell-cycle-based therapeutics of SCCP. We established 3-D Gelfoam (R) histoculture and tumor-sphere models of SCCP. FUCCI-expressing MKN-45 stomach cancer cells were transferred to express the fluorescence ubiquinized cell-cycle indicator (FUCCI). FUCCI-expressing MKN-45 cells formed spheres on agarose or on Gelfoam (R) grew into tumor-like structures with G(0)/G(1) cancer cells in the center and S/G(2) cancer cells located in the surface as indicated by FUCCI imaging when the cells fluoresced red or green, respectively. We treated FUCCI-expressing cancer cells forming SCCP tumors in Gelfoam (R) histoculture with OBP-301, cisplatinum (CDDP), or paclitaxel. CDDP or paclitaxel killed only cycling cancer cells and were ineffective against G1/G2 MKN-45 cells in tumors growing on Gelfoam (R). In contrast, the telomerase-dependent adenovirus OBP-301 decoyed the MKN45 cells in tumors on Gelfoam (R) to cycle from G(0)/G(1) phase to S/G(2) phase and reduced their viability. CDDP-or paclitaxel-treated MKN-45 tumors remained quiescent and did not change in size. In contrast, OB-301 reduced the size of the MKN-45 tumors on Gelfoam 1. We examined the cell cycle-related proteins using Western blotting. CDDP increased the expression of p53 and p21 indicating cell cycle arrest. In contrast, OBP-301 decreased the expression of p53 and p21 Furthermore, OBP-301 increased the expression of E2F and pAkt as further indication of cell cycle decoy. This 3-D Gelfoam (R) histoculture and FUCCI imaging are powerful tools to discover effective therapy of SCCP such as OBP-301.

DOI： 10.1371/journal.pone.0162991

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