Research on transplantation of hepatic stem cells and development of experimental mouse models for hepatitis C virus

• Project/Area Number: 19590752
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: CHEN Cheng-hsin Tokyo Medical and Dental University, 医学部附属病院, 助教 (10376783)
• Co-Investigator(Kenkyū-buntansha): KAKINUMA Sei 東京大学, 医科学研究所, 特任助教・客員研究員 (30372444) ; SAKAMOTO Naoya 東京医科歯科大学, 医歯学総合研究科, 准教授 (10334418) ; WATANABE Mamoru 東京医科歯科大学, 医歯学総合研究科, 教授 (10175127)
• Co-Investigator(Renkei-kenkyūsha): SAKAMOTO Naoya 東京医科歯科大学, 医歯学総合研究科, 准教授 (10334418) ; WATANABE Mamoru 東京医科歯科大学, 医歯学総合研究科, 教授 (10175127)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2007: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 肝幹細胞 / C型肝炎ウイルス / 細胞移植 / FACS / shRNA / 肝細胞

Research Abstract

本研究において我々は、初代肝幹細胞を分離・培養し、これを利用して新規の細胞移植モデルをマウスにおいて確立すること、及びC型肝炎ウイルス(HCV)感染モデル動物の構築を行うことを目的として研究計画を遂行した。その結果、マウス肝幹細胞が高濃度に純化された画分を得る手法を確立し、その分化・増殖に制御する分子を数種類同定した。さらに、肝臓の約半分がドナー細胞によって置換される移植モデルの構築に成功した。そしてHCV感染動物に対する治療モデル系構築に成功し、研究計画を概ね達成した。

Research Products

• [Journal Article] Analyses of cell surface molecules on hepatic stem/progenitor cells in mouse fetal liver (2009)
  • Author(s): Kakinuma S, Ohta H, Kamiya A, Yamazaki Y, Oikawa T, Okada K, and Nakauchi H
  • Journal Title: J Hepatol (in press)
  • Peer Reviewed
• [Journal Article] Two flavonoidsextracts from Glycyrrhizae radix inhibit in vitrohepatitis C virus replication (2009)
  • Author(s): Sekine-Osajima Y, Sakamoto N, Nakagawa M, Itsui Y, Tasaka M, Nishimura-Sakurai Y, Chen CH, Suda G, Mishima K, Onuki Y, Yamamoto M, Maekawa S, Enomoto N, Kanai T, Tsuchiya K and Watanabe M
  • Journal Title: Hepatol Res 39 Pages: 60-9
  • Peer Reviewed
• [Journal Article] Signaling pathway via TNFa/NFkB in intestinal epithelial cells may be directly involved in colitis-associated carcinogenesis (2009)
  • Author(s): Onizawa M, Nagaishi T, Kanai T, Nagano KI, Oshima S, Nemoto Y, Yoshioka A, Totsuka T, Okamoto R, Nakamura T, Sakamoto N, TsuchiyaK, Aoki K, Ohya K, Yagita H and Watanabe M
  • Journal Title: Am J Physol Gastrntest Liver Physiol (Epub ahead of print)
  • Peer Reviewed
• [Journal Article] Sall4 regulates cell fate decision in fetal hepaticstem/progenitor cells (2009)
  • Author(s): Oikawa T, Kamiya A, Kakinuma S, Zeniya M, Nishinakamura R, Tajiri H and Nakauchi H
  • Journal Title: Gasroenterology 139 Pages: 1000-11
  • Peer Reviewed
• [Journal Article] Musashi-1 suppressesexpression of Paneth cell-specific genes inhuman intestinal epithelial cells (2009)
  • Author(s): Murayama M, Okamoto R, Tsuchiya K, Akiyama J, Nakamura T, Sakamoto N, Kanai T and Watanabe M
  • Journal Title: Gasroenterology 44 Pages: 173-82
  • Peer Reviewed
• [Journal Article] Hepatic stem/progenitor cells and stem-cell transplantation for the treatment of liver disease (2009)
  • Author(s): Kakinuma S, Nakauchi H and Watanabe M
  • Journal Title: J Gasroenterology 44 Pages: 167-72
  • NAID: 10025173601
• [Journal Article] Two flavonoids extracts from Glycyrrhizae radix inhibit in vitro hepatitis C virus replication. (2009)
  • Author(s): Sekine-Osajima Y, ChenCH., et al.
  • Journal Title: Hepatology Research 39 Pages: 60-69
  • NAID: 10025324061
  • Peer Reviewed
• [Journal Article] Sall4 regulates cell fate decision of fetal hepatic stem/progenitor cells (2009)
  • Author(s): Oikawa T, Kakinuma S, etal.
  • Journal Title: Gastroenterology 136 Pages: 1000-1011
  • Peer Reviewed
• [Journal Article] Hepatic stem/progenitor eelfor the treatment of liver ds and stem-cell transplantationsease. (2009)
  • Author(s): Kakinuma S, et al.
  • Journal Title: J Gastroenterol 44 Pages: 167-172
• [Journal Article] Immunosenescent colitogenicCD4(+) T cells convert to regulatory cells and Dsuppress colitis (2008)
  • Author(s): Totsuka T, Kanai T, Nemoto Y, Tomita T, Tsuchiya K, Sakamoto N, Okamoto R and Watanabe M
  • Journal Title: Eur J Immunol 38 Pages: 1275-86
  • Peer Reviewed
• [Journal Article] Systemic, but not intestinal, IL-7is essential for the persistence of chronic colitis (2008)
  • Author(s): Tomita T, Kanai T, Nemoto Y, Totsuka T, Okamoto R, Tsuchiya K, Sakamoto N and Watanabe M
  • Journal Title: J Immunol 180 Pages: 383-90
  • Peer Reviewed
• [Journal Article] Colitogenic CD4(+) effector-memory T cellsactively recirculate in chronic colitic mice (2008)
  • Author(s): Tomita T, Kanai T, Nemoto Y, Fujii T,Nozaki K, Okamoto R, Tsuchiya K, Nakamura T,Sakamoto N, Totsuka T and Watanabe M
  • Journal Title: In flamm Bowel Dis
  • Peer Reviewed
• [Journal Article] Continuous generation ofcolitogenic CD4(+) T cells in persistent colitis (2008)
  • Author(s): Tomita T, Kanai T, Fujii T, Nemoto Y,Okamoto R, Tsuchiya K, Totsuka T, Sakamoto N and Watanabe M
  • Journal Title: Eur J Immunol 38 Pages: 1264-74
  • Peer Reviewed
• [Journal Article] MyD88-dependentpathway in T cells directly modulates theexpansion of colitogenic CD4+ T cells in chroniccolitis (2008)
  • Author(s): Tomita T, Kanai T, Fujii T, Nemoto Y, Okamoto R, Tsuchiya K, Totsuka T, SakamotoN, Akira S and Watanabe M
  • Journal Title: J Immunol 180 Pages: 5291-9
  • Peer Reviewed
• [Journal Article] Development of plaque assays forhepatitis C virus-JFH1 strain and isolation ofmutants with enhanced cytopathogenicity and replication capacity (2008)
  • Author(s): Sekine-Osajima Y, Sakamoto N, MishimaK, Nakagawa M, Itsui Y, Tasaka M,Nishimura-Sakurai Y, Chen CH, Kanai T, Tsuchiya K, Wakita T, Enomoto N and Watanabe M
  • Journal Title: Virology 371 Pages: 71-85
  • Peer Reviewed
• [Journal Article] Negative feedback regulation of colitogenicCD4(+) T cells by increased granulopoiesis (2008)
  • Author(s): Nemoto Y, Kanai T, Tohda S, Totsuka T,Okamoto R, Tsuchiya K, Nakamura T, SakamotoN, Fukuda T, Miura O, Yagita H and Watanabe M
  • Journal Title: In flamm Bowel Dis
  • Peer Reviewed
• [Journal Article] Effects of immunosuppressantson the progression of hepatitis C in hepatitis Cvirus-positive renal transplantation and theusefulness of interferon therapy (2008)
  • Author(s): Nanmoku K, Imaizumi R, Tojimbara T, Nakajima I, Fuchinoue S, Sakamoto N, Watanabe M and Teraoka S
  • Journal Title: Transplant Proc 40 Pages: 2382-5
  • Peer Reviewed
• [Journal Article] Prospero-relatedhomeobox 1 and liver receptor homolog 1coordinately regulate long-term proliferation ofmurine fetal hepatoblasts (2008)
  • Author(s): Kamiya A, Kakinuma S, Onodera M, Miyajima A and Nakauchi H
  • Journal Title: Hepatology 48 Pages: 252-64
  • Peer Reviewed
• [Journal Article] Blockade of NKG2D signaling prevents the development ofmurine CD4+ T cell-mediated colitis (2008)
  • Author(s): Ito Y, Kanai T, Totsuka T, Okamoto R, Tsuchiya K, Nemoto Y, Yoshioka A, Tomita T, Nagaishi T, Sakamoto N, Sakanishi T, OkumuraK, Yagita H and Watanabe M
  • Journal Title: Am J Physiol Gastro intest Liver Physiol 294
  • Peer Reviewed
• [Journal Article] FTY720 suppresses the development of colitis in lymphoid-null mice by modulating the trafficking of colitogenic CD4+ T cells in bone marrow (2008)
  • Author(s): Fujii T, Tomita T, Kanai T, Nemoto Y,Totsuka T, Sakamoto N, Nakamura T, TsuchiyaK, Okamoto R and Watanabe M
  • Journal Title: EurJ Immunol 38 Pages: 3290-303
  • Peer Reviewed
• [Journal Article] Proteasomal degradation ofAtoh1 by aberrant Wnt signaling maintains theundifferentiated state of colon cancer (2008)
  • Author(s): Aragaki M, Tsuchiya K, Okamoto R,Yoshioka S, Nakamura T, Sakamoto N, Kanai T and Watanabe M
  • Journal Title: Biochem Biophys Res Commun 368 Pages: 923-9
  • Peer Reviewed
• [Journal Article] TCF/LEF経路を介した肝幹・前駆細胞成熟化の調節機構 (2008)
  • Author(s): 柿沼晴
  • Journal Title: 臨床消化器内科 23巻13号 Pages: 1813-1819
• [Journal Article] Prospero-related homeobox 1 and liver receptor homolog 1 coordinately regulate long-term proliferation of murine fetal hepatoblasts. (2008)
  • Author(s): Kamiya A, Kakinuma s, etal.
  • Journal Title: Hepatology 48 Pages: 252-264
  • Peer Reviewed
• [Journal Article] Development of plaque assays for hepatitis C virus-JFH1 strain and isolation of mutants with enhanced cytopathogenicity and replication capacity. (2008)
  • Author(s): Sekine-Osajima Y, Sakamoto N, Chen CH, Watanabe M, et. al.
  • Journal Title: Virology 371 Pages: 71-85
  • Peer Reviewed
• [Journal Article] Prospero-related homeobox 1 and liver receptor homolog 1 coordinately regulate long-term proliferation of murine fetal hepatoblasts. (2008)
  • Author(s): Kamiya A, Kakinuma S, et. al.
  • Journal Title: Hepatology (印刷中)
  • Peer Reviewed
• [Journal Article] Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA. (2008)
  • Author(s): Sakamoto N, Chen CH, Watanabe M, et. al.
  • Journal Title: Journal of Gastroenterology and Hepatology (印刷中)
  • Peer Reviewed
• [Journal Article] Hepatitis C virus non-structural proteins responsible for suppression of the RIG-I/Cardif-induced interferon response (2007)
  • Author(s): Tasaka M, Sakamoto N, Itakura Y,Nakagawa M, Itsui Y, Sekine-Osajima Y,Nishimura-Sakurai Y, Chen CH, Yoneyama M,Fujita T, Wakita T, Maekawa S, Enomoto N and Watanabe M
  • Journal Title: J Gen Virol 88 Pages: 3323-33
  • Peer Reviewed
• [Journal Article] Bone morphogenetic protein-7 and interferon-alpha synergistically suppresshepatitis C virus replicon (2007)
  • Author(s): Sakamoto N, Yoshimura M, Kimura T, Toyama K, Sekine-Osajima Y, Watanabe M and Muramatsu M
  • Journal Title: Biochem Biophys Res Commun 357 Pages: 467-73
  • Peer Reviewed
• [Journal Article] Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA (2007)
  • Author(s): Sakamoto N, Tanabe Y, Yokota T, Satoh K, Sekine-Osajima Y, Nakagawa M, Itsui Y, Tasaka M, Sakurai Y, Chen CH, Yano M, Ohkoshi S,Aoyagi Y, Maekawa S, Enomoto N, Kohara M and Watanabe M
  • Journal Title: J Gastroenterol Hepatol
  • Peer Reviewed
• [Journal Article] Hepatitis C virus non-structural proteins responsible for suppression of the RIG-I/Cardif-induced interferon response. (2007)
  • Author(s): Tasaka M, Sakamoto N, Chen CH, Watanabe M, et. al.
  • Journal Title: Journal of General Virology 88 Pages: 3323-3333
  • Peer Reviewed
• [Presentation] 肝幹/前駆細胞表面抗原の網羅的解析と細胞移植 (2008)
  • Author(s): 柿沼晴、紙谷聡英、中内啓光
  • Organizer: JDDW2008(第12回日本肝臓学会大会ワークショップ/肝再生 ; 基礎と臨床のBridge研究)
  • Place of Presentation: 東京
  • Year and Date: 2008-10-18
• [Presentation] Analysis of cell surface proteins on hepatic stem/progenitorcells in fetal liver (2008)
  • Author(s): Kakinuma s, et al.
  • Organizer: FASEB summer research conference
  • Place of Presentation: Snowmass CO USA
  • Year and Date: 2008-08-04
• [Presentation] 胎生中期肝幹/前駆細胞表面抗原の解析と細胞移植 (2008)
  • Author(s): 柿沼晴、太田春彦、紙谷聡英、中内啓光
  • Organizer: 第15回肝細胞研究会(シンポジウム/肝障害と幹細胞~肝修復における幹細胞の役割)
  • Place of Presentation: 静岡
  • Year and Date: 2008-06-28
• [Presentation] 3cmを超える肝細胞癌に対する当科の治療方法と成績 (2008)
  • Author(s): 陳正新、坂本直哉、井津井康浩、中川美奈、櫻井幸、須田剛生、山本満千、小貫優子、三島果子、渡辺守
  • Organizer: 第44回日本肝臓学会総会
  • Place of Presentation: 松山
  • Year and Date: 2008-06-06
• [Presentation] 肝幹/前駆細胞表面抗原のFACSによる網羅的解析 (2008)
  • Author(s): 柿沼晴、太田春彦、紙谷聡英、中内啓光
  • Organizer: 第44回日本肝臓学会総会
  • Place of Presentation: 松山(口演)
  • Year and Date: 2008-06-06
• [Presentation] 3cmを超える肝細胞癌に対する当科の治療方法と成績 (2008)
  • Author(s): 陳 正新, 他
  • Organizer: 第4 4回日本肝臓学会総会
  • Place of Presentation: 愛媛県県民文化会蔀
  • Year and Date: 2008-06-05
• [Presentation] Flowcytometric analysis of cell surfacemolecules on mouse fetal hepaticstem/progenitor cells (2008)
  • Author(s): Kakinuma S, Kamiya A, Ohta H, Nakauchi H
  • Organizer: FASEB summer research conference , Liver growth,development and disease
  • Place of Presentation: Snowmass, CO, USA
• [Presentation] ラジオ波熱凝固療法における第二世代超音波造影剤の有用性 (2007)
  • Author(s): 陳正新、坂本直哉、井津井康浩、中川美奈、筬島裕子、田坂めぐみ、櫻井幸、渡辺守
  • Organizer: JDDW2007(第10回日本肝臓学会大会)
  • Place of Presentation: 神戸
  • Year and Date: 2007-10-18
• [Presentation] TCF/LEF活性化による肝幹/前駆細胞の分化・増殖の調節 (2007)
  • Author(s): 柿沼晴、紙谷聡英、中内啓光
  • Organizer: JDDW2007(第10回日本肝臓学会大会ワークショップ/肝の再生医学・医療/人工肝臓の現状と展望)
  • Place of Presentation: 神戸
  • Year and Date: 2007-10-18
• [Presentation] TCF/LEF活性化による肝幹/前駆細胞の分化・増殖の調節 (2007)
  • Author(s): 柿沼 晴、他
  • Organizer: 第11回日本肝臓学会大会ワークショップ
  • Place of Presentation: 神戸
  • Year and Date: 2007-10-18
• [Presentation] 肝幹/前駆細胞におけるTCF/LEF活性化 (2007)
  • Author(s): 柿沼晴、紙谷聡英、中内啓光
  • Organizer: 第14回肝細胞研究会(ワークショップ/肝幹細胞研究の現況と展望)
  • Place of Presentation: 鹿児島
  • Year and Date: 2007-06-23
• [Presentation] Wnt/TCF signaling pathwayを介した肝前駆細胞成熟化の調節機構 (2007)
  • Author(s): 柿沼晴、紙谷聡英、中内啓光
  • Organizer: 第43回日本肝臓学会総会(ワークショップ/肝再生とその臨床応用)
  • Place of Presentation: 東京
  • Year and Date: 2007-05-31
• [Presentation] 当科における高齢肝細胞癌患者に対するラジオ波熱凝固療法の治療成績 (2007)
  • Author(s): 陳正新、坂本直哉、渡辺守
  • Organizer: 第43回日本肝臓学会総会
  • Place of Presentation: 東京
  • Year and Date: 2007-05-31
• [Book] 幹細胞を用いた肝再生医療の可能性 (2009)
  • Author(s): 柿沼晴
  • Publisher: iPS細胞の産業的応用技術(シーエムシー出版)(印刷中)
• [Book] Wnt-TCF/LEF経路による肝幹細胞の分化・増殖調節機構の解析 (2008)
  • Author(s): 柿沼晴
  • Publisher: Liver Forum in Kyoto第10回学術集会記録集(メディカルトリビューン)
• [Book] 幹細胞-肝幹細胞を用いた細胞移植の可能性 (2007)
  • Author(s): 柿沼晴
  • Publisher: 再生医療技術の最前線(シーエムシー出版)
• [Book] 再生医療技術の最前線:幹細胞-肝幹細胞を用いた細胞移植の可能性 (2007)
  • Author(s): 柿沼 晴(岡野光夫他監修)分著
  • Publisher: シーエムシー出版