Ｃ型肝炎ウイルスの持続感染化、生体防御応答による慢性炎症・肝がん発症機序の解明

• Project Area: Conversion of tumor-regulation vector to intercept oncogenic spiral accelerated by infection and inflammation
• Project/Area Number: 25114712
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• Principal Investigator: 小原 道法 公益財団法人東京都医学総合研究所, ゲノム医科学研究分野, 特任研究員 (10250218)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥12,220,000 (Direct Cost: ¥9,400,000、Indirect Cost: ¥2,820,000); Fiscal Year 2014: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2013: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
• Keywords: C型肝炎ウイルス / 持続感染 / 肝線維化 / スイッチングトランスジェニックマウス / Wnt/beta-cateninシグナル阻害 / 感染症 / 肝炎 / 発がん / ウイルス / 動物モデル

Outline of Annual Research Achievements

C型肝炎ウイルス（HCV）は容易に持続感染化し、慢性肝炎から肝線維症、肝硬変を経て高率に肝細胞がんへと進展する。本邦には20万人ものC型肝硬変患者がいると推定され、国内肝硬変患者数の約70%に相当する。高度な肝線維症である肝硬変の治療は困難であり、発症原因であるHCV持続感染の根本治療を目的とした抗ウイルス剤の効果も十分ではない。肝線維症を含む肝硬変を改善する薬剤を開発するためには、免疫系が複雑に寄与する持続炎症による肝線維症から離脱できる方法を見出すことが重要である。そこで我々は、C型肝硬変における線維芽細胞活性化因子TGF-βの下流で活性化されるWnt/β-catenin/CBPシグナルの選択的阻害剤PRI-724に着目し、HCV誘発性肝線維症改善作用と作用機序の解析を行った。選択的Wnt/β-catenin/CBPシグナル阻害剤PRI-724は、HCV遺伝子型1bをスイッチング発現する肝線維症モデルマウスの肝臓中コラーゲン量およびコラーゲン線維を減少させた。薬剤投与後の血清中ALT値上昇は認められず、PRI-724の肝線維症改善作用は細胞死を伴わないことが示された。肝臓中mRNA発現解析により、HCV持続発現により抑制されたマトリックスメタロプロテアーゼ（MMP-8とMMP-9）の発現がPRI-724投与により誘導されることが明らかとなった。また、これらの主な産生細胞である好中球やマクロファージ数もPRI-724投与により増加し、肝線維症改善作用に寄与することが示唆された。PRI-724は、C型肝硬変に対する治療薬候補となると考えられた

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Inhibitory Effects of Pycnogenol on Hepatitis C Virus Replication. (2015)
  • Author(s): Ezzikouri S, Nishimura T, Kohara M, Benjelloun S, Kino Y, Inoue K, Matsumori A, Tsukiyama-Kohara K
  • Journal Title: Antiviral Research Volume: 113C Pages: 93-102
  • DOI: 10.1016/j.antiviral.2014.10.017
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The RNA Sensor RIG-I Dually Functions as an Innate Sensor and Direct Antiviral Factor for Hepatitis B Virus (2015)
  • Author(s): Sato S, Li K, Kameyama T, Hayashi T, Ishida Y, Murakami S, Watanabe T, Iijima S, Sakurai Y, Watashi K, Tsutsumi S, Sato Y, Akita H, Wakita T, Rice CM, Harashima H, Kohara M, Tanaka Y, Takaoka A
  • Journal Title: Immunity Volume: 42 Issue: 1 Pages: 123-132
  • DOI: 10.1016/j.immuni.2014.12.016
  • Peer Reviewed / Open Access
• [Journal Article] Recent insights into hepatitis B virus-host interactions. (2014)
  • Author(s): Ezzikouri S, Ozawa M, Kohara M, Elmdaghri N, Benjelloun S, Tsukivama-Kohara K.
  • Journal Title: Journal of Medical Virology Volume: 86 Issue: 6 Pages: 925-32
  • DOI: 10.1002/jmv.23916
  • Peer Reviewed / Open Access
• [Journal Article] B-cell-intrinsic hepatitis C virus expression leads to B-cell-lymphomagenesis and induction of NF-κB signalling (2014)
  • Author(s): Kasama Y, Mizukami T, Kusunoki H, Peveling-Oberhag J, Nishito Y, Ozawa M, Kohara M, Mizuochi T, Tsukiyama-Kohara K
  • Journal Title: PLoS One Volume: 9(3) Issue: 3 Pages: 91373-91373
  • DOI: 10.1371/journal.pone.0091373
  • Peer Reviewed / Open Access
• [Journal Article] In vivo therapeutic potential of Dicer-hunting siRNAs targeting infectious hepatitis C virus (2014)
  • Author(s): Watanabe T, Hatakeyama H, Matsuda-Yasui C, Sato Y, Sudoh M, Takagi A, Hirata Y, Ohtsuki T, Arai M, Inoue K, Harashima H, Kohara M
  • Journal Title: Scientific Reports Volume: in press Issue: 1 Pages: 4750-4750
  • DOI: 10.1038/srep04750
  • Peer Reviewed / Open Access
• [Journal Article] Resistance to cyclosporin A derives from mutations in hepatitis C virus nonstructural proteins. (2014)
  • Author(s): Arai M, Tsukiyama-Kohara K, Takagi A, Tobita Y, Inoue K, Kohara M
  • Journal Title: Biochem Biophys Res Commun. Volume: 23 Issue: 1 Pages: 56-62
  • DOI: 10.1016/j.bbrc.2014.04.053
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Suppression of hepatitis C virus replication by cyclin-dependent kinase inhibitors (2014)
  • Author(s): Tsubasa Munakata, Makoto Inada, Yuko Tokunaga, Takaji Wakita, Michinori Kohara, Akio Nomoto
  • Journal Title: Antiviral Research Volume: 108 Pages: 79-87
  • DOI: 10.1016/j.antiviral.2014.05.011
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Nonstructural Protein 5A Is Incorporated into Hepatitis C Virus Low-Density Particle through Interaction with Core Protein and Microtubules during Intracellular Transport (2014)
  • Author(s): Chao-Kuen Lai, Vikas Saxena, Chung-Hsin Tseng, King-Song Jeng, Michinori Kohara, Michael M. C. Lai
  • Journal Title: PLoS ONE Volume: 9 Issue: 6 Pages: e99022-e99022
  • DOI: 10.1371/journal.pone.0099022
  • Peer Reviewed / Open Access
• [Journal Article] Tupaia belangeri as an experimental animal model for viral infection (2014)
  • Author(s): Kyoko Tsukiyama-Kohara, Michinori Kohara
  • Journal Title: Experimental Animals Volume: 63 Pages: 367-374
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Targeted induction of interferon-λ in humanized chimeric mouse liver abrogates hepatotropic virus infection (2013)
  • Author(s): Nakagawa S, Hirata Y, Kameyama T, Tokunaga Y, Nishito Y, Hirabayashi K, Yano J, Ochiya T, Tateno C, Tanaka Y, Mizokami M, Tsukiyama-Kohara K, Inoue K, Yoshiba M, Takaoka A, Kohara M
  • Journal Title: PLoS One Volume: 8 Issue: 3 Pages: e59611-e59611
  • DOI: 10.1371/journal.pone.0059611
  • Peer Reviewed
• [Journal Article] A serine palmitoyltransferase inhibitor blocks hepatitis C virus replication in human hepatocytes. (2013)
  • Author(s): Katsume A, Tokunaga Y, Hirata Y, Munakata T, Saito M, Hayashi H, Okamoto K, Ohmori Y, Kusanagi I, Fujiwara S, Tsukuda T, Aoki Y, Klumpp K, Tsukivama-Kohara K, El-Gohary A, Sudoh M, Kohara M.
  • Journal Title: Gastroenterology Volume: 145(4) Issue: 4 Pages: 865-79
  • DOI: 10.1053/j.gastro.2013.06.012
  • Peer Reviewed
• [Journal Article] DNA vaccine expressing the non-structural proteins of hepatitis C virus diminishes the expression of HCV proteins in a mouse model (2013)
  • Author(s): Takeshi Wada, Michinori Kohara and Yasuhiro Yasutomi
  • Journal Title: Vaccine Volume: 31 Issue: 50 Pages: 5968-5974
  • DOI: 10.1016/j.vaccine.2013.10.037
  • Peer Reviewed
• [Journal Article] Isolation and characterization of highly replicable hepatitis C virus genotype 1a strain HCV-RMT (2013)
  • Author(s): Masaaki Arai, Yuko Tokunaga, Asako Takagia, Yoshimi Tobita, Yuichi Hirata, Yuji Ishida, Chise Tateno, Michinori Kohara
  • Journal Title: PLoSONE Volume: 8 Issue: 12 Pages: e82527-e82527
  • DOI: 10.1371/journal.pone.0082527
  • Peer Reviewed
• [Journal Article] Synthetic lipophilic antioxidant BO-653 suppresses HCV replication. (2012)
  • Author(s): Yasui F.
  • Journal Title: J. Medical Virology Volume: 85 Issue: 2 Pages: 241-249
  • DOI: 10.1002/jmv.23466
  • Peer Reviewed
• [Journal Article] Kinetics of peripheral hepatitis B virus-specific CD8(+) T cells in patients with onset of viral reactivation. (2012)
  • Author(s): Aoki J, Kowazaki Y, Ohtsuki T, Okamoto R, Ohashi K, Hayashi S, Sakamaki H, Kohara M, Kimura K
  • Journal Title: J Gastroenterol Volume: 47 Issue: 6 Pages: 22-22
  • DOI: 10.1007/s00535-012-0676-y
  • NAID: 10031184063
  • Peer Reviewed
• [Journal Article] Hepatitis C virus kinetics by administration of pegylated interferon-lin human and chimeric mice carrying human hepatocytes with variants of the IL28B gene (2012)
  • Author(s): Watanabe T, Sugauchi F, Tanaka Y, Matsuura K, Yatsuhashi H, Murakami S, Iijima S, Iio E, Sugiyama M, Shimada T, Kakuni M, Kohara M, Mizokami M
  • Journal Title: Gut Volume: Nov 7 Issue: 9 Pages: 1340-6
  • DOI: 10.1136/gutjnl-2012-302553
  • Peer Reviewed
• [Presentation] 選択的Wnt/β-catenin/CBPシグナル阻害剤による肝線維症改善作用 (2014)
  • Author(s): 徳永優子、木村公則、大槻貴博、林 幸子、原 詳子、宗方圭祐、比島恒和、小路弘行、小嶋聡一、小原道法
  • Organizer: 第62回日本ウイルス学会学術集会
  • Place of Presentation: パシフィコ横浜〔神奈川）
  • Year and Date: 2014-11-10 – 2014-11-12
• [Presentation] Inhibitory effects of pycnogenol on Hepatitis C Virus replication (2014)
  • Author(s): Sayeh Ezzikouri, Michinori Kohara, Kyoko Tsukiyama-Kohara
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Resistance to cyclosporine S derives from mutations in Hepatitis C virus nonstructural proteins (2014)
  • Author(s): Masaaki Arai, Kyoko Tsukiyama-Kohara, Asako Takagi, Yoshimi Tobita, Kazuaki Inoue, Michinori Kohara
  • Organizer: 21th International Symposium on Hepatitis C and Related Virus
  • Place of Presentation: Banff（Canada）
  • Year and Date: 2014-09-07 – 2014-09-11
• [Presentation] Selective inhibitor of Wnt/β-catenin/CBP signaling ameliorates hepatitis C virus-induced liver fibrosis (2014)
  • Author(s): Yuko Tokunaga, Kiminori Kimura, Takahiro Ohtsuki, Yukiko Hayashi, Mitsuko Hara, Keisuke Munekata, Tsunekazu Hishima, Soichi Kojima, Michinori Kohara
  • Organizer: 21th International Symposium on Hepatitis C and Related Virus
  • Place of Presentation: Banff（Canada）
  • Year and Date: 2014-09-07 – 2014-09-11
• [Presentation] Suppression of hepatitis C virus replication by cyclin-dependent kinase inhibitors (2014)
  • Author(s): Tubasa Munakata, Makoto Inada, Yuko Tokunaga, Takaji Wakita, Michinori Kohara, Akio Nomoto
  • Organizer: 21th International Symposium on Hepatitis C and Related Virus
  • Place of Presentation: Banff（Canada）
  • Year and Date: 2014-09-07 – 2014-09-11
• [Presentation] HBV Pathogenesis and host response in Tupaia belangeri (2014)
  • Author(s): Takahiro Sanada, Kyoko Tsukiyama-Kohara, S Ezzikouri, Naoki Yamamaoto, Michinori Kohara
  • Organizer: 2014 Internationl Meeting on Molecular Biology of Hepatitis B Viruses
  • Place of Presentation: Los Angeles (USA)
  • Year and Date: 2014-09-03 – 2014-09-06
• [Presentation] Development of Tupaia Belangeri for small animal infection model of hepatitis B virus, according to the genomic research (2014)
  • Author(s): Takahiro Sanada, Kyoko Tsukiyama-Kohara, S Ezzikouri, Naoki Yamamaoto, Michinori Kohara
  • Organizer: XVIth International Congress of Virology
  • Place of Presentation: Montreal (Canada)
  • Year and Date: 2014-07-27 – 2014-08-01
• [Presentation] Comprehensive analysis of B-lymphoma cells spontaneously developed in transgenic mice that express the full hepatitis C virus genome in B cells (2014)
  • Author(s): Kyoko Tsukiyama-Kohara, Michinori Kohara, Yuri Kasama
  • Organizer: XVIth International Congress of Virology
  • Place of Presentation: Montreal (Canada)
  • Year and Date: 2014-07-27 – 2014-08-01
• [Presentation] 免疫の賦活化によるウイルス性肝炎治療及び肝線維症治療に向けて (2014)
  • Author(s): 小原道法
  • Organizer: 第9回高輪ウイルス肝炎フォーラム
  • Place of Presentation: JCHO東京高輪病院 (東京)
  • Year and Date: 2014-06-14
  • Invited
• [Presentation] Non-DAAセリンパルミトイル基転移酵素阻害剤とDAA併用による薬剤耐性株の出現を押さえた強力なHCV複製阻害 (2014)
  • Author(s): 小原道法、徳永優子、須藤正幸
  • Organizer: 第50回日本肝臓学会総会
  • Place of Presentation: ホテルニューオータニ（東京）
  • Year and Date: 2014-05-29
• [Presentation] C型肝炎・肝硬変発症機序及びその制御を目指して (2014)
  • Author(s): 小原 道法
  • Organizer: 第22回桜山感染症研究会(
  • Place of Presentation: 名古屋
  • Invited
• [Presentation] ワクシニアLC16m8株（HCV,SARS,Influenza）ベクター (2013)
  • Author(s): 小原 道法
  • Organizer: 第17回日本ワクチン学会学集会
  • Place of Presentation: 三重
  • Invited
• [Remarks] 公益財団法人東京都医学総合研究所プロジェクト研究
  • URL: http://www.igakuken.or.jp/project/detail/infectious.html