肝炎ウイルス複製を支持する新規肝癌由来培養細胞の樹立

2015 Fiscal Year Annual Research Report

• Project/Area Number: 26670378
• Research Institution: Kanazawa University
• Principal Investigator: 本多 政夫 金沢大学, 保健学系, 教授 (00272980)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: ウイルス / 癌 / 感染症

Outline of Annual Research Achievements

現在、我が国のHCV 感染の多くを占めるゲノタイプ1b型HCVの細胞培養系は存在せず、出現する薬剤耐性ウイルスの解析や薬剤耐性ウイルスに対する効果的なDAAの選択には、ゲノタイプ1b型HCVの培養細胞系の確立が必要である。本研究目的は、困難とされているゲノタイプ1b型HCVの培養系を確立し、DAA による効果的なHCV 治療を確立する上で必修の基盤的研究を行うことを目的とした。ゲノタイプ１ｂ型HCV感染症例の肝細胞癌手術検体から癌幹細胞マーカーEpCAM陽性細胞分画を単離し、培養皿で継代が可能な細胞株を樹立した（KH細胞）。KH細胞は当初HCV-RNA陽性を示したが、継体を続けるごとにHCV-RNAは減少し、検出されなくなった。しかしながら、JFH-1株やHJ3-5株（構造領域がH77由来、非構造領域がJFH株由来）のRNAを導入するとHCVが効率よく複製することが確認された。ウイルス蛋白の発現も免疫染色やウエスタンブロットで検出された。HCVccによる感染実験でもKH細胞のHCV感染を確認した。網羅的遺伝子発現やカリオタイプの解析からKH細胞はHuh-7細胞、Huh-7.5とは完全に異なる細胞であることが明らかとなった。KH細胞はRIG-Iの変異を有さず、p53も野生型であった。またNOD-SCIDマウスに効率よく腫瘍を形成した。以上より、HCV感染・複製を支持する新規肝癌培養細胞株KH細胞の樹立に成功した。KH細胞にゲノタイプ1b型HCV感染者3例から得られた血清を用いて様々な条件下で感染実験を行った。ビタミンEはHCV感染・複製効率を飛躍的に亢進させることが報告されているが、ビタミンE存在下に於いてもKH細胞でのHCV複製は確認されなかった。

Research Products

• [Journal Article] Post-progression survival and progression-free survival in patients with advanced hepatocellular carcinoma treated by sorafenib. (2016)
  • Author(s): Terashima T, Yamashita T, Takata N, Nakagawa H, Toyama T, Arai K, Kitamura K, Yamashita T, Sakai Y, Mizukoshi E, Honda M, Kaneko S.
  • Journal Title: Hepatol Res. Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1111/hepr.12601.
  • Peer Reviewed / Open Access
• [Journal Article] Small tRNA-derived RNAs are increased and more abundant than microRNAs in chronic hepatitis B and C. (2015)
  • Author(s): Selitsky SR, Baran-Gale J, Honda M, Yamane D, Masaki T, Fannin EE, Guerra B, Shirasaki T, Shimakami T, Kaneko S, Lanford RE, Lemon SM, Sethupathy P.
  • Journal Title: Sci Rep. Volume: 5 Pages: 7675
  • DOI: 10.1038/srep07675.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Characteristics of hepatic fatty acid compositions in patients with nonalcoholic steatohepatitis. (2015)
  • Author(s): Yamada K, Mizukoshi E, Sunagozaka H, Arai K, Yamashita T, Takeshita Y, Misu H, Takamura T, Kitamura S, Zen Y, Nakanuma Y, Honda M, Kaneko S.
  • Journal Title: Liver Int. Volume: 35 Pages: 582-590
  • DOI: 10.1111/liv.12685.
  • Peer Reviewed / Open Access
• [Journal Article] Genome-wide association study identifies a PSMD3 variant associated with neutropenia in interferon-based therapy for chronic hepatitis C. (2015)
  • Author(s): Iio E, Matsuura K, Nishida N, Maekawa S, Enomoto N, Nakagawa M, Sakamoto N, Yatsuhashi H, Kurosaki M, Izumi N, Hiasa Y, Masaki N, Ide T, Hino K, Tamori A, Honda M, Kaneko S, Tanaka Y. et al
  • Journal Title: Hum Genet. Volume: 134 Pages: 279-89
  • DOI: 10.1007/s00439-014-1520-7.
  • Peer Reviewed / Open Access
• [Journal Article] TSU-68 Ameliorates Hepatocellular Carcinoma Growth by Inhibiting Microenvironmental Platelet-derived Growth Factor Signaling. (2015)
  • Author(s): Hara Y, Yamashita T, Oishi N, Nio K, Hayashi T, Nomura Y, Yoshida M, Hayashi T, Hashiba T, Asahina Y, Kondo M, Okada H, Sunagozaka H, Honda M, Kaneko S.
  • Journal Title: Anticancer Res. Volume: 35 Pages: 1423-1431
  • Peer Reviewed / Open Access
• [Journal Article] Inflammatory features of pancreatic cancer highlighted by monocytes/macrophages and CD4+ T cells with clinical impact. (2015)
  • Author(s): Komura T, Sakai Y, Harada K, Kawaguchi K, Takabatake H, Kitagawa H, Wada T, Honda M, Ohta T, Nakanuma Y, Kaneko S.
  • Journal Title: Cancer Sci. Volume: 106 Pages: 672-686
  • DOI: 10.1111/cas.12663.
  • Peer Reviewed / Open Access
• [Journal Article] Inhibition of microRNA-214 ameliorates hepatic fibrosis and tumor incidence in platelet-derived growth factor C transgenic mice. (2015)
  • Author(s): Okada H, Honda M, Campbell JS, Takegoshi K, Sakai Y, Yamashita T, Shirasaki T, Takabatake R, Nakamura M, Tanaka T, Kaneko S.
  • Journal Title: Cancer Sci. Volume: 106 Pages: 1143-1152
  • DOI: 10.1111/cas.12730.
  • Peer Reviewed / Open Access
• [Journal Article] Response to Importance of confounding factors in assessing fatty acid compositions in patients with non-alcoholic steatohepatitis. (2015)
  • Author(s): Yamada K, Mizukoshi E, Sunagozaka H, Arai K, Yamashita T, Takeshita Y, Misu H, Takamura T, Kitamura S, Zen Y, Nakanuma Y, Honda M, Kaneko S.
  • Journal Title: Liver Int. Volume: 35 Pages: 1773
  • DOI: 10.1111/liv.12755.
  • Peer Reviewed / Open Access
• [Journal Article] Autoantibody status and histological variables influence biochemical response to treatment and long-term outcomes in Japanese patients with primary biliary cirrhosis. (2015)
  • Author(s): Nakamura M, Kondo H, Tanaka A, Komori A, Ito M, Yamamoto K, Ohira H, Zeniya M, Hashimoto E, Honda M, Kaneko S, Ueno Y, Kikuchi K, Shimoda S, Harada K, Arai K, Miyake Y, Abe M, Taniai M, Saibara T, Sakisaka S, Takikawa H, Onji M, Tsubouchi H, Nakanuma Y, Ishibashi H.
  • Journal Title: Hepatol Res. Volume: 45 Pages: 846-855
  • DOI: 10.1111/hepr.12423.
  • Peer Reviewed / Open Access
• [Journal Article] Blood neutrophil to lymphocyte ratio as a predictor in patients with advanced hepatocellular carcinoma treated with hepatic arterial infusion chemotherapy. (2015)
  • Author(s): Terashima T, Yamashita T, Iida N, Yamashita T, Nakagawa H, Arai K, Kitamura K, Kagaya T, Sakai Y, Mizukoshi E, Honda M, Kaneko S.
  • Journal Title: Hepatol Res. Volume: 45 Pages: 949-959
  • DOI: 10.1111/hepr.12436.
  • Peer Reviewed / Open Access
• [Journal Article] Defeating EpCAM(+) liver cancer stem cells by targeting chromatin remodeling enzyme CHD4 in human hepatocellular carcinoma. (2015)
  • Author(s): Nio K, Yamashita T, Okada H, Kondo M, Hayashi T, Hara Y, Nomura Y, Zeng SS, Yoshida M, Hayashi T, Sunagozaka H, Oishi N, Honda M, Kaneko S.
  • Journal Title: J Hepatol. Volume: 63 Pages: 1164-1172
  • DOI: 10.1016/j.jhep.2015.06.009.
  • Peer Reviewed / Open Access
• [Journal Article] Transcriptomic Analysis of Chronic Hepatitis B and C and Liver Cancer Reveals MicroRNA-Mediated Control of Cholesterol Synthesis Programs. (2015)
  • Author(s): Selitsky SR, Dinh TA, Toth CL, Kurtz CL, Honda M, Struck BR, Kaneko S, Vickers KC, Lemon SM, Sethupathy P.
  • Journal Title: MBio. Volume: 6 Pages: e01500-e01515
  • DOI: 10.1128/mBio.01500-15.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Hepatitis B virus X protein stimulates HBV replication by regulating transcription factors associated with DNA or histone methylation (2015)
  • Author(s): Naoki Oishi, Xuyang Wang, Kazunori Kawaguchi, Masao Honda, Seishi Murakami, Shuichi Kaneko
  • Organizer: AASLD The Liver Meeting 2015
  • Place of Presentation: サンフランシスコ（アメリカ）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research
• [Presentation] Peretinoin, an acyclic retinoid, suppresses steatohepatitis and development of tumorigenesis by activated Atg16L1-dependent autophagy in mice fed an atherogenic high-fat diet (2015)
  • Author(s): Hikari Okada, Masao Honda, Kai Takegoshi, Taro Yamashita, Naoto Matsuzawa,Toshinari Takamura, Takuji Tanaka, Shuichi Kaneko
  • Organizer: AASLD The Liver Meeting 2015
  • Place of Presentation: サンフランシスコ（アメリカ）
  • Year and Date: 2015-11-13 – 2015-11-17
  • Int'l Joint Research
• [Presentation] Effective prevention of direct-acting antiviral-resistant hepatitis C virus by combination with LNA-anti-miR-122 therapy in cell culture (2015)
  • Author(s): Fanwei Liu, Tetsuro Shimakami, Kazuhisa Murai, Takayuki Shirasaki, Masaya Funaki, Masao Honda, Hong Tang, Seishi Murakami, Shuichi Kaneko
  • Organizer: 22nd International Symposium on Hepatitis C Virus and Related Viruses
  • Place of Presentation: ストラスブール（フランス）
  • Year and Date: 2015-10-09 – 2015-10-13
  • Int'l Joint Research