Analysis of hepatitis virus genomes using human hepatocytes transplanted chimeric mice and ultra-deep sequencing.

• Project/Area Number: 24790701
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Hiroshima University
• Principal Investigator: ABE Hiromi 広島大学, 医歯薬保健学研究院(医), 准教授 (70572329)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: ヒト肝細胞キメラマウス / 肝炎ウイルス / 次世代シーケンサー

Research Abstract

Direct-acting antiviral agents (DAAs) against hepatitis C virus (HCV) have been developed. The aim of this study is to investigate efficacy of combinations of NS3 inhibitor with NS5A or NS5B inhibitor and track the emergence of resistant strains. Human hepatocyte chimeric mice were infected with serum from HCV patients or infectious clones carrying resistance mutations and treated with combinations of DAAs. Nucleotide and amino acid sequences were analyzed by both direct and ultra-deep sequencing. Mice infected with a clone with each DAAs resistant mutations responded poorly to NS5A inhibitor and NS3 inhibitor combination therapy. Although HCV RNA became undetectable soon after the beginning of combination therapy with NS5A and NS5B inhibitor, rebound with emergence of resistance against all three drugs occurred. Our results showed that resistant strains easily develop from cloned virus strains by mutation.

Research Products

• [Journal Article] Ultradeep sequencing study of chronic hepatitis C virus genotype 1 infection in patients treated with daclatasvir, peginterferon, and ribavirin (2014)
  • Author(s): Murakami, E. Imamura, M. Hayes, C. N. Abe, H. Hiraga, N. Honda, Y. Ono, A. Kosaka, K. Kawaoka, T. Tsuge, M. Aikata, H. Takahashi, S. Miki, D. Ochi, H. Matsui, H. Kanai, A. Inaba, T. McPhee, F. Chayama, K.
  • Journal Title: Antimicrob Agents Chemother Volume: 58 Issue: 4 Pages: 2105-2112
  • DOI: 10.1128/aac.02068-13
  • Peer Reviewed / Open Access
• [Journal Article] HLA-DQB1*03 Confers Susceptibility to Chronic Hepatitis C in Japanese: A Genome-Wide Association Study. (2013)
  • Author(s): Miki D, Ochi H, Takahashi A, Hayes CN, Urabe Y, Abe H, Kawaoka T, Tsuge M, Hiraga N, Imamura M, Kawakami Y, Aikata H, Takahashi S, Akuta N, Suzuki F, Ikeda K, Kumada H, Karino Y, Toyota J, Tsunoda T, Kubo M, Kamatani N, Nakamura Y, Chayama K.
  • Journal Title: PLoS One. Volume: 8 Issue: 12 Pages: e84226-e84226
  • DOI: 10.1371/journal.pone.0084226
  • Peer Reviewed
• [Journal Article] Changes in mRNA expression of hypothalamic appetite-regulating peptides during different phases of lactation in rats (2013)
  • Author(s): Suzuki Y,Nakahara K, Maruyama K, Okame R, Ensho T, Inoue Y,Murakami N.
  • Journal Title: J Mol Endocrinol Volume: 52 Issue: 1 Pages: 97-109
  • DOI: 10.1016/j.bbrc.2013.10.040
  • Peer Reviewed
• [Journal Article] A translational study of resistance emergence using sequential direct-acting antiviral agents for hepatitis C using ultra-deep sequencing (2013)
  • Author(s): Abe H, Hayes CN, Hiraga N, Imamura M, Tsuge M, Miki D, Takahashi S, Ochi H, Chayama K
  • Journal Title: Am J Gastroenterol Volume: 108 Issue: 9 Pages: 1464-1472
  • DOI: 10.1038/ajg.2013.205
  • Peer Reviewed
• [Journal Article] A translational study of resistance emergence using sequential direct-acting antiviral agents for hepatitis C using ultra-deep sequencing. (2013)
  • Author(s): Abe H, Hayes CN, Hiraga N, Imamura M, Tsuge M, Miki D, Takahashi S, Ochi H, Chayama K.
  • Journal Title: Am J Gastroenterol. Volume: 108 Issue: 7 Pages: 1464-1472
  • DOI: 10.1038/ajg.2013.20
  • Peer Reviewed
• [Presentation] Expression levels of IFNL4 and other IFN.s are determined by dinucleotide polymorphism of IFNL4 and correlate with basal expression levels of interferon stimulated genes and effect of interferon (2013)
  • Author(s): Abe H. Hayes CN, Hiraga N, Imamura M, Miki D, Tsuge M, Ochi H, Chayama K
  • Organizer: The 64th AASLD Annual Meeting
  • Place of Presentation: Washington DC, USA(Oral)
• [Presentation] ヒト肝細胞キメラマウスと次世代シーケンサーを用いた新規抗ウイルス薬に対するHCVゲノムの解析 (2013)
  • Author(s): 阿部弘美 , C. Nelson Hayes,平賀伸彦 ,今村道雄 ,柘植雅貴 ,三木大樹 3,山田一麿呂,高橋祥一 ,越智秀典 ,茶山一彰
  • Organizer: 第49回日本肝臓学会総会
  • Place of Presentation: 東京(口頭発表)
• [Presentation] ヒト肝細胞キメラマウスと次世代シーケンサーを用いた新規抗ウイルス薬に対するHCVゲノムの解析 (2013)
  • Author(s): 阿部弘美, C. Nelson Hayes, 平賀伸彦, 今村道雄, 柘植雅貴, 三木大樹, 山田一麿呂, 高橋祥一, 越智秀典, 茶山一彰
  • Organizer: 日本肝臓学会総会
  • Place of Presentation: 東京
• [Presentation] Expression levels of IFNL4 and other IFN-λs are determined by dinucleotide polymorphism of IFNL4 and correlate with basal expression levels of interferon stimulated genes and effect of interferon (2013)
  • Author(s): Abe H. Hayes CN, Hiraga N, Imamura M, Miki D, Tsuge M, Ochi H, Chayama K
  • Organizer: The 64th AASLD Annual Meeting
  • Place of Presentation: アメリカ（ボストン）
• [Presentation] Analysis of HCV genomes against direct-acting antiviral agents using human hepatocyte chimeric mice and ultra-deep sequencing (2012)
  • Author(s): 阿部弘美、平賀伸彦、今村道雄、Hayes CN、三木大樹、柘植雅貴、高橋祥一、越智秀典、茶山一彰
  • Organizer: AASLD Annual Meeting 2012
  • Place of Presentation: Boston, USA
• [Remarks]
  • URL: http://home.hiroshima-u.ac.jp/naika1/
• [Remarks] 広島大学大学院医歯薬保健学研究院 応用生命科学部門 消化器・代謝内科学
  • URL: http://home.hiroshima-u.ac.jp/naika1/