Development of a novel three-dimensional culture system for hepatitis viruses

• Project/Area Number: 22790664
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Gastroenterology
• Research Institution: Nagoya City University
• Principal Investigator: MURAKAMI Shuko 名古屋市立大学, 大学院・医学研究科, 助教 (50454848)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2012: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: HBV / 肝細胞 / 3 次元培養 / キメラマウス / 肝炎 / ウイルス / 感染症 / 3次元培養 / 細胞・組織

Research Abstract

Investigation of hepatitis B virus (HBV) has been hampered by the absence of accessible in vitro modeling tools. The aim of this study is to design an in vitro model of HBV infection that would support long-term replication. Immortalized normal human hepatocytes, or primary human hepatocytes isolated from chimeric mice with human liver were cultured with three-dimensional (3D) culture system to obtain a tissue-like formation and exposed to HBV-infected serum samples. The HBV-DNA level in supernatants of the culture was higher than 10^5 copies/ml and continued to be detectable for >1 month. The extracellular HBsAg was detected during the culture period. The 3D culture system with hepatocytes provides an accessible in vitro tool to investigate HBV cell entry, replication and susceptibility to new anti-viral agents for multiple HBV genotypes/subgenotypes.

Research Products

• [Journal Article] Mechanism of the Dependence of Hepatitis B Virus Genotype G on Co-infection with Other Genotypes for Viral Replication (2013)
  • Author(s): Sakamoto T, Tanaka Y, Kani S, Sugiyama M, Watanabe T, Iijima S, Murakami S, Matsuura K, Kusakabe A, Shinkai N, Sugauchi F, Mizokami M
  • Journal Title: J Viral. Hepat Volume: 20 (4) Issue: 4 Pages: 27-36
  • DOI: 10.1111/jvh.12022
  • 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
• [Journal Article] Multiple intra-familial transmission patterns of hepatitis B virus genotype D in north-eastern Egypt (2012)
  • Author(s): Ragheb M, Elkady A, Tanaka Y
  • Journal Title: J Med Virol. Volume: 84(4) Issue: 4 Pages: 587-595
  • DOI: 10.1002/jmv.23234
  • Peer Reviewed
• [Journal Article] Cross-species transmission of gibbon and orangutan hepatitis B virus to uPA/SCID mice with human hepatocytes (2011)
  • Author(s): Sa-nguanmoo P, Tanaka Y, Ratanakorn P, Sugiyama M, Murakami S, Payungporn S, Sommanustweechai A, Mizokami M, Poovorawan Y
  • Journal Title: Virus Res Volume: 158 (1-2) Issue: 1-2 Pages: 209-15
  • DOI: 10.1016/j.virusres.2011.04.002
  • Peer Reviewed
• [Journal Article] Cross-species transmission of gibbon and orangutan hepatitis Bvirus to uPA/SCID mice with human hepatocytes. (2011)
  • Author(s): Sa-Nguanmoo P, Murakami S.
  • Journal Title: Virus Research Volume: (印刷中)
  • Peer Reviewed
• [Presentation] IL28B genetic variants and serum IP-10 level associated with virological response to PEG-IFN/RBV and PEG-IFN/RBV/Telaprevir therapy (2012)
  • Author(s): Matsuura K, Tanaka Y, Watanabe T, Murakami S, Iio E, Endo M, Shinkai N, Fujiwara K, Miyaki T, Nojiri S, Kusakabe A, Orito E, Joh T, Mizokami M
  • Organizer: The 63rd Annual Meeting of the American Association for the Study of Liver Diseases
  • Place of Presentation: Boston in USA
• [Presentation] 核酸アナログを投与した B 型慢性肝炎患者における interferoninducible protein-10値の動態 (2012)
  • Author(s): 新海登, 松浦健太郎,渡邊綱正,村上周子,宮木知克,藤原圭,日下部篤宣,飯尾悦子,野尻俊輔,城卓志,田中靖人
  • Organizer: 第20回日本消化器関連学会週間
  • Place of Presentation: 神戸市
• [Presentation] 末梢血単核球を用いた C 型慢性肝炎患者PEG-IFN/RBV 投与直後の ISG 挙動 (2012)
  • Author(s): 飯島沙幸, 渡邊 綱正,松浦健太郎,飯尾悦子,新海登,村上周子,田中靖人
  • Organizer: 第20回日本消化器関連学会週間
  • Place of Presentation: 神戸市
• [Presentation] 2 剤,3 剤併用療法における,IP-10の治療前,治療開始早期の推移と治療効果,ウイルス動態に与える影響 (2012)
  • Author(s): 松浦健太郎, 飯尾悦子,日下部篤宣,新海 登,藤原 圭,宮木知克,野尻俊輔,村上周子,渡邊綱正,折戸悦朗,城卓志,田中靖人
  • Organizer: 第20回日本消化器関連学会週間
  • Place of Presentation: 神戸市