Treatment of hepatitis B virus without inducing antiviral resistant viruses and analyses of HBV integration at an early stage of infection before development of hepatocellular carcinoma.

• Project/Area Number: 14570495
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Graduate School of Medical Science, Kyoto Prefectural University of Medicine
• Principal Investigator: MINAMI Masahito Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Research Associate, 大学院・医学研究科, 助手 (60326220)
• Co-Investigator(Kenkyū-buntansha): OKANOUE Takeshi Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Professor, 大学院・医学研究科, 教授 (20150568)
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥3,400,000 (Direct Cost: ¥3,400,000); Fiscal Year 2003: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2002: ¥2,300,000 (Direct Cost: ¥2,300,000)
• Keywords: hepatitis B virus / chronic hepatitis / hepatocellular carcinoma / Alu-PCR / lamivudine / viral resistance / peptide nucleic acid / ウイルス組み込み / 肝発癌 / lamivudine / 薬剤耐性ウイルス / PCR clamping / 薬剤耐性 / HBV

Research Abstract

1.Lamivudine, a nucleoside analog, is recently used for treatment of HBV infection. However, it is reported that prolonged administration of this drug results in emergence of resistant viruses with point mutations at tyrosine-methionine-aspartate-aspartate (YMDD) motif of viral DNA polymerase and exacerbation of clinical course. We have developed a sensitive assay for these mutant viruses and analyzed emergence of variant viruses before and during lamivudine treatment to clarify mechanisms involved in outbreak of YMDD-mutant viruses. We have demonstrated that variant HBV with mutations at YMDD motif already exists as minority in some HBV carriers without lamivudine treatment. However, existence of these mutant viruses did not always lead to their outbreak during lamivudine treatment. These results imply that outbreak of YMDD mutants needs other selection forces or factors, possibly, mutations in other regions than YMDD motif, impaired host immunity to HBV, and such.
2.Growing evidence demonstrates that hepatitis B virus (HBV) integration and resulting insertional mutagenesis play an important role in cell growth or maintenance in hepatocellular carcinomas (HCCs). We previously reported that HBV integration occurs early during HBV infection, even after acute self-limiting hepatitis. Insertional mutagenesis may, therefore, represent the first drastic genetic change preceding the development of HCC by a few decades. Our virus-tagging approach provided (a) firm evidence of HBV integration in hepatocytes at an early stage of chronic infection and (b) revealed cellular genes possibly affected by HBV integration and related to liver tumorigenesis. In contrast to the previous consensus, we demonstrated a preferred chromosome for HBV integration which implies a selection mechanism for some integrants.

Research Products

• [Journal Article] Hepatitis B virus DNA in liver, serum, and peripheral blood mononuclear cells after the clearance of serum hepatitis B virus surface antigen. (2004)
  • Author(s): Murakami Y, Minami M, Daimon Y, Okanoue T.
  • Journal Title: Journal of Medical Virology 72 Pages: 203-214
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Hepatitis B virus reactivation in a patient undergoing steroid-free chemotherapy. (2004)
  • Author(s): Shimizu D, Nomura K, Matsumoto Y, Ueda K, Yamaguchi K, Minami M, Itoh Y, Horiike S, Morita A, Taniwaki M, Okanoue T.
  • Journal Title: World Journal of Gastroenterology 10 Pages: 2301-2302
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Hepatitis B virus DNA in liver, serum, and peripheral blood mononuclear cells after the clearance of serum hepatitis B virus surface antigen. (2004)
  • Author(s): Murakami Y, Minami M, Daimon Y, Okanoue T
  • Journal Title: Journal of Medical Virology 72 Pages: 203-214
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Hepatitis B virus reactivation in a patient undergoing steroid-free chemotherapy. (2004)
  • Author(s): Shimizu D, Nomura K, Matsumoto Y, Ueda K, Yamaguchi K, Minami M, Itoh Y, Horiike S, Morita A, Taniwaki M, Okanoue T
  • Journal Title: World Journal of Gastroenterology 10 Pages: 2301-2302
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Lamivudine therapy for Japanese patients with cirrhosis B. (2003)
  • Author(s): Okanoue T, Mori K, Kirishima T, Kunimoto K, Yasui K, Itoh Y, Minami M.
  • Journal Title: Intervirology 46 Pages: 394-399
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Detection of YMDD mutant using a novel sensitive method in chronic liver disease type B patients before and during lamivudine treatment. (2002)
  • Author(s): Kirishima T, Okanoue T, Daimon Y, Itoh Y, Nakamura H, Morita A, Toyama T, Minami M.
  • Journal Title: Journal of Hepatology 37 Pages: 259-265
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Hepatitis B virus-related insertional mutagenesis in chronic hepatitis B patients as an early drastic genetic change leading to hepatocarcinogenesis.
  • Author(s): Minami M, Daimon Y, Mori K, Takashima H, Nakajima T, Itoh Y, Okanoue T.
  • Journal Title: Oncogene (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Hepatitis B virus-related insertional mutagenesis in chronic hepatitis B patients as an early drastic genetic change leading to hepatocarcinogenesis.
  • Author(s): Minami M, Daimon Y, Mori K, Takashima H, Nakajima T, Itoh Y, Okanoue T
  • Journal Title: Oncogene (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Patent(Industrial Property Rights)] 特許権 (2002)
  • Inventor(s): 南 祐仁, 桐島寿彦, 岡上 武, 向出雅一
  • Industrial Property Rights Holder: 株式会社エスアールエル
  • Industrial Property Number: 2002-244392
  • Filing Date: 2002-08-23
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Murakami Y et al.: "Hepatitis B virus DNA in liver, serum, and peripheral blood mononuclear cells after the clearance of serum hepatitis B virus surface antigen."Journal of Medical Virology. 72. 203-214 (2004)
• [Publications] Okanoue T et al.: "Lamivudine therapy for Japanese patients with cirrhosis B."Intervirology. 46・6. 394-399 (2003)
• [Publications] Kirishima et al.: "Detection of YMDD mutant using a novel sensitive method in chronic liver disease type B patients before and during lamivudine treatment."Journal of Hepatology. 37・2. 259-265 (2002)
• [Publications] Okanoue T, Itoh Y, Kirishima T, Daimon Y, Toyama T, Morita A, Nakajima T, Minami M.: "Transient biochemical response in interferon therapy decreases the development of hepatocellular carcinoma for five years and improves the long-term survival of chronic hepatitis C patients"Hepatol Res. 23・1. 62-77 (2002)
• [Publications] Kirishima T, Okanoue T, Daimon Y, Itoh Y, Nakamura H, Morita A, Toyama T, Minami M.: "Detection of YMDD mutant using a novel sensitive method in chronic liver disease type B patients before and during lamivudine treatment"J Hepatol. 37・2. 259-265 (2002)