Treatment of chronic hepatitis B for the prevention of development of hepatocellular carcinoma and analyses of hepatitis B virus insertional mutagenesis as a first hit during multistep liver carcinogenesis.

治疗慢性乙型肝炎以预防肝细胞癌的发展，并分析乙型肝炎病毒插入突变作为多步肝癌发生过程中的第一个打击。

• 批准号: 16590616
• 负责人: MINAMI Masahito
• 金额: $ 1.98万
• 依托单位: Graduate School of Medical Science, Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590616/
• 关键词: hepatitis B virus; chronic hepatitis; hepatocellular carcinoma; Alu-PCR; lamivudine; viral resistance; peptide nucleic acid

1.Lamivudine, a nucleoside analog, has been licensed for chronic hepatitis B in Japan since 2000. 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 developed a sensitive assay for these mutant viruses and analyzed their dynamics and mechanisms of emergence. We revealed that variant HBV with mutations at YMDD motif already exists as minority in some HBV carriers without previous lamivudine treatment and YMDD mutants appear as early as within 6 months after lamivudine administration. However, these mutants sometimes disappear spontaneously despite continuous lamivudine administration. We also performed sensitive quantitative assay for early detection of YMDD mutants and a threshold of 10^<2.7> copies/ml was suggested for predicting viral breakthrough during lamivudine treatment.2.Growing evidence demonstrates that HBV integration and resulting insertional mutagenesis plays an important role in cell growth or maintenance in hepatocellular carcinoma (HCC). We analyzed HBV integration in chronic hepatitis tissues without HCC to explore early genetic change due to HBV preceding the development of HCC by a few decades. We found that a few groups of liver cells with a same HBV integration expand in chronic hepatitis tissues and identified several genes affected by HBV insertion. These genes included one known tumor suppressor gene, three human homologs of drosophila genes that are critical for organ development, one putative oncogene and one recently found chemokine. These data provided firm evidence of HBV integration in hepatocytes at an early stage of chronic infection and revealed usefulness of the HBV-related virus tagging approach to identify genes associated with cell growth and maintenance.

1.拉米夫定是一种核苷类似物，自2000年以来已在日本获得治疗慢性乙型肝炎（chronic hepatitis B）的许可。据报道，长期使用该药物导致出现耐药病毒，病毒DNA聚合酶的酪氨酸-甲硫氨酸-天冬氨酸-天冬氨酸（YMDD）基序发生点突变，临床病程加重。我们开发了一种敏感的检测这些突变病毒，并分析了它们的动态和出现的机制。我们发现，变异的HBV与变异的YMDD基序已经存在的少数在一些HBV携带者没有以前的拉米夫定治疗和YMDD突变出现早在拉米夫定后6个月内。然而，这些突变体有时会自发消失，尽管连续拉米夫定管理。我们还进行了敏感的定量检测YMDD突变体的早期检测，并建议10 μ <2.7>copies/ml作为预测拉米夫定治疗期间病毒突破的阈值。2.越来越多的证据表明，HBV整合和由此产生的插入突变在肝细胞癌（HCC）细胞生长或维持中起重要作用。我们分析了HBV在慢性肝炎组织中的整合，以探讨HBV在肝癌发生前几十年的早期遗传变化。我们发现有几组具有相同HBV整合的肝细胞在慢性肝炎组织中扩增，并确定了受HBV插入影响的几个基因。这些基因包括一个已知的肿瘤抑制基因，三个果蝇基因的人类同源物，这些基因对器官发育至关重要，一个假定的致癌基因和一个最近发现的趋化因子。这些数据提供了在慢性感染的早期阶段HBV在肝细胞中整合的有力证据，并揭示了HBV相关病毒标记方法用于鉴定与细胞生长和维持相关的基因的有用性。

项目成果

Hepatitis B virus-related insertional mutagenesis in chronic hepatitis B patients as an early drastic genetic change leading to hepatocarcinogenesis

• DOI: 10.1038/sj.onc.1208628
• 发表时间: 2005-06-01
• 期刊: ONCOGENE
• 影响因子: 8
• 作者: Minami, M;Daimon, Y;Okanoue, T
• 通讯作者: Okanoue, T

Measurement of hepatitis B virus core-related antigen is valuable for identifying patients who are at low risk of lamivudine resistance

• DOI: 10.1111/j.1478-3231.2005.01200.x
• 发表时间: 2006-02-01
• 期刊: LIVER INTERNATIONAL
• 影响因子: 6.7
• 作者: Tanaka, E;Matsumoto, A;Kiyosawa, K
• 通讯作者: Kiyosawa, K

Hepatitis B virus reactivation in a patient undergoing steroid-free chemotherapy.

接受无类固醇化疗的患者体内乙型肝炎病毒重新激活。

• 发表时间: 2004
• 期刊: World Journal of Gastroenterology 10
• 作者: Shimizu D;Nomura K;Matsumoto Y;Ueda K;Yamaguchi K;Minami M;Itoh Y;Horiike S;Morita A;Taniwaki M;Okanoue T.
• 通讯作者: Okanoue T.

Update of research and management of hepatitis B

乙型肝炎研究和管理的最新进展

• 发表时间: 2006
• 期刊: Journal of Gastroenterology 41(2)
• 作者: Okanoue T;Minami M.
• 通讯作者: Minami M.

Prediction of breakthrough hepatitis due to lamivudine-resistant hepatitis B virus by a sensitive semiquantitative assay using peptide nucleic acids

通过使用肽核酸的灵敏半定量测定来预测拉米夫定耐药乙型肝炎病毒引起的突破性肝炎

• 发表时间: 2006
• 期刊: Intervirology 49(5)
• 作者: Mori K;Minami M;Kirishima T;Kunimoto K;Okita M;Nakayama M;Makiyama A;Yamaoka J;Nakajima T;Yasui K;Itoh Y;Okanoue T.
• 通讯作者: Okanoue T.