Analyzing the contribution of intracellular localization of hepatitis C virus replication complex for the drug resistance.

分析丙型肝炎病毒复制复合物的细胞内定位对耐药性的贡献。

• 批准号: 17590627
• 负责人: MAEKAWA Shinya
• 金额: $ 2.24万
• 依托单位: University of Yamanashi
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590627/
• 关键词: Hepatitis C Virus; NS5A protein; HCV replicon; interferon; インターフェロン耐性; C型肝炎ウイルス; レプリコン; 複製複合体; コレステロール; スフィンゴ脂質; 脂質ラフト

This research is aimed to disclose the relationship between intracellular localization of hepatitis C virus (HCV) replication complex and the effect of anti-viral drugs such as interferon-alpha, and finally to develop new anti-HCV strategies through understanding of viral replication mechanism(s) in the replication complex.Recently, micro-domains of organelle membranes rich in sphingomyelin and cholesterol in the form of "lipid rafts" have been considered to act as a scaffold for HCV replication complex formation, indicating the possibility that the lipid raft could be a new therapeutic target. In the present study, we investigated the effect of myriocin, an inhibitor of sphingomyelin synthesis, on HCV replication using an HCV-subgenomic replicon system, as well as the infectious HCV culture system. We also investigated the combined effect of myriocin plus interferon-alpha, or myriocin plus simvastatin, on HCV replication. Myriocin suppressed intracellular RNA replication of both genotype lb subgenomic HCV replicon RNA and genotype 2a infectious HCV RNA in a time-and dose-dependent manner (subgenomic HCV-lb, max. 70% at 120 hours ; genomic HCV-2a, max. 60% at 96 hours). Combination treatment with myriocin plus IFN or simvastatin attenuated intracellular HCV-RNA replication synergistically.Our data demonstrate that both the sphingomyelin synthesis inhibitor and HMG-CoA inhibitor strongly suppress HCV replication, irrespective of genotype, indicating that the lipid metabolism of the host could be a novel target for HCV therapy.

本研究旨在揭示丙型肝炎病毒（HCV）复制复合体的细胞内定位与抗病毒药物（如干扰素-α）的作用之间的关系，并最终通过了解复制复合体中病毒复制机制来开发新的抗HCV策略。富含鞘磷脂和胆固醇的细胞器膜的微结构域以“脂筏”的形式被认为是HCV复制复合物形成的支架，提示脂筏可能成为新的治疗靶点。在本研究中，我们调查的影响myriocin，鞘磷脂合成的抑制剂，HCV复制使用HCV亚基因组复制子系统，以及感染性HCV培养系统。我们还研究了多球菌素加α干扰素或多球菌素加辛伐他汀对HCV复制的联合作用。多粒壳菌素以时间和剂量依赖性方式抑制基因型Ib亚基因组HCV复制子RNA和基因型2a感染性HCV RNA的细胞内RNA复制（亚基因组HCV-Ib，max. 120小时时70%;基因组HCV-2a，最大值96小时时为60%）。多球壳菌素加IFN或辛伐他汀联合治疗协同减弱细胞内HCV-RNA复制，我们的数据表明，鞘磷脂合成抑制剂和HMG-CoA抑制剂强烈抑制HCV复制，无论基因型，表明宿主的脂质代谢可能是一个新的HCV治疗靶点。

项目成果

Negative regulation of intracellular hepatitis C virus replication by interferon regulatory factor 3

• DOI: 10.1007/s00535-006-1842-x
• 发表时间: 2006-08
• 期刊: Journal of Gastroenterology
• 影响因子: 6.3
• 作者: T. Yamashiro;N. Sakamoto;M. Kurosaki;N. Kanazawa;Y. Tanabe;M. Nakagawa;Cheng-Hsin Chen;Yasuhiro Itsui-Ya

Divergent activities of interferon-alpha subtypes against intracellular hepatitis C virus replication

• DOI: 10.1016/j.hepres.2005.10.005
• 发表时间: 2006-01-01
• 期刊: HEPATOLOGY RESEARCH
• 影响因子: 4.2
• 作者: Koyama, T;Sakamoto, N;Watanabe, M
• 通讯作者: Watanabe, M

Site-specific nutation of the interferon sensitivity-determining region (ISDR) modulates hepatitis C virus replication.

干扰素敏感性决定区 (ISDR) 的位点特异性章动调节丙型肝炎病毒复制。

• 发表时间: 2006
• 期刊: Journal of Viral Hepatitis Sep;13(9)
• 作者: Kohashi T;Maekawa S;Enomoto N;et al.
• 通讯作者: et al.

Site-specific mutation of the interferon sensitivity-determining region (ISDR) modulates hepatitis C virus replication

• DOI: 10.1111/j.1365-2893.2006.00739.x
• 发表时间: 2006-09-01
• 期刊: JOURNAL OF VIRAL HEPATITIS
• 影响因子: 2.5
• 作者: Kohashi, T.;Maekawa, S.;Watanabe, M.
• 通讯作者: Watanabe, M.

Expressional screening of interferon-stimulated genes for antiviral activity against hepatitis C virus replication

• DOI: 10.1111/j.1365-2893.2006.00732.x
• 发表时间: 2006-10-01
• 期刊: JOURNAL OF VIRAL HEPATITIS
• 影响因子: 2.5
• 作者: Itsui, Y.;Sakamoto, N.;Watanabe, M.
• 通讯作者: Watanabe, M.