Suppression of hepatitis C virus replication by cyclosporine A

环孢菌素 A 抑制丙型肝炎病毒复制

• 批准号: 16590580
• 负责人: OOOKA Shinya
• 金额: $ 2.3万
• 依托单位: Tokyo Medical and Dental University
• 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-16590580/
• 关键词: Cyclosporin A; HCV replicon system; cyclophilin; 抗ウイルス療法; レトロウイルスベクター; サイクロフィリン; small interfering RNA

Cyclosporin A specifically suppresses hepatitis C virus replication in vitro at clinically achievable concentrations. In this study, we investigated the mechanisms of action of cyclosporin A against HCV replication. The in-vitro effects of cyclosporin A on HCV replication were analyzed using HCV replicon system that expresses chimeric luciferase reporter protein. The significant effects of cyclosporin A on expression of an HCV replicon, and the absence of such effects of FK506 which shares mechanisms of action with cyclosporin A, suggested the involvement of intracellular ligands of cyclosporin A, the cyclophilins. Transient and stable knock-down of the expression of cytoplasmic cyclophilins A, B and C by shRNA-expressing vectors suppressed HCV replication significantly. A cyclosporin analogue, cyclosporin D, which lacks immunosuppressive activity but exhibits cyclophilin binding, induced a similar suppression of HCV replication. Furthermore, cyclosporin A treatment of Huh7 cells induced an unfolded protein response exemplified by expression of cellular BiP/GRP78. Treatment of cells with thapsigargin and mercaptoethanol, which induce the unfolded protein responses, suppressed HCV replication, suggesting that the cyclosporin-induced unfolded protein responses mignt contribute to the suppression of HCV protein processing and replication. The anti-HCV activity of cyclosporin A is mediated through a specific blockade of cyclophilins and these molecules may constitute novel targets for anti-HCV therapeutics.

环孢素A在临床可达到的浓度下特异性抑制丙型肝炎病毒的体外复制。在这项研究中，我们研究了环孢素A对HCV复制的作用机制。利用表达嵌合荧光素酶报告蛋白的HCV复制子系统分析环孢素A对HCV体外复制的影响。环孢素A对HCV复制子的表达有显著影响，而FK506的作用机制与环孢素A相同，但FK506没有这种影响，这表明环孢素A参与了细胞内配体，即亲环蛋白。shrna表达载体瞬时稳定地敲除细胞质亲环蛋白A、B和C的表达可显著抑制HCV复制。环孢素类似物环孢素D缺乏免疫抑制活性，但表现出亲环蛋白结合，诱导了类似的HCV复制抑制。此外，环孢素A处理Huh7细胞诱导了未折叠蛋白反应，例如细胞BiP/GRP78的表达。用thapsigarin和巯基乙醇处理细胞，可诱导未折叠蛋白反应，抑制HCV复制，提示环孢素诱导的未折叠蛋白反应可能有助于抑制HCV蛋白的加工和复制。环孢素A的抗hcv活性是通过特异性阻断亲环蛋白介导的，这些分子可能成为抗hcv治疗的新靶点。

项目成果

Introduction of NS5A mutations enables subgenomic HCV replicon derived from chimpanzee‐infectious HC‐J4 isolate to replicate efficiently in Huh‐7 cells

• DOI: 10.1111/j.1365-2893.2004.00525.x
• 发表时间: 2004-09
• 期刊: Journal of Viral Hepatitis
• 影响因子: 2.5
• 作者: S. Maekawa;N. Enomoto;N. Sakamoto;M. Kurosaki;E. Ueda;T. Kohashi;H. Watanabe;C.‐H. Chen;T. Yamashiro;Y. Tanabe;N. Kanazawa;M. Nakagawa;C. Sato;M. Watanabe

Specific inhibition of hepatitis C virus replication by cyclosporin A

• DOI: 10.1016/j.bbrc.2003.11.080
• 发表时间: 2004-01-02
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Nakagawa, M;Sakamoto, N;Watanabe, M
• 通讯作者: Watanabe, M

Synergistic inhibition of intracellular hepatitis C virus replication by combination of ribavirin and interferon-alpha

利巴韦林和干扰素-α组合协同抑制细胞内丙型肝炎病毒复制

• 发表时间: 2004
• 期刊: J Infect Dis 189
• 作者: Sakamoto N;et al.
• 通讯作者: et al.

Down-regulation of p27^<Kip1> promotes cell proliferation of rat neonatal cardiomyocytes induced by nuclear expression of cyclinD1 and CDK4 : Evidence for impaired Skp2-dependent degradation of p27 in terminal differentiation.

p27^<Kip1> 的下调促进由细胞周期蛋白 D1 和 CDK4 的核表达诱导的大鼠新生心肌细胞的细胞增殖：终末分化中 p27 的 Skp2 依赖性降解受损的证据。

• 发表时间: 2004
• 期刊: J.Biol.Chem. 279
• 作者: Tamamori-Adachi M.;Hayashida K;Nobori K;Omizu C;Yamada K;Sakamoto N;Kamura T;Fukuda K;Ogawa S;Nakayama KI;Kitajima S
• 通讯作者: Kitajima S

Regulation of hepatitis C virus replication by interferon regulatory factor 1

• DOI: 10.1128/jvi.78.18.9713-9720.2004
• 发表时间: 2004-09-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Kanazawa, N;Kurosaki, M;Watanabe, M
• 通讯作者: Watanabe, M