Comprehensive screening of host proteins that suppress replication of HCV replicon

全面筛选抑制HCV复制子复制的宿主蛋白

• 批准号: 17590626
• 负责人: SAKAMOTO Naoya
• 金额: $ 2.24万
• 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: 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-17590626/
• 关键词: Hepatitis C virus; HCV replicon system; Cyclosporin A; Interferon-stimulated genes; type-I interferon; high-throughput screening; HCVレプリコン; サイクロフィリン; HCVレプリコン・システム; サイクロスポリンA; 抗ウイルス療法; siRNA; 遺伝子導入療法

We have conducted studies of comprehensive screening of host proteins that suppress hepatitis C virus (HCV) replication using HCV subgenomic replication models and cell culture models.(1) High throughput screening of a library of 2,500 bioactive drugs, peptides, and compounds : we found 52 compounds that suppress or augment HCV replication (Gastroenterology 2006).(2) Establishment of antiviral therapies that target molecular chaperone and cyclophillin : we have reported that cyclosporine A suppress HCV replication through blockade of action of cyclophiline A, B and C. In this study, we next screened host proteins that interact with cyclophillins by bacterial two-hybrid system assay, and identified several proteins including G-protein coupled protein. We are now conducting studies to investigate functions of the proteins.(3) Screening of interferon-stimulated genes that suppress HCV replication : type-I interferon is a key molecule to mediate host virus defense functions. We have newly identified that three IGSs, GBP1, IFI27 and IFI6-16, show suppressive effects on HCV replication. Through immune precipitation assay, we have found that GBP-1 specifically binds HCV-NS5B RNA polymerase.With the above result we are now further conduct screening of antiviral compounds. These results may contribute to establish novel antiviral therapeutics.

我们已经使用HCV亚基因组复制模型和细胞培养模型进行了抑制丙型肝炎病毒（HCV）复制的宿主蛋白质的全面筛选的研究。(1)对2,500种生物活性药物、肽和化合物的文库进行高通量筛选：我们发现了52种抑制或增强HCV复制的化合物（Gastroenterology 2006）。(2)以分子伴侣和亲环蛋白为靶点的抗病毒治疗的建立：我们已经报道了环孢素A通过阻断亲环蛋白A、B和C的作用来抑制HCV复制。在本研究中，我们接下来通过细菌双杂交系统分析筛选与亲环素相互作用的宿主蛋白，并鉴定了包括G蛋白偶联蛋白在内的几种蛋白。我们现在正在研究这些蛋白质的功能。(3)干扰素刺激抑制HCV复制基因的筛选：I型干扰素是介导宿主病毒防御功能的关键分子。我们新发现的三个IGSGBP 1、IFI 27和IFI 6 -16对HCV复制具有抑制作用。通过免疫沉淀实验，我们发现GBP-1与HCV-NS 5 B RNA聚合酶有特异性结合，在此基础上，我们正在进一步进行抗病毒化合物的筛选。这些结果可能有助于建立新的抗病毒治疗方法。

项目成果

肝疾患レビュー 2-I : 肝炎ウイルスと自然免疫

肝病复习2-I：肝炎病毒和先天免疫

• 发表时间: 2006
• 作者: Iwasaki H;Kajimura M;Osawa S;Kanaoka S;Furuta T;Ikuma M;Hishida A.;坂本 直哉(分担);坂本直哉(分担)
• 通讯作者: 坂本直哉(分担)

A cell-based, high-throughput screen for small molecule regulators of HCV replication.

基于细胞的 HCV 复制小分子调节剂高通量筛选。

• 发表时间: 2006
• 期刊: Gastroenterology. 132
• 作者: Kim SS;Sakamoto N;Chung RT et al.
• 通讯作者: Chung RT et al.

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

Enhancement of Mitochondrial Gene Expression in the Liver of Primary Biliary Cirrhosis.

原发性胆汁性肝硬化肝脏中线粒体基因表达的增强。

• 发表时间: 2005
• 期刊: Hepatology Res. 31
• 作者: Chen CH;Nagayama K;Sakamoto N;Watanabe M;et al.
• 通讯作者: et al.

Mutations in the NS5B region of the hepatitis C virus genome correlate with clinical outcomes of interferon-alpha plus ribavirin combination therapy

• DOI: 10.1111/j.1440-1746.2005.04024.x
• 发表时间: 2005-09-01
• 期刊: JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY
• 影响因子: 4.1
• 作者: Hamano, K;Sakamoto, N;Watanabe, M
• 通讯作者: Watanabe, M