Regulation of host cell gene expression by the hepatitis C virus protein NS5A

丙型肝炎病毒蛋白 NS5A 对宿主细胞基因表达的调节

• 批准号: 8400739
• 负责人: Jose Daniel Debes
• 金额: $ 6.1万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8400739
• 关键词: 3' Untranslated Regions; Adverse effects; Affect; Antibodies; Binding; Cell Death; Cell Line; Cell Proliferation; Cells; Cessation of life; Chronic; Cirrhosis; Data; Dissection; Down-Regulation; Elements; Exhibits; Gene Expression; Gene Targeting; Genes; Growth; Hepatitis C; Hepatitis C virus; Human Cell Line; Immunoprecipitation; Indium; Infection; Laboratories; Liver Cirrhosis; Liver Fibrosis; Liver diseases; Malignant Neoplasms; Mediating; Messenger RNA; Nonstructural Protein; Northern Blotting; Oligonucleotide Microarrays; Pharmaceutical Preparations; Play; Primary carcinoma of the liver cells; Protein Binding; Proteins; Proto-Oncogenes; RNA; Regulation; Reporter; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Small Interfering RNA; Spottings; Transcript; Viral; Viral Proteins; Virus; Virus Inhibitors; Virus Replication; hepatoma cell; inhibitor/antagonist; innovation; knock-down; mRNA Decay; mRNA Transcript Degradation; mortality; novel; overexpression; particle; prevent; response; standard care

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) is a major cause of liver disease and mortality, affecting approximately 170 million people worldwide. Chronic infection with HCV leads to liver fibrosis, cirrhosis and hepatocellular carcinoma. HCV must use the host gene expression machinery to facilitate viral replication. The mechanisms involved in the replication are incompletely understood. It has been found that the HCV nonstructural protein NS5A is essential to the replication machinery of the virus and critical in the assembly of infectious viral particles, however its specific role remains unclear. Recent preliminary data from the Bohjanen's laboratory indicates that the HCV NS5A protein binds to host GRE-containing transcripts and mediates their stabilization in hepatoma cell lines. This data suggest that HCV could manipulate host cellular mRNA decay through NS5A- mediated stabilization of GRE-containing transcripts. Stabilization of these transcripts would prevent cell death and promote growth of virus -infected cells allowing the virus to establish chronic infection in the host. In this study, we aim to identify host cellular transcripts that bind to the hepatitis C virus non-structural protein and to determine if the NS5A inhibitor BMS 790052 blocks HCV replication through interference with the binding of NS5A to GRE-containing host cellular transcripts. Further understanding of the critical genes involved in RNA regulation of HCV in human cell lines, will help characterize the virus replication cycle. In addition, it will expose genes of critical importance, thus unveiling potential targets for treatment. Moreover, dissection of the downstream effects of NS5A inhibitors will expose additional weak spots in the HCV replication machinery, expanding our understanding of the virus and facilitating innovation of new therapies.

描述（由申请人提供）：丙型肝炎病毒 (HCV) 是肝病和死亡的主要原因，影响全球约 1.7 亿人。慢性感染HCV会导致肝纤维化、肝硬化和肝细胞癌。 HCV必须利用宿主基因表达机制来促进病毒复制。复制所涉及的机制尚不完全清楚。研究发现，HCV 非结构蛋白 NS5A 对于病毒的复制机制至关重要，并且在感染性病毒颗粒的组装中至关重要，但其具体作用仍不清楚。 Bohjanen 实验室最近的初步数据表明，HCV NS5A 蛋白与宿主含有 GRE 的转录物结合，并介导其在肝癌细胞系中的稳定性。该数据表明，HCV 可以通过 NS5A 介导的含有 GRE 转录物的稳定性来操纵宿主细胞 mRNA 的衰变。这些转录物的稳定将防止细胞死亡并促进病毒感染细胞的生长，从而使病毒在宿主体内建立慢性感染。在这项研究中，我们的目的是鉴定与丙型肝炎病毒非结构蛋白结合的宿主细胞转录本，并确定 NS5A 抑制剂 BMS 790052 是否通过干扰 NS5A 与含有 GRE 的宿主细胞转录本的结合来阻断 HCV 复制。进一步了解人类细胞系中 HCV RNA 调节所涉及的关键基因，将有助于表征病毒复制周期。此外，它将暴露至关重要的基因，从而揭示潜在的治疗目标。此外，对 NS5A 抑制剂下游效应的剖析将暴露 HCV 复制机制中的其他弱点，扩大我们对病毒的了解并促进新疗法的创新。