Characterization of a host hepatocyte factor that regulates HCV replication

描述（由申请人提供）：慢性丙型肝炎病毒（HCV）感染发生在所有暴露个体的70-80%中，影响世界人口的3%。需要更有效和耐受性更好的丙肝病毒治疗方法。我们的长期目标是确定宿主-丙型肝炎病毒分子相互作用，并在此过程中确定丙型肝炎病毒治疗的新药物。我们已经使用全基因组siRNA文库完成了HCV复制宿主辅助因子的功能基因组筛选。在许多新的宿主辅助因子中，PI4KA是一种磷脂酰肌醇4-激酶（pi4 -激酶）。我们假设PI4KA对于HCV复制所需的改变宿主膜的形成是必要的。该提案描述了一个为期5年的指导培训计划，以获得分子病毒学，磷酸肌苷和细胞运输以及细胞成像领域的进一步技术技能。这些技能将用于开发新的模型系统，以研究HCV复制复合体及其相关宿主膜和蛋白质的形成和组成。该项目的具体目标是：(1)表征PI4KA抑制HCV复制的特异性，(2)表征PI4KA在病毒复制周期中的作用，(3)开发研究HCV复制复合体组装和组成的系统。虽然具体目标是针对PI4KA，但在奖励期间开发的系统和技术将应用于HCV复制的其他宿主辅因子的研究。拟议的研究将有直接的公共卫生效益，因为更好地理解强调HCV复制复合体组装的机制可能导致HCV治疗的新靶点。更好地了解hcv -宿主关系是NIH肝病研究行动计划（目标B2a）的既定目标。我们最近对丙型肝炎病毒复制所需的人类（宿主）蛋白质进行了全基因组筛选。这些蛋白是HCV新疗法的潜在靶点。该提案描述了一个5年的研究计划，重点是开发新的系统来研究宿主辅助因子如何指导HCV复制复合体的组装。

DESCRIPTION (provided by applicant): Chronic hepatitis C virus (HCV) infection develops in 70-80% of all exposed individuals and affects 3% of the world's population. There is a need for more effective and better-tolerated HCV therapies. Our long-term goals are to define host-HCV molecular interactions and in so doing, to identify novel therapeutic agents for HCV therapy. We have completed a functional genomic screen for host cofactors for HCV replication using a whole- genome siRNA library. Among the many novel host cofactors identified is PI4KA, a phosphatidylinositol 4- kinase (PI 4-kinase). We hypothesize that PI4KA is necessary for formation of the altered host membranes required for HCV replication. This proposal describes a 5 year mentored training program to acquire further technical skills in the fields of molecular virology, phosphoinositides and cell trafficking, and cell imaging. These skills will be used to develop novel model systems to study the formation and composition of the HCV replication complex and its associated host membranes and proteins. The specific aims of this project are (1) to characterize the specificity of PI4KA inhibition for suppression of HCV replication, (2) to characterize the role of PI4KA through the viral replication cycle, and (3) to develop systems to study HCV replication complex assembly and composition. Although the specific aims are directed towards PI4KA, the systems and techniques to be developed during the award period will be applied to the study of other host cofactors of HCV replication. The proposed research will have direct public health benefits as a better understanding of the mechanisms that underline HCV replication complex assembly may lead to novel targets for HCV therapies. A better understanding of the HCV-host relationship is a stated goal of the NIH Action Plan for Liver Disease Research (Goal B2a). We have recently performed a whole-genome screen for human (host) proteins that are required by the hepatitis C virus for its replication. Such proteins are potential targets for new therapies for HCV. This proposal describes a 5-year research plan focusing on developing new systems to study how host cofactors direct the assembly of HCV replication complexes.