Studies of hepatitis C virus polarized cell entry and host factor requirements

丙型肝炎病毒（HCV）是美国肝脏疾病的主要原因。无特异性抗HCV 虽然干扰素治疗是一种有效的治疗方法，但目前采用的基于干扰素的治疗是不够的，因为它具有严重的副作用， 仅对一半的主要基因型感染个体有效。本申请中提出的研究 旨在进一步了解HCV的复制机制，最终目标是。. 发现新的抗病毒靶点。在过去，我研究了HCV的复制是如何被调节的，病毒是如何 蛋白质相互作用和细胞因子，并帮助建立系统，研究丙型肝炎病毒在细胞中， 文化目前，我的研究主要集中在病毒生命周期中最早的事件，包括进入 宿主细胞，这是知之甚少的过程。特别是，尽管已经有几个主机因素 与HCV进入有关，|我们知道病毒如何利用它们。我们最近 紧密连接蛋白Claudin-1（CLDN 1）的鉴定是该领域的一个主要进展。这一发现开辟了HCV细胞进入的全新视角，因为CLDN 1参与细胞极性强烈表明肝细胞的极化性质可能影响HCV如何进入细胞。该提案描述了以下实验：1）研究HCV进入极化细胞，2）定义病毒体如何与已知的HCV进入因子相互作用并利用已知的HCV进入因子，3）对能够使人和鼠细胞都感染HCV的HCV进入因子进行额外的筛选。 这样的研究将提供更大的，急需的深入了解丙型肝炎病毒的复制，以及奠定 为今后研究HCV和其他相关病毒的复制奠定了基础。

Hepatitis C virus (HCV) is the leading cause of liver disease in the United States. With no specific anti-HCV therapies, the cun-ently employed interferon-based treatment is inadequate, as it has severe side effects and is only effective in half of the major genotype infected individuals. The research proposed in this application is aimed at furthering the understanding the HCV replication mechanisms, with the ultimate goal of . . uncovering novel antiviral targets. In the past, I have studied how HCV replication is regulated, how viral proteins interact with each other and cellular factors, and helped establish systems to study HCV in cell culture. Currently, my research is focused on the earliest events in the viral life cycle involving entry into the host cell, which are poorly understood processes. In particular, although several host factors have been implicated as involved in HCV entry, litt|e is know as to how they are utilized by the virus. Our recent identification ofthe tight junction protein, claudin-1 (CLDN1), as essential for this process was a major development in this field. This discovery has opened-up a whole new view of HCV cell entry, as the involvement of CLDN1 in cell polarity strongly suggests that the polarized nature of an hepatocyte may influence how HCV enter a cell. This proposal describes experiments to; 1) study HCV entry into polarized cells, 2) define how the virion interacts with and utilizes the known HCV entry factors, and 3) perform additional screens for HCV entry factors able to render both human and murine cells infectable with HCV. Such investigations will provide greater, much needed insight in HCV replication, as well as lay the foundation for future studies of replication of HCV and other related viruses.