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

丙型肝炎病毒(丙型肝炎病毒)是美国肝病的主要原因。没有特定的抗丙型肝炎病毒抗体 在治疗方面，目前使用的以干扰素为基础的治疗是不够的，因为它有严重的副作用和 只对一半的主要基因感染个体有效。在本申请中提出的研究 旨在加深对丙型肝炎病毒复制机制的了解，最终目标是。。 发现新的抗病毒靶点。过去，我曾研究过丙型肝炎病毒复制是如何调控的，病毒是如何传播的 蛋白质与细胞因子相互作用，并帮助建立了在细胞中研究丙型肝炎病毒的系统 文化。目前，我的研究重点是病毒生命周期中最早的事件，涉及进入 宿主细胞，这是知之甚少的过程。特别是，尽管几个主机因素已经被 由于牵涉到了丙型肝炎病毒的进入，对病毒如何利用它们知之甚少。我们最近 紧密连接蛋白Claudin-1(CLDN1)的鉴定是该领域的一项重大进展。这一发现开启了对丙型肝炎病毒细胞进入的全新看法，因为CLDN1参与细胞极性的研究强烈表明，肝细胞的极化性质可能会影响丙型肝炎病毒进入细胞的方式。该建议描述了以下实验：1)研究丙型肝炎病毒进入极化细胞，2)确定病毒粒子如何与已知的丙型肝炎病毒进入因子相互作用和利用，以及3)对能够使人类和小鼠细胞感染丙型肝炎病毒的丙型肝炎病毒进入因子进行额外的筛选。 这样的研究将为丙型肝炎病毒复制提供更多、更必要的洞察力，并为 为未来研究丙型肝炎病毒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.