Molecular determinants of hepatitis C virus infectivity

描述（由申请人提供）：丙型肝炎病毒（HCV）是一种非常成功的病原体，可导致慢性肝炎和肝细胞癌。目前用于治疗HCV患者的疗法通常无效，并且尚未获得HCV特异性抗病毒药物和疫苗。针对这种病原体的新机会可能来自对病毒生命周期的彻底了解。我们最近开发了一种在细胞培养中有效生长HCV的系统，使病毒的生命周期在分子水平上得到充分剖析。我们将使用这个系统来研究传染性病毒颗粒是如何组装的。一个关键的观察结果是，感染性HCV颗粒具有异常低的浮力密度，这可能是由于与血清脂蛋白的相互作用。在这个建议中，我们研究的假设，HCV颗粒的感染性需要在病毒形成的细胞内步骤与血清脂蛋白成分的相互作用。为了解决这一假设，我们将定义HCV颗粒的组成（目标1）;通过使用活细胞和固定细胞成像方法识别HCV颗粒组装的早期步骤（目标2）;并通过使用生物化学和细胞生物学方法描绘HCV颗粒成熟的细胞内过程（目标3）。这些研究将为HCV颗粒感染性和病毒-宿主相互作用的本质提供新的见解。公共卫生相关性：丙型肝炎病毒（HCV）是一种非常成功的人类病原体，可引起持续感染、慢性肝炎和肝细胞癌。我们的研究特征的病毒颗粒及其与宿主细胞的相互作用，这将提供新的途径，以针对HCV。

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) is a highly successful pathogen that causes chronic hepatitis and hepatocellular carcinoma. Current therapies for treating HCV patients are frequently ineffective, and HCV-specific antivirals and vaccines are not yet available. New opportunities for targeting this pathogen are likely to come from a thorough understanding of the viral life cycle. We have recently developed a system to efficiently grow HCV in cell culture, allowing the viral life cycle to be fully dissected at the molecular level. We will use this system to examine how infectious virus particles are assembled. One key observation is that infectious HCV particles have an unusually low buoyant density, which is likely due to interaction with serum lipoproteins. In this proposal, we examine the hypothesis that HCV particle infectivity requires interaction with serum lipoprotein components during an intracellular step in virus formation. To address this hypothesis, we will define the composition of HCV particles (Aim 1); identify early steps in HCV particle assembly by using live and fixed cells imaging methods (Aim 2); and delineate the intracellular process of HCV particle maturation by using biochemical and cell biological methods (Aim 3). These studies will provide new insights into the nature of HCV particle infectivity and virus-host interaction. PUBLIC HEALTH RELEVANCE: Hepatitis C virus (HCV) is a highly successful human pathogen that causes persistent infection, chronic hepatitis, and hepatocellular carcinoma. Our studies characterize the virus particle and its interactions with the host cell, which will provide new avenues to target HCV.