Hepatitis C virus and the innate immune response: from transcriptome to function

描述(申请人提供)：目前慢性丙型肝炎病毒(丙型肝炎病毒)感染的治疗包括干扰素(干扰素)和抗病毒前药利巴韦隆的联合治疗。干扰素是先天免疫系统的一个重要组成部分，它通过干扰素刺激基因(ISGs)的活动抑制多种病毒。令人惊讶的是，大多数已知的ISG的特性很差，其抗病毒作用的机制也不明确。实验室先前的抗病毒ISG筛查发现了细胞蛋白Mov10在抑制丙型肝炎病毒中的作用。Mov10将通过各种病毒学、生化和计算方法确定丙型肝炎病毒生命周期的哪些阶段受到影响，从而进一步定义它们的抗病毒活性。这项工作将阐明Mov10在病毒感染中的作用，并加深我们目前对病毒与宿主相互作用的理解。此外，由于缺乏对已知ISG的机械性理解，我们仍然缺乏对ISG感应网络的分层观点。干扰素的刺激导致建立一个复杂的ISGs网络，该网络可能受反馈环和阈值效应的控制。虽然已知有许多基因对干扰素有反应，但关于ISGs的复杂网络是如何建立和控制的，人们知之甚少。我们建议通过使用RNA-SEQ方法，以剂量和时间依赖的方式对干扰素处理的细胞进行转录组分析，以解决科学知识中的这一缺口。在这项建议中，原代人肝细胞，丙型肝炎病毒的天然储存库，将被用于强调低浓度的干扰素。

DESCRIPTION (provided by applicant): Current therapy for chronic hepatitis C virus (HCV) infection consists of treatment with a combination of interferon (IFN) and the antiviral prodrug ribaviron. IFNs are a well-characterized component of the innate immune system that inhibits a wide range of viruses through the activities of interferon-stimulated genes (ISGs). Surprisingly, the majority of known ISGs are poorly characterized and their mechanism of antiviral action undefined. A previous antiviral ISG screen with the lab uncovered a role of the cellular protein Mov10 in HCV inhibition. Mov10 will be further defined with respect to their antiviral activity by identifying what stages in the HCV lifecycle are affected through various virological, biochemical and computational approaches. This work will clarify the role of Mov10 in viral infection and add to our current understanding of virus-host interactions. In addition a lack in mechanistic understanding of known ISGs we continue to lack a hierarchal view of the ISG induction network. Stimulation by IFN results in the establishment of a complex network of ISGs that is likely controlled by feedback loops and threshold effects. While many genes are known to respond to IFN, little is known of how the complex network of ISGs is established and controlled. We propose to address this gap in scientific knowledge by performing transcriptome analysis on cells treated with IFN in a dose- and time-dependent manner using the RNA-seq method. In this proposal, primary human hepatocytes, the natural reservoirs for HCV, will be used with an emphasis on low IFN concentrations.