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

描述（由申请人提供）：目前慢性丙型肝炎病毒（HCV）感染的治疗包括干扰素（IFN）和抗病毒前药利巴铁的联合治疗。干扰素是先天免疫系统中一个具有良好特征的组成部分，它通过干扰素刺激基因（ISGs）的活性抑制多种病毒。令人惊讶的是，大多数已知的isg的特征很差，它们的抗病毒作用机制也不明确。先前实验室进行的抗病毒ISG筛选揭示了细胞蛋白Mov10在HCV抑制中的作用。通过各种病毒学、生化和计算方法确定HCV生命周期中的哪些阶段受到影响，将进一步确定Mov10的抗病毒活性。这项工作将阐明Mov10在病毒感染中的作用，并增加我们目前对病毒-宿主相互作用的理解。此外，由于缺乏对已知ISG的机制理解，我们仍然缺乏ISG诱导网络的层次视图。干扰素的刺激导致isg复杂网络的建立，该网络可能由反馈回路和阈值效应控制。虽然已知许多基因对IFN有反应，但对isg的复杂网络是如何建立和控制的知之甚少。我们建议通过使用RNA-seq方法以剂量和时间依赖的方式对IFN处理的细胞进行转录组分析来解决这一科学知识上的差距。在本研究中，原代人肝细胞（HCV的天然储存库）将重点用于低干扰素浓度的实验。

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.