The role of interferon-stimulated genes in antiviral immunity

描述（由申请方提供）：干扰素（IFN）是一种抗病毒细胞因子，通过诱导300+干扰素刺激基因（ISG）介导其活性。这些基因中有几个是众所周知的病毒复制抑制剂，但大多数ISG的抗病毒潜力仍不确定。微阵列和基因敲除研究表明，许多这些ISG具有显着的抗病毒功能。本项目的长期目标是鉴定和表征对丙型肝炎病毒（HCV）（非甲非乙型病毒性肝炎的病原体）表现出新型抗病毒活性的ISG。我们的实验方法依赖于产生一系列表达所有已知ISG的基于慢病毒的载体。将在高通量试验中筛选收集的样本，以确定过度表达时抑制HCV复制的基因。我们将利用最近开发的HCV感染的细胞培养模型来进行这种筛选。候选抗HCV ISG将通过功能测定进一步表征，以确定其作用机制。我们还计划利用我们的基于筛选的方法来鉴定对不同病毒家族表现出新的抗病毒活性的ISG。我们的分析将包括几种重要的人类病原体，包括登革热病毒（DV）和甲型流感病毒（FluA）。来自每个病毒家族的代表性成员的数据将被组装成“ISG谱”，该谱将用于跨家族比较分析。这项研究的重点是，虽然病毒是人类疾病的重要贡献者，但IFN等抗病毒疗法通常无效且有毒。HCV影响世界人口的约3%，并显著导致肝硬化和肝癌; DV是一个全球性的健康问题，感染数百万人并引起登革热和严重的登革出血热;仅在美国，流感每年就导致超过30，000人死亡。鉴定对这些和其他病毒表现出抗病毒活性的基因对于产生新型高效抗病毒治疗剂是重要的。

DESCRIPTION (provided by applicant): Interferon (IFN) is an antiviral cytokine that mediates its activity through the induction of 300+ interferon-stimulated genes (ISGs). Several of these genes are well-known inhibitors of virus replication, but the majority of the ISGs remain uncharacterized with respect to their antiviral potential. Microarray and knock-out studies suggest that many of these ISGs possess significant antiviral functions. The long-term objectives of this project are to identify and characterize ISGs that exhibit novel antiviral activity in response to hepatitis C virus (HCV), the etiological agent of non-A, non-B viral hepatitis. Our experimental approach relies on generating a collection of lentiviral-based vectors that express all known ISGs. The collection will be screened in a high-throughput assay for genes that inhibit HCV replication when overexpressed. We will take advantage of the recently developed cell culture model of HCV infection to carry out this screen. Candidate anti-HCV ISGs will be further characterized with functional assays to determine their mechanisms of action. We also plan to utilize our screen-based approach to identify ISGs that exhibit novel antiviral activity against different virus families. Several significant human pathogens, including Dengue virus (DV) and influenza A virus (FluA), will be included in our analysis. Data from representative members of each virus family will be assembled into an "ISG profile," which will be used for cross-family comparative analysis. This research is highlighted by the fact that although viruses are significant contributors to human disease, antiviral therapies such as IFN are often ineffective and toxic. HCV affects approximately 3 percent of the world's population and contributes significantly to cirrhosis and liver cancer; DV is a global health concern that infects millions of people and causes dengue fever and the severe dengue hemorrhagic fever; FluA results in the death of more than 30,000 people each year in the US alone. Identifying genes that exhibit antiviral activity against these and other viruses is important for generating novel, highly effective antiviral therapeutic agents.