Systems Biology Identifies Restriction Factors for West Nile Virus

项目概述（见说明）：黄病毒属的成员是最重要的节肢动物传播的病毒，可引起人类疾病，其中许多被美国国立卫生研究院列为A、B和C类生物。该属包括在世界许多地区流行并继续传播的病毒[西尼罗病毒（WNV）、日本脑炎病毒（JEV）和登革热病毒（DENV）]。黄病毒每年造成-1亿人感染，其中数十亿人处于危险之中，而且没有专门的治疗方法。尽管许多工作都集中在了解黄病毒在培养细胞中的复制机制，以及确定体内毒力，但对宿主对不同毒力菌株的反应以及调节体内感染的基因和细胞类型知之甚少。在这里，我们建议鉴定和定义新的宿主基因的作用机制，调节西尼罗河病毒感染。采用系统生物学方法研究与强毒和减毒西尼罗河病毒感染相关的复杂宿主-病原体相互作用，将为宿主反应机制提供新的见解。在几个核心（计算、蛋白质组学、代谢组学和脂质组学、数据管理和资源传播）的支持下，我们将获得宿主对具有不同致病潜力的西尼罗河病毒菌株感染反应的全球图景。这种分析从未对任何黄病毒进行过。这种研究西尼罗河病毒发病机制的方法将直接与项目1关于甲型流感和埃博拉病毒的工作相互作用，以确定限制本文研究的病毒感染的新的共同基因、网络和途径。系统生物学鉴定的靶基因将在细胞中使用异位表达和基因沉默进行验证，这些表型将有助于优先生成新的KO小鼠，以评估西尼罗河病毒感染背景下靶基因在体内的功能。总的来说，我们的研究将为限制西尼罗河病毒和其他病毒感染的细胞过程提供新的见解，从而可能促进开发抑制病毒传播和疾病的治疗剂的新策略。此外，它可能对理解人类的遗传变异有意义，这可以解释对特定病毒感染的易感性。

PROJECT SUMMARY (See instructions): Members of the Flavivirus genus are the most important arthropod-borne viruses causing disease in humans, many of which are characterized as NIH Category A, B, and C organisms. This genus includes viruses [West Nile virus (WNV), Japanese encephalitis virus (JEV) and Dengue virus (DENV)] that are endemic and continue to spread in many areas of the world. Flaviviruses account for -100 millions infections per year, with billions at risk and no specific therapy available. Although much work has focused on understanding the mechanisms of flavivirus replication in cultured cells, and on defining virulence in vivo, less remains known about host responses to strains of different virulence, and the genes and cell types that modulate infection in vivo. Here, we propose to identify and define the mechanism of action of novel host genes that modulate WNV infection. A systems biology approach to studying complex host-pathogen interactions associated with infection by virulent and attenuated WNV strains will provide new insight into host response mechanisms. With the support of several Cores (computational; proteomics, metabolomics, and lipidomics; data management and resources dissemination), we will acquire a global picture of the host response to infection by WNV strains of distinct pathogenic potential. Such an analysis has never been performed for any flavivirus. This approach to studying WNV pathogenesis will interact directly with work in Project 1 on influenza A and Ebola viruses to identify novel common genes, networks, and pathways that restrict infection with the viruses studied here. Target genes identified by systems biology will be validated in cells using ectopic expression and gene silencing, and these phenotypes will help prioritizing the generation of new KO mice for evaluation of the function of target genes in vivo in the context of WNV infection. Overall, our studies will provide new insight into the cellular processes that restrict infection by WNV and likely other viruses, and thus may promote novel strategies for development of therapeutic agents that contain virus spread and disease. Moreover, it may have implications for understanding the genetic variation in humans, which could explain susceptibility to particular viral infections.