Systems Biology Identifies Restriction Factors for West Nile Virus

项目总结（见说明）：黄病毒属成员是引起人类疾病的最重要的节肢动物传播病毒，其中许多被鉴定为NIH A类、B类和C类微生物。该属包括病毒[西尼罗河病毒（WNV），日本脑炎病毒（JEV）和登革热病毒（DENV）]，这些病毒是地方性的，并继续在世界许多地区传播。黄病毒每年造成约1亿例感染，数十亿人处于风险中，并且没有可用的特异性治疗。虽然很多工作都集中在了解黄病毒在培养细胞中复制的机制，并在体内定义毒力，仍然知道较少的宿主反应不同毒力的菌株，以及基因和细胞类型，调节体内感染。在这里，我们建议确定和定义新的宿主基因，调节西尼罗河病毒感染的作用机制。一个系统生物学的方法来研究复杂的宿主-病原体相互作用与感染的毒力和减毒西尼罗河病毒株将提供新的见解宿主反应机制。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株感染的反应的全球图景。从未对任何黄病毒进行过此类分析。这种研究西尼罗河病毒发病机制的方法将直接与项目1中关于甲型流感和埃博拉病毒的工作相互作用，以确定限制感染本文研究的病毒的新的共同基因、网络和途径。通过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产生新的KO小鼠，用于在WNV感染的背景下评估体内靶基因的功能。总的来说，我们的研究将为限制WNV和可能的其他病毒感染的细胞过程提供新的见解，从而可能促进开发包含病毒传播和疾病的治疗药物的新策略。此外，它可能对理解人类的遗传变异有影响，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.