Molecular components specifying mucosal cell sensitivity to influenza infection

描述（由申请人提供）：通过对细胞自主分子成分的更广泛理解，将极大地促进转移或改善流感感染的新策略的发展，这些分子成分被颠覆以支持病毒感染，以及那些被动员起来抵御病毒感染的分子成分。全基因组siRNA文库的出现，加上单孔/单基因高通量筛选策略，为生成实验平台提供了机会，以获得支持关键生物系统的经过验证的基因靶点的公正综合集合。我们的方法是应用基于全基因组rnai的体细胞遗传学对人类支气管上皮细胞中的基因产物进行广泛和公正的鉴定，这些基因产物支持1)流感生命周期和2)抗病毒监测和反应系统。在Specific Aim 1中，将利用全基因组siRNA筛选来鉴定暴露于流感a /WS/33后导致（电阻位点）或（致敏位点）支气管上皮细胞死亡的所有基因缺失。在Specific Aim 2中，将采用集中的分层实验策略将验证命中解析为代表关键生物过程的功能互补组。由此产生的控制流感复制和宿主先天免疫途径激活的关键生物压力点的分子注释将有助于1)对宿主/病毒关系产生新的机制见解，2)扩大对宿主系统中支持病毒复制的可靶向生物压力点的认识，以及3)促进识别具有药物样特性的新型生物活性化合物的分子靶标。流感病毒必须劫持存在于人粘膜细胞内的蛋白质，以便在受感染个体中繁殖。详细了解支持病毒复制所需的粘膜细胞蛋白，以及那些被动员起来防御病毒复制的粘膜细胞蛋白，将有助于开发新的治疗和预防干预措施。这里描述的工作将有助于产生一个全面的知识基础的粘膜细胞蛋白，既支持和对抗甲型流感感染和复制。

DESCRIPTION (provided by applicant): Development of novel strategies to deflect or ameliorate influenza infection will be greatly facilitated by a broader understanding of the cell-autonomous molecular components that are subverted to support viral infection, and those that are mobilized to defend against it. The advent of genome-wide siRNA libraries, coupled with one-well/one-gene high throughput screening strategies, provides the opportunity to generate experimental platforms to derive unbiased comprehensive collections of validated gene targets that support critical biological systems. Our approach is to apply genome-wide RNAi-based somatic cell genetics for an extensive and unbiased identification of gene products in human bronchial epithelial cells that support 1) the influenza life cycle and 2) the antiviral surveillance and response system. In Specific Aim 1, a genome-wide siRNA screen will be leveraged to identify all gene depletions that deflect (resistor loci) or promote (sensitizor loci) bronchial epithelial cell death upon exposure to influenza A/WS/33. In Specific Aim 2 a focused hierarchical experimental strategy will be employed to parse validated hits into functional complementation groups representing key biological processes. The resulting molecular annotation of critical biological pressure points controlling flu replication and deflection of host innate immune pathway activation will be used to help 1) generate novel mechanistic insights into the host/virus relationship, 2) broaden knowledge of targetable biological pressure points in host systems that support viral replication, and 3) facilitate identification of the molecular targets of novel bioactive compounds with drug-like properties. Influenza virus must hijack proteins present within the cells of the human mucosa in order to reproduce in infected individuals. A detailed understanding of the mucosal cell proteins required to support viral replication as well as those which are mobilized to defend against it will aid development of novel therapeutic and preventative interventions. The work described here will help produce a comprehensive knowledge base of the mucosal cell proteins that both support and antagonize influenza A infection and replication.