Identification of Host Drug Development Targets in Influenza Using Transposon Mutagenesis

 描述（由申请方提供）：季节性流感的持续医疗负担、对现有治疗方法耐药的菌株的出现以及新的大流行菌株的持续威胁，突出表明迫切需要确定针对流感的新治疗靶点。我们实验室的长期目标是确定宿主对感染的防御机制。本申请的目的是鉴定宿主编码的抗流感感染的机制。基于新出现的数据和我们的初步研究，我们的中心假设是，上调宿主的“限制因子”，防止病毒进入限制病毒感染或复制的能力，在宿主细胞或增加的能力，抵御病毒诱导的细胞病变的宿主防御代表一个重要的战略。我们提出这项工作的理由是，这些因素的鉴定将为治疗干预提供丰富的新靶点，并且这些靶点可能比病毒编码的靶点更不易受到耐药性的影响。在R21阶段，我们建议使用一种新的无偏正向遗传方法来筛选赋予对流感感染的抗性的宿主基因。在R33阶段，我们将验证新的宿主靶点，并利用这些见解开发新的抗病毒治疗策略。我们的筛选方法是创新的，因为它允许鉴定宿主基因，当过度表达时赋予抗性（“限制因子”）以及感染所需的宿主基因，这通常在传统的基于RNAi的筛选中检测到。拟议的工作是重要的，因为它有可能确定新的目标，包括非编码RNA元件，这将是错过使用现有的方法。此外，我们的方法是响应RFA，因为它是针对确定新的主机目标和治疗策略，以打击甲型流感和克服现有的抗病毒药物的耐药性。

 DESCRIPTION (provided by applicant): The ongoing healthcare burden of seasonal Influenza, the emergence of strains resistant to existing treatments and the constant threat of new pandemic strains highlight the urgent need to identify new therapeutic targets against Influenza. The long term goal of our laboratory is to identify mechanisms of host defense against infection. The objective in this application is to identify host-encoded mechanisms of resistance to infection by Influenza. Based on emerging data and our preliminary studies, our central hypothesis is that the up-regulation of host 'restriction factors' that prevent viral entry restrict the ability of viruses to infect or replicate in host cells or increase the ability of cels to withstand viral-induced cytopathy represent an important strategy in host defense. Our rationale for the proposed work is that identification of such factors will provide a wealth of new targets fr therapeutic intervention, and that these may be less susceptible to resistance than viral-encoded targets. In the R21 phase we propose to use a novel unbiased forward-genetic approach to screen for host genes that confer resistance to Influenza infection. In the R33 phase, we will validate new host targets and use these insights to develop new anti-viral therapeutic strategies. Our screening approach is innovative because it allows identification of both host genes that, when overexpressed confer resistance ('restriction factors') as well as host genes required for infection, which are normally detected in conventional RNAi-based screens. The proposed work is significant because it has the potential to identify new targets, including non- coding RNA elements, which would be missed using existing approaches. Furthermore, our approach is responsive to the RFA because it is directed at identifying new host targets and therapeutic strategies to combat Influenza A and overcome resistance to existing antiviral agents.