Identification of Host Drug Development Targets in Influenza Using Transposon Mutagenesis

使用转座子诱变鉴定流感宿主药物开发靶点

• 批准号: 8956296
• 负责人: Adam Lacy-Hulbert
• 金额: $ 21.75万
• 依托单位: BENAROYA RESEARCH INST AT VIRGINIA MASON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8956296
• 关键词: Animals; Antiviral Agents; Antiviral resistance; Bioinformatics; Candidate Disease Gene; Cell Death; Cells; Cessation of life; Data; Drug Targeting; Drug resistance; Ebola virus; Elements; Epidemic; Gene Expression; Generations; Genes; Genetic; Goals; Healthcare; High-Throughput Nucleotide Sequencing; Host Defense; Host Defense Mechanism; Human; Human Cell Line; Immune system; Immunity; Individual; Infection; Infection prevention; Influenza; Influenza A virus; Integration Host Factors; Laboratories; Lead; Libraries; Mediating; Methodology; MicroRNAs; Modeling; Morbidity - disease rate; Mutagenesis; Mutation; Outcome; Pathway interactions; Phase; Physiological; Public Health; RNA Interference; Resistance; Resistance to infection; Site; Testing; Therapeutic; Therapeutic Intervention; Transcript; Untranslated RNA; Up-Regulation; Vaccination; Vesicular stomatitis Indiana virus; Viral; Virus; Virus Diseases; Work; base; care burden; cell type; chemotherapy; combat; drug development; drug mechanism; gene interaction; genetic approach; genome-wide; influenzavirus; innovation; insight; mortality; mutant; new therapeutic target; novel; overexpression; pandemic disease; prevent; programs; public health relevance; resistance gene; resistant strain; screening; seasonal influenza; small molecule; success; targeted treatment; therapeutic target; viral resistance

 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.

 描述（由申请人提供）：季节性流感持续的医疗负担、对现有治疗产生耐药性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防御感染的机制。本应用的目的是确定宿主编码的抵抗流感感染的机制。根据新出现的数据和我们的初步研究，我们的中心假设是，阻止病毒进入的宿主“限制因子”的上调限制了病毒在宿主细胞中感染或复制的能力，或增加了细胞抵抗病毒诱导的细胞病变的能力，代表了宿主防御的重要策略。我们提出的工作的基本原理是，识别这些因素将为治疗干预提供大量新靶标，并且这些靶标可能比病毒编码靶标更不易受到耐药性的影响。在 R21 阶段，我们建议使用一种新颖的无偏正向遗传方法来筛选能够抵抗流感感染的宿主基因。在 R33 阶段，我们将验证新的宿主靶标，并利用这些见解来开发新的抗病毒治疗策略。我们的筛选方法是创新的，因为它可以识别过度表达时赋予抗性的宿主基因（“限制因子”）以及感染所需的宿主基因，这些基因通常在传统的基于 RNAi 的筛选中检测到。拟议的工作意义重大，因为它有可能识别新的靶标，包括非编码 RNA 元件，而使用现有方法可能会错过这些靶标。此外，我们的方法对 RFA 做出响应，因为它旨在确定新的宿主靶点和治疗策略，以对抗甲型流感并克服对现有抗病毒药物的耐药性。