Identification of Host Directed Drug Targets for SARS-CoV-2 Using Transposon Mutagenesis

使用转座子诱变鉴定 SARS-CoV-2 宿主定向药物靶点

基本信息

批准号：
10238213
负责人：
Anna Bruchez
金额：
$ 44.28万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-06 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10238213
关键词：
2019-nCoV Address Antiviral Agents Bioinformatics COVID-19 pandemic Candidate Disease Gene Cell Death Cell Line Cells Coronavirus Country Data Development Disease Outbreaks Drug Targeting Ebola Elements Equilibrium Future Genes Genetic Genetic Screening Goals Health system High-Throughput Nucleotide Sequencing Host Defense Host Defense Mechanism Infection Infection prevention Influenza A virus Innate Immune System Integration Host Factors Laboratories Libraries Mediating Methodology Methods MicroRNAs Mutagenesis Nature Pathogenicity Pathway interactions Public Health RNA interference screen Resistance Resistance to infection SARS-CoV-2 infection Scientist Site Speed Therapeutic Therapeutic Intervention Transcript Untranslated RNA Up-Regulation Vesicular stomatitis Indiana virus Viral Virus Virus Diseases Virus Replication Work antiviral immunity base causal variant cell type falls gene interaction genetic approach genome-wide inhibitor/antagonist innovation insight mutant new therapeutic target novel novel therapeutics overexpression pandemic disease physiologic model prevent public health emergency resistance gene resistance mechanism screening spillover event targeted treatment therapeutic candidate therapeutic target viral resistance virus host interaction

项目摘要

SUMMARY The ongoing SARS-CoV-2 pandemic has brought into stark relief the need for novel therapeutics that can work against a broad spectrum of coronaviruses. Identifying essential host genes involved in both virus replication and antiviral defense could reveal key host-directed viral therapies that have the potential to work against SARS-CoV-2 and future coronavirus outbreaks. The overarching goal of our laboratory is to identify mechanisms of host defense against viral infections. This application aims to identify both host-encoded mechanisms of resistance to infection by SARS-CoV-2 as well as cellular factors critical for virus replication. Based on emerging data and our preliminary studies, our central hypothesis is that the up-regulation of host ‘restriction factors’ that prevent viral entry, constrain virus replication in host cells, or increase the ability of cells 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 new targets for therapeutic intervention, and that these may be less susceptible to resistance than viral-encoded targets. We will use a novel forward-genetic approach to screen for host genes that confer resistance to SARS-CoV-2 infection in BSL3 using the native virus. This will allow us to identify key targets at all stages of virus infection. We will then validate new host targets and use these insights to rationally develop antiviral therapeutic strategies. Our screening approach is innovative because it allows identification of both host genes that confer resistance as well as host genes required for infection, which are normally detected in conventional RNAi-based screens. This work has the potential to identify new targets, including non-coding RNA elements, which would be missed using existing approaches.

概括持续不断的 SARS-CoV-2 大流行凸显了对有效治疗小说的需求针对广泛的冠状病毒，识别参与两种病毒复制的重要宿主基因。抗病毒防御可以揭示关键的宿主导向病毒疗法，这些疗法有可能对抗病毒我们实验室的首要目标是识别 SARS-CoV-2 和未来的冠状病毒爆发。宿主防御病毒感染的机制该应用旨在识别宿主编码的两种机制。抵抗 SARS-CoV-2 感染的机制以及对病毒复制至关重要的细胞因素。根据新出现的数据和我们的初步研究，我们的中心假设是宿主的上调阻止病毒进入、限制病毒在宿主细胞中复制或增加细胞能力的“限制因素” 抵抗病毒诱导的细胞病变是宿主防御的一个重要策略。拟议的工作是，识别这些因素将为治疗干预提供新的目标，并且这些可能比病毒编码的靶标更不容易受到耐药性的影响，我们将使用一种新型的前向遗传。使用天然方法在 BSL3 中筛选赋予对 SARS-CoV-2 感染抵抗力的宿主基因这将使我们能够识别病毒感染各个阶段的关键目标，然后我们将验证新宿主。目标并利用这些见解来合理制定抗病毒治疗策略。创新，因为它可以识别赋予抗性的宿主基因以及宿主基因感染所需的，通常在传统的基于 RNAi 的筛选中检测到。识别新靶点的潜力，包括非编码 RNA 元件，而使用现有的方法可能会错过这些靶点接近。