Identification of host factors required by the tick-borne Powassan virus

蜱传波瓦桑病毒所需宿主因子的鉴定

• 批准号: 10154884
• 负责人: Charles M Rice
• 金额: $ 25.43万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-11-23 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10154884
• 关键词: Antiviral Agents; Arboviruses; Biology; Bite; CRISPR screen; Candidate Disease Gene; Cell Line; Cells; Centers for Disease Control and Prevention (U.S.); Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Confusion; Development; Disease; Drug Targeting; Drug resistance; Ecosystem; Encephalitis; FDA approved; Fever; Flaviviridae; Flavivirus; Flavivirus Infections; Future; Genes; Genetic; Genomic Library; Geography; Goals; Headache; Health; Hemiplegia; Hemorrhage; Human; Human Genome; Impairment; Infection; Infection Control; Integration Host Factors; Intervention; Invertebrates; Ixodes; Knock-out; Measures; Memory; Memory Loss; Morbidity - disease rate; Motor Skills Disorders; Muscular Atrophy; Mutation; Natural Immunity; Neurologic Symptoms; Patients; Polymerase; Powassan virus; Proteins; Readiness; Reporting; Ribavirin; Role; Seizures; Seroprevalences; Structure; Supportive care; Symptoms; Technology; Therapeutic; Therapeutic Intervention; Ticks; Viral Hemorrhagic Fevers; Virus; Virus Diseases; Virus Replication; Vomiting; Work; ZIKA; acaricide; adaptive immunity; arthropod-borne; clinical phenotype; design; drug development; genome-wide; infection risk; insight; invertebrate host; member; mortality; mosquito-borne; prevent; prophylactic; tick-borne; tick-borne flavivirus; tissue tropism; vector tick; warm temperature

PROJECT SUMMARY Flaviviruses cause significant morbidity and mortality worldwide. The sudden emergence and spread of viruses such as Zika illustrate our vulnerability and emphasize the need for preparedness against related emerging viruses. In addition to mosquito-borne flaviviruses, members of tick-borne flaviviruses pose an increasing global threat for which we lack effective antivirals. Powassan virus (POWV) is an emerging virus transmitted by the bite of infected Ixodes ticks. The seroprevalence of POWV in humans is mostly unknown as infections may often be asymptomatic. However, symptomatic POWV infections may progress in to severe and sometimes fatal encephalic disease with mortality rates of ~10% and surviving patients often suffer from debilitating long-term neurologic symptoms. The severe clinical consequences of infections with POWV and other tick-borne flaviviruses and the lack of specific treatments highlight an urgent need for a better understanding of these viruses in their tick and mammalian hosts so that preventative and therapeutic treatments can be developed. The goal of this project is to identify host factors required by POWV that may also impact a wide range of existing and emerging flaviviruses. These host proteins may represent suitable targets for antiviral interventions, and we hypothesize that impaired virus replication due to their inhibition may allow innate and adaptive immunity to control the infection. Further, we propose to compare a panel of flaviviruses, including POWV, and their requirements for the identified host factors in both vertebrate and invertebrate cells. We believe these approaches will allow us to prioritize candidate genes for future drug development studies. The proposed studies will utilize CRISPR/Cas9 screening technology to knock out every gene in the human genome with the goal of identifying host factors that facilitate POWV infection. We will use a similar approach in a targeted, arrayed format to evaluate the identified host factors in various human and tick cell lines during infection with diverse flaviviruses. We expect that this work will broaden our understanding of tick- and mosquito-borne flaviviruses, specifically in terms of clinical phenotype (encephalitic vs hemorrhagic disease) and the shared need for host factors. In addition, it will help to define which genes govern host species and tissue tropism. Together, this work will provide a wealth of new insights into many aspects of flavivirus biology.

项目概要 黄病毒在世界范围内引起显着的发病率和死亡率。病毒的突然出现和传播 诸如寨卡病毒之类的事件说明了我们的脆弱性，并强调了针对相关新兴疾病做好准备的必要性 病毒。除了蚊媒黄病毒外，蜱媒黄病毒的成员在全球范围内也日益增多。 我们缺乏有效的抗病毒药物来应对这种威胁。 波瓦桑病毒 (POWV) 是一种新出现的病毒，通过受感染的硬蜱叮咬传播。这 人类中 POWV 的血清流行率大多未知，因为感染通常可能无症状。然而， 有症状的 POWV 感染可能会发展为严重且有时致命的脑部疾病，导致死亡 死亡率约为 10%，幸存的患者经常患有使人衰弱的长期神经系统症状。严重者 POWV 和其他蜱传黄病毒感染的临床后果以及缺乏特异性 治疗凸显了迫切需要更好地了解蜱虫和哺乳动物宿主中的这些病毒 以便开发预防和治疗方法。 该项目的目标是确定 POWV 所需的宿主因素，这些因素也可能影响广泛的 现有和新出现的黄病毒。这些宿主蛋白可能代表抗病毒的合适靶点 干预措施，我们假设由于其抑制而导致病毒复制受损可能会导致先天和 适应性免疫来控制感染。此外，我们建议比较一组黄病毒，包括 POWV，以及它们对脊椎动物和无脊椎动物细胞中已确定的宿主因子的要求。我们相信 这些方法将使我们能够优先考虑未来药物开发研究的候选基因。 拟议的研究将利用 CRISPR/Cas9 筛选技术来敲除细胞中的每个基因。 人类基因组，目标是识别促进 POWV 感染的宿主因素。我们将使用类似的 采用有针对性的阵列形式的方法来评估各种人类和蜱细胞系中已识别的宿主因素 在感染多种黄病毒期间。 我们期望这项工作将扩大我们对蜱和蚊媒黄病毒的了解， 特别是在临床表型（脑炎与出血性疾病）和对宿主的共同需求方面 因素。此外，它将有助于确定哪些基因控制宿主物种和组织向性。共同完成这项工作 将为黄病毒生物学的许多方面提供丰富的新见解。