LDLRAD3 Receptor Interaction with Venezuelan Equine Encephalitis Virus

LDLRAD3 受体与委内瑞拉马脑炎病毒的相互作用

• 批准号: 10435558
• 负责人: Michael S Diamond
• 金额: $ 76.27万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10435558
• 关键词: Acute; Acute Disease; Aerosols; Africa; Alphavirus; Alphavirus Infections; Americas; Animals; Antibodies; Antiviral Agents; Binding; Binding Sites; Biochemical; Biological Assay; Biology; Biophysics; CRISPR/Cas technology; Cell membrane; Cell surface; Cells; Chikungunya virus; Chronic; Complement; Confocal Microscopy; Cryoelectron Microscopy; Culicidae; Data; Development; Diamond; Disease; Disease Outbreaks; Disease Outcome; Dissection; Ectopic Expression; Epidemic; Epitopes; Equus caballus; Family; Fc Receptor; Fever; Future; Genes; Genetic; Genetic study; Glycoproteins; Goals; Human; Immunoelectron Microscopy; Infection; Integration Host Factors; Knockout Mice; Label; Laboratories; Libraries; Mediating; Medical; Membrane; Molecular Conformation; Morphogenesis; Mus; Mutagenesis; N-terminal; Nature; Neurons; Oceania; Pathogenesis; Pathogenicity; Pathway interactions; Penetration; Polyarthritides; Prevention; Proteins; RNA Viruses; Reagent; Recombinants; Resolution; Role; Route; Structure; Structure-Activity Relationship; Testing; Therapeutic; Tissues; Togaviridae; Translating; Tropism; Universities; Vaccines; Venezuelan Equine Encephalitis Virus; Vertebrates; Vesicular stomatitis Indiana virus; Video Microscopy; Viral; Viral Pathogenesis; Virion; Virus; Virus Diseases; Washington; Western Equine Encephalitis Virus; X-Ray Crystallography; acute infection; base; biological development; cell type; defined contribution; emerging pathogen; env Gene Products; epizootic; experimental study; genetic analysis; genome-wide; in vivo; innovation; insight; medical countermeasure; member; mouse model; mutant; novel; novel strategies; pathogenic virus; prevent; receptor; receptor internalization; scavenger receptor; small molecule; tissue tropism; virology

Project Summary Alphaviruses are mosquito-transmitted, positive-strand enveloped RNA viruses of the Togaviridae family that cause global disease in humans. At present, no antiviral agents or licensed vaccines exist for the treatment or prevention of any alphavirus infections. We recently used a genome-wide CRISPR/Cas9-based screen to identify the cell surface molecule LDLRAD3 as a novel, highly conserved entry receptor for Venezuelan equine encephalitis virus (VEEV), an emerging pathogen capable of causing fatal neuroinvasive disease in humans and other vertebrate animals. Gene editing of mouse or human LDLRAD3 resulted in reduced VEEV infection of neuronal cells, and reciprocally, ectopic expression of LDLRAD3 resulted in increased infection. LDLRAD3 bound directly to VEEV virions and enhanced virus attachment and internalization into cells. Genetic studies indicated that domain 1 (D1) of LDLRAD3 is necessary and sufficient to support VEEV infection. We hypothesize that engagement of LDLRAD3 by VEEV will explain how infection, tissue targeting, and disease pathogenesis occurs. The primary goals of this collaborative, interactive project between the Diamond, Fremont, and Whelan laboratories are to define the precise mechanism(s) by which LDLRAD3 facilitates alphavirus entry into cells, to gain high-resolution structural insight as to how LDLRAD3 engages the spike proteins on the virion, and to determine the cell-type specific role of LDLRAD3 in VEEV pathogenesis in vivo. The experiments in this proposal will define fundamental aspects of VEEV biology that enhance our understanding of infection and cell tropism. This information may facilitate the development of small molecules or biologicals that disrupt LDLRAD3 interaction with VEEV spike proteins, which could form the basis of future therapeutics that ameliorate disease of this emerging and highly pathogenic alphavirus.

项目概要 甲病毒是披膜病毒科的由蚊子传播的、有正链包膜的 RNA 病毒， 导致人类全球性疾病。目前，尚无用于治疗或治疗的抗病毒药物或许可的疫苗。 预防任何甲病毒感染。我们最近使用基于 CRISPR/Cas9 的全基因组筛选来识别 细胞表面分子 LDLRAD3 作为委内瑞拉马的一种新型、高度保守的进入受体 脑炎病毒（VEEV）是一种新出现的病原体，能够引起人类致命的神经侵袭性疾病 其他脊椎动物。小鼠或人类 LDLRAD3 的基因编辑导致 VEEV 感染减少 LDLRAD3 的异位表达反过来导致感染增加。低密度脂蛋白RAD3 直接与 VEEV 病毒粒子结合，增强病毒附着和内化到细胞中。遗传学研究 表明 LDLRAD3 的结构域 1 (D1) 对于支持 VEEV 感染是必要且充分的。我们假设 VEEV 与 LDLRAD3 的结合将解释感染、组织靶向和疾病发病机制 发生。 Diamond、Fremont 和 Whelan 之间合作、互动项目的主要目标 实验室将确定 LDLRAD3 促进甲病毒进入细胞的精确机制，以 获得关于 LDLRAD3 如何与病毒体上的刺突蛋白结合的高分辨率结构洞察，并 确定 LDLRAD3 在体内 VEEV 发病机制中的细胞类型特异性作用。本提案中的实验 将定义 VEEV 生物学的基本方面，以增强我们对感染和细胞向性的理解。 这些信息可能会促进破坏 LDLRAD3 的小分子或生物制品的开发 与 VEEV 刺突蛋白相互作用，这可能构成未来改善疾病疗法的基础 这种新兴的高致病性甲病毒。