Mechanisms of alphavirus infectivity and adaptation - Resubmission - 1

甲病毒感染性和适应机制 - 重新提交 - 1

• 批准号: 10444392
• 负责人: Kenneth Stapleford
• 金额: $ 53.24万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10444392
• 关键词: Aedes; Alphavirus; Antiviral Agents; Antiviral Therapy; Arbovirus Infections; Arboviruses; Arthropod Vectors; Biochemical Genetics; Biology; Cells; Chikungunya virus; Cholesterol; Coupled; Culicidae; Disease; Environmental Risk Factor; Epidemic; Event; Evolution; Genetic; Glycoproteins; Goals; Human; In Vitro; Infection; Insect Vectors; Insecta; Integration Host Factors; Knowledge; Lead; Life Cycle Stages; Lipids; Mammalian Cell; Mammals; Molecular; Mus; Mutation; Pathogenesis; Play; Process; Proteins; Public Health; Role; Saliva; Structural Protein; Technology; Temperature; Testing; Vaccine Therapy; Variant; Viral; Viral Pathogenesis; Viral Structural Proteins; Virion; Virulence; Virus; Work; antiviral drug development; base; deep sequencing; driving force; emerging human pathogen; in vivo; novel; protein function; targeted treatment; thermostability; transmission process; vaccine development; vector; viral transmission

PROJECT SUMMARY Arboviruses are transmitted from insect vectors to humans, yet the molecular mechanisms that govern this critical step in the viral life cycle are not completely understood. Chikungunya virus (CHIKV) has undergone several natural adaptation events, acquiring mutations in the viral glycoproteins that have increased its infectivity and transmission, yet how the glycoproteins promote infectivity, spread, and disease in vivo is unclear. We harnessed the power of in vivo viral evolution and identified a new evolutionary intermediate in β- strand c of the CHIKV E1 glycoprotein that led to increased transmission in mosquitoes and increased replication and pathogenesis in mice, indicating that these glycoprotein regions are crucial for promoting viral infectivity and disease. Preliminary studies from our lab show that E1 β-strand c plays essential roles in virion thermostability, fusion, and cholesterol-dependent entry. Moreover, using viral evolution we have identified host-specific genetic interactions between CHIKV E1 and E2. Based on these preliminary studies, we hypothesize that CHIKV E1 and E2 drive distinct mechanisms of entry into insects and mammals and that temperature and lipid composition in mosquitoes drives viral adaptation. Our overall goal is to understand the fundamental mechanisms by which the CHIKV structural proteins regulate alphavirus infectivity and dissemination in insects and mammals, and to elucidate what drives the adaptation of the glycoprotein in insects. Here, we use complementary biochemical, genetic, in vitro and in vivo approaches coupled with deep sequencing technologies to establish how the CHIKV structural proteins function to promote entry and CHIKV pathogenesis and transmission in vivo (Aim 1), and to establish how temperature and host lipids impact CHIKV-vector interactions in mosquitoes (Aim 2). Understanding the discrete mechanisms of how the viral structural proteins regulate infectivity in insects and mammals, and how host lipids impact adaptation is essential to understanding how arboviruses are transmitted, spread, and cause disease.

项目概要 虫媒病毒从昆虫媒介传播到人类，但控制这种病毒的分子机制 基孔肯雅病毒（CHIKV）病毒生命周期中的关键步骤尚未完全了解。 一些自然适应事件，病毒糖蛋白发生突变，从而增加了其 传染性和传播，但糖蛋白如何在体内促进传染性、传播和疾病尚不清楚 我们利用体内病毒进化的力量，在 β- 中发现了一种新的进化中间体。 CHIKV E1 糖蛋白的 c 链导致蚊子中的传播增加并增加 小鼠体内的复制和发病机制，表明这些糖蛋白区域对于促进病毒传播至关重要 我们实验室的初步研究表明，E1 β-链 c 在病毒颗粒中发挥着重要作用。 此外，利用病毒进化，我们已经确定了热稳定性、融合和胆固醇依赖性进入。 基于这些初步研究，我们发现了 CHIKV E1 和 E2 之间的宿主特异性遗传相互作用。 CHIKV E1 和 E2 驱动不同的进入昆虫和哺乳动物的机制 我们的总体目标是了解蚊子的温度和脂质成分驱动病毒的适应。 CHIKV 结构蛋白调节甲病毒感染性的基本机制 在昆虫和哺乳动物中传播，并阐明驱动糖蛋白适应的因素 在这里，我们使用互补的生化、遗传、体外和体内方法以及深层方法。 测序技术以确定 CHIKV 结构蛋白如何发挥作用以促进进入和 CHIKV 体内发病机制和传播（目标 1），并确定温度和宿主脂质如何影响 蚊子中的 CHIKV 与载体相互作用（目标 2）。 结构蛋白调节昆虫和哺乳动物的感染性，以及宿主脂质如何影响适应性 对于了解虫媒病毒如何传播、传播和引起疾病至关重要。