Molecular and Cellular Mechanisms of Herpes Simplex Transport and Egress Pathway in Neurons

单纯疱疹神经元转运和出口途径的分子和细胞机制

• 批准号: 9243547
• 负责人: Ian B Hogue
• 金额: $ 15.4万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243547
• 关键词: Action Potentials; Address; Axon; Axonal Transport; Binding; Biological; Biological Assay; Biology; Capsid; Career Transition Award; Cell membrane; Cells; Cellular biology; Complex; Cryoelectron Microscopy; Cytoskeleton; Defect; Dendrites; Development; Disease; Dynein ATPase; Exocytosis; Fluorescence Microscopy; Fluorescent Probes; Funding; Goals; Golgi Apparatus; Herpes Simplex Infections; Herpes encephalitis; Herpesviridae; Herpesvirus 1; Human; In Vitro; Individual; Infection; Intercellular Junctions; Interneurons; Kinesin; Knowledge; Lead; Lesion; Life; Light; Mediating; Membrane; Methods; Microscopy; Microtubules; Molecular; Molecular Biology; Molecular Genetics; Motor; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurons; Neurovirology; Pathway interactions; Peripheral; Peripheral Nervous System; Phase; Positioning Attribute; Process; Proteins; Reagent; Recruitment Activity; Recycling; Reporting; Research; Resolution; Role; S-nitro-N-acetylpenicillamine; SNAP receptor; Secretory Cell; Secretory Vesicles; Sensory; Site; Structure; Structure-Activity Relationship; Surface; Synapses; Synaptic Vesicles; System; Tissues; United States National Institutes of Health; Vesicle; Viral; Virion; Virus; Virus Replication; base; career; education research; experience; in vivo; insight; instrumentation; latent infection; mad itch virus; neural circuit; neuronal transport; neuropathology; novel; particle; pathogen; predictive modeling; professor; programs; protein complex; rab GTP-Binding Proteins; spatiotemporal; structural biology; tomography; virus envelope

Project Summary/Abstract The alpha herpesviruses, which includes important human pathogen herpes simplex virus 1 (HSV-1), are among the very few viruses that have evolved neuron-specific viral mechanisms to exploit highly-specialized neuronal cell biology. During the natural course of disease, alpha herpesviruses infect sensory or autonomic neurons, and establish a life-long latent infection in the peripheral nervous system. Upon reactivation, the virus can return to peripheral tissues, causing herpetic or zosteriform lesions, or it can spread to the central nervous system, causing severe herpes encephalitis. This proposal focuses on three major steps in the HSV-1 replication cycle, where virus particles must interact with specialized neuronal systems: 1. long-distance post- entry axonal transport; 2. long-distance post-replication transport of progeny particles towards axonal sites of egress; 3. polarized viral egress via exocytosis from axons, dendrites, or cell bodies. In Aim 1, we will identify sites of HSV-1 particle exocytosis in neurons, and determine the molecular/cellular mechanisms of HSV-1 egress from axons, dendrites, or cell bodies. In Aim 2, we will investigate the structural biology of HSV-1 axonal transport by cryo electron microscopy (cryoEM). My prior education and research experience has focused on studying viruses from a molecular and cell biological perspective, in particular, studying the interactions of enveloped viruses with host membrane systems. In the course of my graduate and postdoctoral research, I have developed expertise in using specialized microscopy methods to spatiotemporally dissect particular steps in the virus replication cycle. In the current postdoctoral phase of my career, I have developed a novel live-cell fluorescence microscopy assay of virus egress based on a specialized fluorescence microscopy method and a novel pH-sensitive fluorescent probe. In addition, I am currently developing new cryoEM tomography methods to investigate axonal transport of virus particles. This proposal seeks to extend these methods to accomplish the proposed aims. My scientific background makes me well suited to carry out the proposed research. This K22 Career Transition Award will help me to achieve my scientific and career goals, which include transitioning to an independent position as an assistant professor, and establishing an independent NIH-funded research program focused on neurovirology.

项目摘要/摘要 α疱疹病毒，包括重要的人类病原体疱疹病毒1（HSV-1），是 在几乎没有发展神经元特异性病毒机制以利用高度特异性的病毒中 神经元细胞生物学。在自然疾病过程中，α疱疹病毒感染感觉或自主神经 神经元，并在周围神经系统中建立终生的潜在感染。重新激活后，病毒 可以返回到周围组织，导致疱疹或暗膜病变，或者可以扩散到中枢神经 系统，导致严重的疱疹脑炎。该提案重点介绍了HSV-1的三个主要步骤 复制周期，病毒颗粒必须与专门的神经元系统相互作用：1。 进入轴突运输； 2。后代颗粒的恢复后长距离转运向轴突部位 出口； 3。通过轴突，树突或细胞体的胞吐作用通过偏振病毒出口。在AIM 1中，我们将确定 神经元中HSV-1颗粒胞吐作用的位点，并确定HSV-1的分子/细胞机制 轴突，树突或细胞体的出口。在AIM 2中，我们将研究HSV-1的结构生物学 冷冻电子显微镜（冷冻）的轴突运输。我先前的教育和研究经验 尤其是从分子和细胞生物学角度研究病毒的重点，特别是研究 包裹病毒与宿主膜系统的相互作用。在我的毕业生和博士后 研究，我已经开发了使用专门显微镜方法的专业知识来时空剖析 病毒复制周期的特定步骤。在我职业生涯的当前博士后阶段，我已经开发了 一种基于专业荧光的病毒出口的新型活细胞荧光显微镜测定 显微镜方法和一种新型的pH敏感荧光探针。此外，我目前正在开发新的 冷冻断层扫描方法研究病毒颗粒的轴突运输。该建议旨在扩展 这些方法是完成所提出的目标。我的科学背景使我非常适合执行 拟议的研究。这个K22职业过渡奖将帮助我实现我的科学和职业 目标包括过渡到助理教授的独立职位，并建立一个 独立的NIH资助研究计划的重点是神经病毒学。