Molecular and cellular mechanisms of HSV-1 assembly and egress

HSV-1 组装和流出的分子和细胞机制

• 批准号: 10366928
• 负责人: Ian B Hogue
• 金额: $ 35.85万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10366928
• 关键词: Acute; Address; Agricultural Workers; Axon; Biological Models; Biological Process; Biology; Cell membrane; Cells; Cellular biology; Characteristics; China; Data; Development; Disease; Distal; Encephalitis; Exhibits; Exocytosis; Family suidae; Fluorescence Microscopy; Focal Adhesions; Genes; Goals; Golgi Apparatus; Herpesviridae; Herpesvirus 1; Human; Image; Integration Host Factors; Intracellular Membranes; Intracellular Transport; Kinesin; Knowledge; Lead; Lesion; Light; Mediating; Methods; Microfluidics; Microtubule-Associated Protein 2; Microtubule-Associated Proteins; Microtubules; Modeling; Molecular; Molecular Biology; Motor; Mutation; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurons; Pathway interactions; Peripheral; Peripheral Nervous System; Pharmaceutical Preparations; Phenotype; Population; Process; Reagent; Recurrence; Reporting; Research; Role; Secretory Vesicles; Sensory; Site; Sorting - Cell Movement; System; Tissues; Vaccines; Vesicle; Viral; Viral Encephalitis; Viral Matrix Proteins; Viral Proteins; Virion; Virus; Zoonoses; base; druggable target; gE protein; human pathogen; innovation; insight; knock-down; live cell imaging; mad itch virus; mutant; neuroinflammation; neuropathology; particle; pathogen; recruit; trafficking

Alpha herpesviruses, including important human pathogen Herpes Simplex Virus 1 (HSV-1) and zoonotic pathogen Pseudorabies Virus (PRV), are among the very few viruses that have evolved to exploit highly-specialized neuronal cell biology. During the natural course of disease, alpha herpesviruses infect sensory and autonomic neurons of the Peripheral Nervous System (PNS). Upon reactivation, progeny virus particles undergo polarized intracellular trafficking and exocytosis at particular sub-cellular sites to spread from cell to cell, which is the subject of this proposal. In PNS neurons, this intracellular trafficking can include sorting into axons for long-distance transport into peripheral tissues, leading to recurrence of herpetic or zosteriform lesions, or to the Central Nervous System (CNS). The human alpha herpesviruses, and HSV-1 in particular, are leading causes of viral encephalitis. In Aim 1, we will investigate the viral factors that our preliminary data suggest modulate intracellular transport in non-neuronal cells, immediately prior to exocytosis, using existing reagents in PRV, and extending to HSV-1. In Aim 2, we will investigate the post-Golgi constitutive secretory pathway mechanisms that direct HSV-1 particle trafficking and exocytosis to particular sub-cellular sites in non- neuronal cells, and which we hypothesize also mediate polarized trafficking and exocytosis in neurons. Using innovative methods to acutely perturb particular cellular factors and directly image virus particle exocytosis, we will determine the role of secretory pathway mechanisms in intracellular trafficking and egress of virus particles, comparing non-neuronal cells to primary PNS neurons. In Aim 3, we will focus on the microtubule motor-based mechanisms that mediate axonal sorting, specifically in PNS neurons. Using a microfluidics-like chambered neuronal culture system and live-cell imaging, we will determine the roles of different kinesin motors and microtubule-associated proteins in axonal sorting of HSV-1 particles. Elucidating the basic cell biological processes that our viruses use in both neurons and non-neuronal cells will increase our understanding of how and why herpesviruses spread to and within the nervous system, lead to the identification of druggable targets and development of better therapies for viral neuropathology, and may provide fundamental insights into cell biology, particularly of the cell biology of neurons.

α疱疹病毒，包括重要的人病原体疱疹病毒1（HSV-1）和人畜共患病原体假病毒（PRV），是少数进化以利用高度特异性神经元细胞生物学的病毒之一。在自然疾病过程中，α疱疹病毒感染周围神经系统（PNS）的感觉和自主神经元。重新激活后，后代病毒颗粒会在特定的亚细胞部位进行极化的细胞内运输和胞吐作用，从而从细胞到细胞扩散，这是该建议的主题。在PNS神经元中，这种细胞内运输可以包括将轴突分类为长距离运输到外周组织，从而导致疱疹或灵活的病变的复发，或中枢神经系统（CNS）。人α疱疹病毒，尤其是HSV-1是病毒脑炎的主要原因。在AIM 1中，我们将研究我们的初步数据表明在胞吐作用之前，使用PRV中的现有试剂并延伸至HSV-1的病毒因素表明我们的初步数据表明在非神经元细胞中调节细胞内转运。在AIM 2中，我们将研究Golgi后的分泌后分泌途径机制，这些机制将HSV-1颗粒运输和胞吐作用引导到非神经元细胞中的特定亚细胞部位，并且我们假设该神经元还介导了神经元中的极化运输和胞外增生。使用创新的方法来急性扰动特定的细胞因子并直接成像病毒颗粒胞吐作用，我们将确定分泌途径机制在细胞内运输和病毒颗粒出口中的作用，将非神经元细胞与原发性PNS神经元进行比较。在AIM 3中，我们将专注于介导轴突分选的微管运动机制，特别是在PNS神经元中。使用类似于微流体的腔室神经元培养系统和活细胞成像，我们将确定不同驱动蛋白电机和微管相关蛋白在HSV-1颗粒的轴突分类中的作用。阐明我们的病毒在神经元和非神经元细胞中使用的基本细胞生物学过程将增加我们对疱疹病毒的理解以及为什么扩散到神经系统内部和内部的原因，导致可识别可药物的鉴定并开发出更好的病毒神经病理学疗法，并可能为细胞生物学提供基本的洞察力，尤其是Neurons of Neurons。