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，包括重要的人类病原体单纯疱疹病毒1 （HSV-1）和人畜共患病原体伪狂犬病毒（PRV），是极少数进化到利用高度特化的神经细胞生物学的病毒。在疾病的自然过程中，α疱疹病毒感染外周神经系统（PNS）的感觉和自主神经元。在再激活后，子代病毒颗粒在特定的亚细胞位点进行极化的细胞内运输和胞外分泌，从一个细胞扩散到另一个细胞，这是本提案的主题。在PNS神经元中，这种细胞内运输包括分选轴突，远距离运输到外周组织，导致疱疹或带状虫状病变复发，或中枢神经系统（CNS）。人类α -疱疹病毒，特别是1型单纯疱疹病毒，是病毒性脑炎的主要病因。在Aim 1中，我们将研究我们的初步数据表明在非神经元细胞中调节细胞内运输的病毒因子，在胞吐之前，使用现有的PRV试剂，并扩展到HSV-1。在Aim 2中，我们将研究高尔基后构成性分泌途径机制，该途径指导HSV-1颗粒运输和胞吐到非神经元细胞的特定亚细胞位点，并且我们假设它也介导神经元的极化运输和胞吐。利用创新的方法来急性干扰特定的细胞因子和直接成像病毒颗粒胞吐，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，比较非神经元细胞和原代PNS神经元。在Aim 3中，我们将重点关注介导轴突分选的微管运动机制，特别是在PNS神经元中。利用微流体样室神经元培养系统和活细胞成像，我们将确定不同的运动蛋白和微管相关蛋白在HSV-1颗粒轴突分选中的作用。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如何以及为何传播到神经系统和在神经系统内传播的理解，导致可药物靶点的鉴定和对病毒神经病理学的更好治疗的开发，并可能提供对细胞生物学，特别是神经元细胞生物学的基本见解。