Intersection of HSV latency and reactivation with the neuronal apoptotic pathway

HSV 潜伏期和再激活与神经元凋亡途径的交叉点

基本信息

批准号：
8316708
负责人：
Anna Ruth Cliffe
金额：
$ 5.94万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8316708
关键词：
Accounting Animal Model Apoptosis Apoptotic Arts Cell Death Cell Nucleus Cells Characteristics DNA Damage Data Disease Drug Delivery Systems Event Family Functional RNA Gene Expression Genes Goals Immune response Immunocompromised Host In Vitro Individual Induction of Apoptosis Infection Injection of therapeutic agent Introns JUN gene Knowledge Lytic Microinjections Mitochondria Modeling Molecular Morbidity - disease rate Mus Nerve Growth Factors Neuroglia Neurons Pathway interactions Pattern Plasmids Protein Biosynthesis Proteins RNA Regulation Role Signal Transduction Simplexvirus Small RNA Staging Stimulus Stress Techniques Testing Transcript Viral Viral Genes Viral Proteins Virus Virus Latency caspase-3 caspase-9 cell type cytochrome c deprivation gene function granzyme B in vitro Model in vivo knock-down latency associated transcript latent infection lytic gene expression mortality neuron apoptosis neuron loss new therapeutic target novel prevent reactivation from latency research study response transcription factor

项目摘要

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) persists for the lifetime of the host in the form of a latent infection of neurons. Importantly, periodic reactivation of the vius results in significant morbidity and mortality, particularly in the immunocompromised host. However, the unique characteristics of neurons and molecular events that allow viral persistence and its reactivation are not understood. So far, obstacles to understanding the interaction of the virus with neurons have been i) the limited use of an in vitro model of latency and ii) challenges in gene manipulation techniques in primary neurons. Infection of sympathetic neurons has been found to recapitulate HSV latency in vivo. Therefore, in this project I will use sympathetic neurons and state-of-art techniques to examine viral gene function, and the mechanism of viral reactivation at the molecular and cellular level in neurons. The latency-associated transcript (LAT) encodes a family of non-coding RNAs and is the only viral gene product expressed to high levels during latency. In Aim 1, I will test the hypothesis that LAT expression inhibits apoptosis and promotes survival of infected neurons, thus allowing for long-term viral persistence and enhanced reactivation. LAT expressing plasmids will be introduced into neurons by microinjection. The ability of the LAT to protect neurons against different triggers of apoptosis and the mechanism by which the LAT exerts protection will also be determined. In Aim 2, I will focus on examining the signaling events within neurons that trigger HSV reactivation. Viral reactivation is triggered when sympathetic neurons are deprived of nerve growth factor (NGF). Since NGF deprivation activates apoptosis in neurons, I will identify the key event in the apoptotic pathway after NGF deprivation that activates the expression of HSV lytic genes to allow viral reactivation. An understanding of how HSV latency is maintained at the cellular level and knowledge of key events within neurons that trigger its reactivation are critica to identify potential targets for novel therapeutics that prevent HSV reactivation from neurons. PUBLIC HEALTH RELEVANCE: Herpes Simplex Virus is able to hide for the lifetime of an individual in the form of a latent infection of neurons. The experiments proposed in this project will help uncover how the virus is able to persist and what triggers the virus to reactivate from neurons and cause disease. The long-term goals are to identify new targets for drugs that would prevent the reactivation of Herpes Simplex Virus.

描述（由申请人提供）：单纯疱疹病毒（HSV）以神经元潜伏感染的形式在宿主的一生中持续存在。重要的是，vius的周期性重新激活会导致显着的发病率和死亡率，特别是在免疫功能低下的宿主中。然而，导致病毒持续存在及其重新激活的神经元和分子事件的独特特征尚不清楚。到目前为止，理解病毒与神经元相互作用的障碍是 i) 体外潜伏模型的有限使用和 ii) 原代神经元基因操作技术的挑战。已发现交感神经元的感染可再现体内 HSV 潜伏期。因此，在这个项目中，我将使用交感神经元和最先进的技术来检查病毒基因功能，以及在神经元的分子和细胞水平上病毒再激活的机制。潜伏期相关转录本 (LAT) 编码非编码 RNA 家族，是唯一在潜伏期高水平表达的病毒基因产物。在目标 1 中，我将检验 LAT 表达抑制细胞凋亡并促进受感染神经元存活的假设，从而实现病毒的长期持续存在并增强重新激活。 LAT表达质粒将通过显微注射引入神经元中。 LAT 保护神经元免受不同凋亡触发因素的能力以及 LAT 发挥保护作用的机制也将被确定。在目标 2 中，我将重点研究神经元内触发 HSV 重新激活的信号事件。当交感神经元缺乏神经生长因子（NGF）时，就会触发病毒重新激活。由于 NGF 剥夺会激活神经元细胞凋亡，因此我将确定 NGF 剥夺后细胞凋亡途径中激活 HSV 裂解基因表达以允许病毒重新激活的关键事件。了解 HSV 潜伏期如何在细胞水平上维持，以及了解神经元内触发其重新激活的关键事件，对于确定防止神经元 HSV 重新激活的新型疗法的潜在靶标至关重要。公共卫生相关性：单纯疱疹病毒能够以神经元潜伏感染的形式在个体一生中隐藏。该项目提出的实验将有助于揭示病毒如何能够持续存在，以及是什么触发病毒从神经元重新激活并导致疾病。长期目标是确定新的药物靶标，以防止单纯疱疹病毒重新激活。