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：
8628197
负责人：
Anna Ruth Cliffe
金额：
$ 6.31万
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/8628197
关键词：
Accounting Animal Model Apoptosis Apoptotic Arts Cell Death Cell Nucleus Cells Characteristics DNA Damage Data Disease Drug Targeting 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 MicroRNAs 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.

描述（由申请人提供）：单纯疱疹病毒（HSV）在宿主的一生中一直以神经元的潜在感染形式存在。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。但是，尚不清楚允许病毒持久性及其重新激活的神经元和分子事件的独特特征。到目前为止，理解病毒与神经元相互作用的障碍是i）有限使用潜伏期的体外模型和II）原发性神经元中基因操纵技术的挑战。已发现感染交感神经元可以概括体内的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检查病毒基因功能，以及在神经元分子和细胞水平上病毒重新激活的机制。延迟相关的转录本（LAT）编码非编码RNA的家族，是唯一在潜伏期中表达高水平的病毒基因产物。在AIM 1中，我将检验以下假设：LAT表达抑制凋亡并促进感染神经元的存活，从而允许长期的病毒持久性和增强的重新激活。 LAT表达质粒将通过显微注射引入神经元。 LAT保护神经元免受不同凋亡触发因素的能力以及LAT施加保护的机制也将得到确定。在AIM 2中，我将专注于检查触发HSV重新激活的神经元内的信号事件。当交感神经元被剥夺神经生长因子（NGF）时，会触发病毒重新激活。由于NGF剥夺激活了神经元中的凋亡，因此我将确定NGF剥夺后凋亡途径中的关键事件，该事件激活了HSV裂解基因的表达以允许病毒重新激活。对HSV潜伏期如何保持在细胞水平和触发其重新激活的关键事件的知识的理解是批评的，批评了防止神经元的HSV重新激活的新型治疗剂的潜在靶标。