HSV latency and reactivation and the novel neuronal regulation of VP16 in vivo.

HSV 潜伏期和再激活以及体内 VP16 的新神经元调节。

基本信息

批准号：
8868009
负责人：
Nancy M. Sawtell
金额：
$ 50.39万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
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：
2012-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8868009
关键词：
Acute Affect Afferent Neurons Antiviral Agents Binding Bioinformatics Blindness Cardiovascular Diseases Cells Chronic Data Development Diabetes Mellitus Disease Early Gene Transcriptions Elements Encephalitis Enzymes Event Frequencies Future Gene Activation Gene Expression Gene Silencing Genes Genetic Genome Goals HIV Infections Herpes Simplex Virus Protein Vmw65 Herpesvirus 1 Human Human Herpesvirus 2 Immediate-Early Genes In Vitro Infection Inflammation Integration Host Factors Intervention Knowledge Latent Virus Lead Lytic Phase Mediating Mutation Nervous system structure Neuraxis Neurodegenerative Disorders Neurons Nutrient Outcome Pattern Phosphorylation Phosphorylation Site Phosphotransferases Population Post-Translational Protein Processing Prevention strategy Process Proteins Recurrence Regulation Regulatory Element Research Role Signal Transduction Simplexvirus Site Stress Testing Therapeutic Intervention Transactivation VP 16 Vaccines Viral Viral Genome Viral Proteins Virion Virulence Virus Virus Diseases Virus Latency Work biological adaptation to stress design host cell factor C1 human CREB1 protein improved in vivo latent infection lytic replication mouse model novel prevent promoter protein expression reactivation from latency recombinant virus response sensor transcription factor treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Most of the human population world-wide has been infected by herpes simplex viruses. Following the initial lytic infection, HSVs establish permanent latent infections within sensory neurons. Reactivation of latent virus not only results in viral disease (new infections, blindness, and encephalitis) but also contributes to HIV infection, diabetes, cardiovascular and neurodegenerative diseases. No effective vaccine is available and no therapy eliminates latency or prevents reactivation. The long-term goal of this project is to find interventions for recurrent HSV episodes by defining mechanisms that control establishment and reactivation of HSV-1 latency. The gene expression cascade during HSV-1 lytic infection begins with activation of immediate-early (IE) gene transcription by the virion protein VP16 with host factors Oct-1 and HCF-1. In contrast, the initial events in the reactivation from latency are still poorly defined. The central hypothesis of this proposal is that regulation o both VP16 expression and activity underlie the establishment of latency and reactivation from latency. These two levels of control involve multiple positive and negative inputs to allow or inhibit viral replication in the sensory neuron in vivo. Aim 1. This project will determine the mechanism of de novo VP16 gene activation and silencing in sensory neurons in vivo. The working hypothesis is that the VP16 gene in the HSV-1 genome can be regulated by action of neuron-specific and stress-responsive promoter elements and corresponding transcription factors either to allow or inhibit lytic replication upon initial infection or exit from latency. Uing recombinant viruses in a mouse model of infection and latency, we found that expression of VP16 is both necessary and sufficient to trigger the exit from latency and we identified a neuron-specific promoter for the VP16 gene. We will test whether this promoter controls the entry into lytic phase infection during acute infection and during reactivation. We will determine whether the predicted transcription factors bind to the various elements of this promoter to positively or negatively regulate VP16 gene expression, whether singly, in combination, or in competition. Aim 2. This project will define the VP16 coactivator interactions essential for VP16-dependent exit from latency and identify mechanisms regulating these interactions in vivo. Our data strongly suggest that the exit from latency by HSV is regulated by CK2 mediated phosphorylation and that this phosphorylation may be also competitively regulated by O-GlcNAcylation (a PTM that regulates signaling in response to nutrients and stress). Our goal is to elucidate the functions of each of these PTMs in viral latency and to define the roles of their crosstalk in regulating immediate early gene expression of viral proteins through VP16 transactivation. The outcomes of this work will identify transcription factors or protein modifying enzymes that could be targets for future development of therapeutic interventions for HSV reactivation.

描述（由申请人提供）：全世界大多数人口都被单纯疱疹病毒感染。在初始裂解感染之后，HSV在感觉神经元内建立永久性潜在感染。潜在病毒的重新激活不仅导致病毒疾病（新感染，失明和脑炎），还导致HIV感染，糖尿病，心血管和神经退行性疾病。没有有效的疫苗可用，也没有治疗可以消除潜伏期或阻止重新激活。该项目的长期目标是通过定义控制HSV-1潜伏期的建立和重新激活的机制来找到复发性HSV发作的干预措施。 HSV-1裂解感染期间的基因表达级联反应始于病毒粒子蛋白VP16的立即（IE）基因转录，具有宿主因子Oct-1和HCF-1。相反，重新激活中的初始事件从潜伏期起的定义仍然很差。该提议的中心假设是，VP16表达和活性的调节是建立潜伏期和延迟重新激活的基础。这两个级别的控制涉及多个阳性和负输入，以允许或抑制体内感觉神经元中的病毒复制。 AIM 1。该项目将确定体内感觉神经元中的从头VP16基因激活和沉默的机制。工作假设是，HSV-1基因组中的VP16基因可以通过神经元特异性和应激响应性启动子元素和相应的转录因子的作用来调节，以允许或抑制初始感染后的裂解复制或退出延迟。在小鼠感染和潜伏期的小鼠模型中，UING重组病毒，我们发现VP16的表达既需要又足以触发潜伏期的退出，我们确定了VP16基因的神经元特异性启动子。我们将测试该启动子在急性感染期间和重新激活过程中是否控制进入裂解相感染。我们将确定预测的转录因子是否与该启动子的各个元素结合，以积极或负调节VP16基因表达，无论是单独的，组合还是在竞争中。 AIM 2。该项目将定义VP16共激活因子相互作用对于依赖延迟的VP16依赖性退出所必需的，并确定在体内调节这些相互作用的机制。我们的数据强烈表明，HSV从潜伏期中退出受CK2介导的磷酸化调节，并且这种磷酸化也可以通过O-Glcnacylation（一种响应营养和压力来调节信号传导的PTM）进行竞争性调节。我们的目标是阐明这些PTM在病毒潜伏期中的功能，并定义其串扰在通过VP16反式激活中调节病毒蛋白的早期基因表达中的作用。这项工作的结果将确定转录因子或蛋白质修饰可能是未来开发用于HSV重新激活的治疗干预措施的靶标的酶。