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：
8678830
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 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8678830
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感染过单纯疱疹病毒。在最初的裂解性感染后，HSV 在感觉神经元内建立永久的潜伏感染。潜伏病毒的重新激活不仅会导致病毒性疾病（新感染、失明和脑炎），还会导致艾滋病毒感染、糖尿病、心血管和神经退行性疾病。目前还没有有效的疫苗，也没有治疗可以消除潜伏期或防止重新激活。该项目的长期目标是通过定义控制 HSV-1 潜伏期建立和重新激活的机制来找到针对复发性 HSV 发作的干预措施。 HSV-1 裂解性感染期间的基因表达级联始于病毒体蛋白 VP16 与宿主因子 Oct-1 和 HCF-1 激活立即早期 (IE) 基因转录。相比之下，重新激活中的初始事件延迟的影响仍然不明确。该提议的中心假设是VP16表达和活性的调节是潜伏期建立和从潜伏期重新激活的基础。这两个级别的控制涉及多个正负输入，以允许或抑制体内感觉神经元中的病毒复制。目标 1. 该项目将确定体内感觉神经元中 VP16 基因从头激活和沉默的机制。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反应性启动子元件以及相应转录因子的作用进行调节，以允许或抑制初始感染或退出潜伏期时的裂解复制。在小鼠感染和潜伏期模型中使用重组病毒，我们发现 VP16 的表达对于触发潜伏期的退出是必要且充分的，并且我们鉴定了 VP16 基因的神经元特异性启动子。我们将测试该启动子是否在急性感染期间和重新激活期间控制进入裂解期感染。我们将确定预测的转录因子是否与该启动子的各个元件结合，以正向或负向调节 VP16 基因表达，无论是单独、组合还是竞争。目标 2. 该项目将定义 VP16 共激活剂相互作用对于 VP16 依赖性退出潜伏期至关重要，并确定体内调节这些相互作用的机制。我们的数据强烈表明，HSV 的潜伏期退出受到 CK2 介导的磷酸化的调节，并且这种磷酸化也可能受到 O-GlcNAcylation（一种调节响应营养和应激信号的 PTM）的竞争性调节。我们的目标是阐明这些 PTM 在病毒潜伏期中的功能，并确定它们的串扰通过 VP16 反式激活调节病毒蛋白立即早期基因表达的作用。这项工作的结果将确定转录因子或蛋白质修饰这些酶可能成为未来开发 HSV 重新激活治疗干预措施的目标。