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。

• 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
• 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

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）：世界上大多数人都感染过单纯疱疹病毒。在最初的溶解性感染之后，单纯疱疹病毒在感觉神经元内建立永久性潜伏感染。潜伏病毒的再激活不仅会导致病毒性疾病（新感染、失明和脑炎），而且还会导致艾滋病毒感染、糖尿病、心血管和神经退行性疾病。没有有效的疫苗可用，也没有治疗方法可以消除潜伏期或防止再激活。该项目的长期目标是通过定义控制HSV-1潜伏期的建立和再激活的机制，找到对复发性HSV发作的干预措施。病毒粒子蛋白VP16与宿主因子Oct-1和HCF-1激活即时早期（IE）基因转录，开始了HSV-1裂解感染过程中的基因级联表达。相反，初始事件在重新激活