Project 1 - Chromatin and the lytic/latent balance

项目 1 - 染色质和裂解/潜在平衡

• 批准号: 10686362
• 负责人: DAVID M. KNIPE
• 金额: $ 47.31万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-02 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10686362
• 关键词: Afferent Neurons; Alzheimer' s Disease; Antiviral Agents; Binding; Binding Sites; CCCTC-binding factor; Cells; Chromatin; Chromatin Structure; Collaborations; DNA; Disease; Epigenetic Process; Equilibrium; Euchromatin; Fibroblasts; Genes; Genetic Transcription; Genome; Goals; HIV; Herpes Simplex Infections; Herpesvirus 1; Heterochromatin; Histones; Human; Immediate-Early Genes; In Vitro; Induced pluripotent stem cell derived neurons; Infection; Knowledge; Latent virus infection phase; Location; Lytic; Lytic Phase; Maintenance; Medical; Modeling; Molecular Conformation; Mus; Mutation; Neurodegenerative Disorders; Neurons; Repression; Research; Research Project Grants; Risk; Role; Simplexvirus; Site; Structure; Structure of trigeminal ganglion; System; Testing; Therapeutic; Transcript; Viral; Viral Genome; Viral Physiology; Viral Proteins; Virus; Virus Latency; epigenetic regulation; epigenetic silencing; gene product; genital herpes; in vivo; induced pluripotent stem cell; inhibitor; latency associated transcript; latent infection; novel; novel strategies; novel therapeutic intervention; novel therapeutics; promoter; recruit; small molecule; three dimensional structure; transmission process

Project Summary/Abstract – Project 1 The long-term goals of the research of this project are 1) to define the mechanisms by which herpes simplex virus 1 (HSV-1) gene products promote euchromatin on viral lytic genes during lytic infection and heterochromatin on these genes during latent infection of sensory neurons to determine the switch between lytic and latent infection, and 2) to apply this knowledge to develop therapeutics for HSV latent infection. Antiviral drugs that inhibit HSV lytic infection have been developed, but there is no antiviral therapeutic for latent infection. Previously, we have defined many aspects of HSV latent infection of sensory neurons in murine trigeminal ganglia, in particular the structure of latent HSV-1 chromatin and the viral gene products that regulate viral chromatin structure. In the proposed research, we will test mechanisms of latent infection in the murine trigeminal ganglion model, in cultured mouse neurons and, for greater medical relevance, in human inducible pluripotent stem cell-derived neurons. We will focus on functions of viral gene products that promote a stable latent infection that is poised for reactivation. All aims are integrated with Projects 2 and 3 and Cores A and B. Specific Aim 1 will test various hypotheses about how the latency-associated transcript (LAT) promotes total histone and facultative heterochromatin loading on latent HSV-1 DNA. We will test whether LAT promotes epigenetic silencing by antisense transcription by inserting transcriptional terminators in the LAT transcriptional unit in the HSV-1 genome, and testing the effects on chromatin, transcripts antisense to ICP4, and expression of downstream antisense genes. We will test if LAT recruits histone loaders and epigenetic factors to the HSV-1 genome to promote loading of heterochromatin to the viral genome and stable maintenance of poised latency. Specific Aim 2 will explore mechanisms by which ICP0 promotes stable maintenance of the latent HSV-1 genome. We will determine the role of ICP0 in maintaining the viral genome, heterochromatin, and LAT expression in human neurons, murine neurons, and murine in vivo systems. The effects of a novel, exciting ICP0-specific inhibitory small molecule will be defined in murine and human neuronal latency systems. Specific Aim 3 will study the mechanism of action of the CCCTC-binding factor (CTCF) binding site (CTRL2) between the LAT and ICP0 gene promoters, which serves as a chromatin insulator and promotes reactivation. We will define the role of CTCF binding at the at the CTRL2 site on establishment and maintenance of latent infection. We will define the role of CTRL2 in the 3-dimensional structure of viral chromatin in human and murine neurons with Project 3. These studies will provide important new basic knowledge of the mechanisms of HSV latent infection and may enable new therapeutics for the HSV latent infection.

项目概要/摘要-项目1 该项目研究的长期目标是：1）确定疱疹病毒感染的机制， 单纯病毒1（HSV-1）基因产物在裂解感染期间促进病毒裂解基因上的常染色质， 在潜伏感染感觉神经元期间，这些基因上的异染色质，以确定 裂解和潜伏感染，和2）应用这些知识开发HSV潜伏感染的治疗剂。 已经开发了抑制HSV裂解性感染的抗病毒药物，但是对于HSV裂解性感染没有抗病毒治疗。 潜伏感染以前，我们已经确定了HSV潜伏感染感觉神经元的许多方面， 鼠三叉神经节，特别是潜伏的HSV-1染色质和病毒基因产物的结构， 调节病毒染色质结构。在拟议的研究中，我们将测试潜伏感染的机制， 小鼠三叉神经节模型，在培养的小鼠神经元中，为了更大的医学相关性，在人类中 诱导性多能干细胞衍生的神经元。我们将重点研究病毒基因产物的功能， 一种稳定的潜伏感染随时可能重新激活所有目标都与项目2和项目3以及核心相结合 A和B。 具体目标1将测试关于潜伏相关转录本（LAT） 促进总组蛋白和兼性异染色质加载在潜伏HSV-1 DNA上。我们将测试LAT是否 通过在LAT中插入转录终止子，通过反义转录促进表观遗传沉默 HSV-1基因组中的转录单位，并测试对染色质，ICP 4反义转录物， 和下游反义基因的表达。我们将测试LAT是否招募组蛋白装载器和表观遗传学 在HSV-1基因组中的特异性转录因子，以促进异染色质装载到病毒基因组中，并稳定 保持稳定的延迟。 具体目标2将探索ICP 0促进潜伏期的稳定维持的机制。 HSV-1基因组。我们将确定ICP 0在维持病毒基因组、异染色质和LAT中的作用 在人神经元、鼠神经元和鼠体内系统中表达。一部小说的效果，令人兴奋 将在鼠和人神经元潜伏系统中定义ICP 0特异性抑制性小分子。 具体目标3将研究CCCTC结合因子（CTCF）结合位点的作用机制 在LAT和ICP 0基因启动子之间存在CTRL 2，其作为染色质绝缘体并促进 重新激活我们将定义CTCF在CTRL 2位点结合的作用， 保持潜伏感染。我们将定义CTRL 2在病毒的三维结构中的作用， 染色质在人类和小鼠神经元与项目3。 这些研究将为HSV潜伏感染机制提供重要的新的基础知识 并可能为HSV潜伏感染提供新的治疗方法。