Role of DNA damage in the early steps of HSV infection and latency in neurons

DNA 损伤在 HSV 感染早期阶段和神经元潜伏期中的作用

• 批准号: 8895429
• 负责人: Matthew D. Weitzman
• 金额: $ 45.23万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8895429
• 关键词: Ablation; Acyclovir; Address; Afferent Neurons; Animal Model; Antiviral Agents; Cell Nucleus; Cells; DNA; DNA Damage; DNA Viruses; DNA repair protein; Data; Dependence; Deposition; Disease; Encephalitis; Environment; Episome; Epithelial Cells; Fingers; Gene Expression; Genes; Genetic; Genome; Goals; Herpesviridae; Herpesvirus 1; Human; Immediate-Early Proteins; Immunohistochemistry; In Situ Hybridization; Infection; Keratitis; Knowledge; Lead; Link; Lytic; Lytic Phase; Maps; Molecular; Mus; Nervous system structure; Neuroglia; Neurons; Nucleosomes; Oligonucleotide Microarrays; Outcome; Peripheral Nervous System; Pharmaceutical Preparations; Positioning Attribute; Presynaptic Terminals; Process; Productivity; Proteins; Recurrence; Recurrent disease; Regulation; Role; Set protein; Signal Transduction; Simplexvirus; Site; Small Interfering RNA; Staging; Stress; Structure of trigeminal ganglion; Surface; Techniques; Therapeutic; Viral; Viral Genome; Viral Proteins; Virus; Virus Diseases; Work; base; behavioral response; cellular targeting; human DNA; in vivo; inhibitor/antagonist; insight; latent infection; lytic replication; mouse model; mutant; novel; pathogen; prevent; protein expression; public health relevance; reactivation from latency; response; stressor; ubiquitin ligase; ubiquitin-protein ligase; ultraviolet damage; virus host interaction

DESCRIPTION (provided by applicant): Herpes Simplex Virus type 1 (HSV-1) is a human DNA virus that causes a number of diseases involving the human nervous system, including keratitis and encephalitis. The virus is characterized by its ability to form latent infections in neurons of the peripheral nervous system, from which it can reactivate to cause recurrent disease. Our long-term goal is to understand the cellular responses that contribute to establishment of HSV-1 latency specifically in neurons. The latent genome persists in an episomal chromatinized state for the lifetime of the host without detectable protein expression. Reactivation can occur spontaneously or be induced by various forms of stress. The viral immediate early protein ICP0 is a multifunctional protein important for stimulating the initiation f the lytic cycle and also efficient reactivation of latent or quiescent genomes. Exactly how ICP0 creates a favorable cellular environment during lytic replication and promotes reactivation from latency remains unclear, but dependence on the E3 ubiquitin ligase activity of the RING finger domain in ICP0 suggests that degradation of its substrates is involved. Our labs and others have shown that HSV-1 infection activates and exploits aspects of the cellular DNA damage signaling cascade. We have also shown that the DNA damage machinery can act as an intrinsic cellular defense that is inactivated through degradation of specific DNA repair proteins by ICP0. Our central hypothesis is that both the DNA damage response and the behavior of ICP0 are different in neuronal cells, and that these two factors contribute to establishment of latency specifically in neurons. The objective of this proposal is to define the role of the DNA damage response in circularization and nucleosome deposition on the incoming viral genome at the earliest stages of infection, and to determine how this impacts the decision to form lytic or laten HSV infections in neurons. We will examine how ICP0 controls these processes and will determine why the ICP0 protein does not accumulate in the nucleus of infected neurons. The approach will employ novel techniques to determine how cellular responses impact infection in human neurons in culture and in animal models. These neurovirological studies are significant because they are expected to provide insights into virus-host interactions during HSV-1 infection and the early responses that determine the outcome of infection in neurons and non-neuronal cells. Knowledge of HSV-1 neuronal latency will suggest novel targets for developing antivirals against this significant human pathogen.

描述（由申请人提供）： 1 型单纯疱疹病毒（HSV-1）是一种人类 DNA 病毒，可引起多种涉及人类神经系统的疾病，包括角膜炎和脑炎。该病毒的特点是能够在周围神经系统的神经元中形成潜伏感染，并可以重新激活以引起复发性疾病。我们的长期目标是了解有助于建立 HSV-1 潜伏期（特别是在神经元中）的细胞反应。潜在基因组在宿主的一生中持续处于游离染色质状态，而没有可检测到的蛋白质表达。重新激活可以自发发生或由各种形式的压力诱导。病毒立即早期蛋白 ICP0 是一种多功能蛋白，对于刺激裂解周期的启动以及潜在或静止基因组的有效重新激活非常重要。 ICP0 究竟如何在裂解复制过程中创造有利的细胞环境并促进潜伏期重新激活尚不清楚，但 ICP0 中环指结构域对 E3 泛素连接酶活性的依赖表明，其中涉及其底物的降解。我们的实验室和其他实验室已经表明，HSV-1 感染会激活并利用细胞 DNA 损伤信号级联的各个方面。我们还表明，DNA 损伤机制可以作为一种内在的细胞防御，通过 ICP0 降解特定的 DNA 修复蛋白来使其失活。我们的中心假设是，神经元细胞中的 DNA 损伤反应和 ICP0 的行为都是不同的，并且这两个因素有助于神经元中特异潜伏期的建立。该提案的目的是确定感染早期阶段 DNA 损伤反应在传入病毒基因组上的环化和核小体沉积中的作用，并确定这如何影响神经元中形成裂解性或潜伏性 HSV 感染的决定。我们将研究 ICP0 如何控制这些过程，并确定为什么 ICP0 蛋白不会在受感染神经元的细胞核中积聚。该方法将采用新技术来确定细胞反应如何影响培养物和动物模型中人类神经元的感染。这些神经病毒学研究意义重大，因为它们有望深入了解 HSV-1 感染期间病毒与宿主的相互作用，以及决定神经元和非神经元细胞感染结果的早期反应。对 HSV-1 神经元潜伏期的了解将为开发针对这种重要人类病原体的抗病毒药物提供新的靶标。