The role of repressive nuclear bodies in latent herpes simplex virus infection

抑制核体在潜伏单纯疱疹病毒感染中的作用

• 批准号: 10192728
• 负责人: JON Blackburn SUZICH
• 金额: $ 5.1万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10192728
• 关键词: ATRX gene; Acute Promyelocytic Leukemia; Afferent Neurons; Antiviral Agents; Axon; Blepharitis; Blindness; Cells; Cellular Structures; Chemistry; Chromatin Structure; Conjunctivitis; Cornea; Data; Development; Disease; Environment; Epithelial Cells; Equilibrium; Experimental Models; Exposure to; Eye Infections; Eye diseases; Foundations; Future; Ganglia; Gene Silencing; Genetic Transcription; Genome; Goals; Head and neck structure; Herpesviridae; Herpesvirus 1; Herpetic Keratitis; Heterochromatin; Heterogeneity; Human; Image; Immune response; In Vitro; Infection; Inflammatory Response; Interferon Type I; Interferons; Keratitis; Knock-out; Lead; Life; Long-Term Effects; LoxP-flanked allele; Lytic Phase; Maintenance; Microfluidics; Modeling; Morbidity - disease rate; Mus; Neuroglia; Neurons; Nuclear; Nuclear Structure; Peripheral; Phase; Physicians; Play; Population; Proteins; Recurrence; Recurrent disease; Refractory; Resolution; Role; Scientist; Simplexvirus; Stimulus; Testing; Viral Genes; Viral Genome; Virus; Virus Diseases; Virus Shedding; Western World; base; career; clinically relevant; corneal epithelium; corneal scar; experience; histone modification; human pathogen; in vivo; in vivo Model; insight; knock-down; latent infection; lytic replication; neuronal cell body; novel; pathogen; preservation; prevent; recurrent infection; response; therapeutic target; transmission process; virology

PROJECT SUMMARY/ABSTRACT Herpes simplex virus type 1 (HSV-1) is a ubiquitous human pathogen that is present in up to 90% of the world’s population. HSV-1 persists for life in the form of a latent infection in the peripheral ganglia that innervate the head and neck. Reactivation of HSV-1 from latent infection is associated with significant morbidity, including herpetic eye disease which can manifest as conjunctivitis, blepharitis or corneal epithelial and stromal keratitis. Recurrent ocular reactivation of the virus can result in recurrent herpetic keratitis, which remains the leading cause of infectious blindness in the developed world. Therefore, there is a need to understand how the virus remains latent in neurons to ultimately prevent reactivation and recurrent infection. During latency, the viral genome is associated with repressive nuclear structures and is assembled into heterochromatin. The role of cellular heterochromatin-associated proteins in maintaining HSV-1 latency is not known. I have identified one cellular protein, ATRX, that is essential for maintaining HSV-1 latency. In Aim 1 of this proposal, I will utilize a novel primary neuronal model of HSV-1 latency that permits the establishment of a latent infection in peripheral neurons cultured in microfluidic chambers. I will test the hypothesis that ATRX is the key cellular protein involved in preserving HSV-1 latency and examine how ATRX maintains heterochromatin-based gene silencing. In addition, I will utilize an ocular model of infection to understand the effects of ATRX depletion in vivo. These studies will provide mechanistic insights into the contribution of heterochromatin-based silencing in maintaining HSV-1 latency and preventing recurrent reactivation. There is considerable heterogeneity at the neuronal level in the co-localization of viral genomes with different nuclear domains, likely resulting in different types of latency that are more or less susceptible to reactivation. I have found that exposure of neurons to type I interferon (IFN) results in the formation of repressive PML- nuclear bodies, as well as a more restricted form of latency than that established in the absence of type I IFN. Therefore, in Aim 2 of this proposal, I will investigate how PML-nuclear bodies form in response to type I IFN and examine the contribution of PML-nuclear bodies in compacting latent HSV-1 genomes and making them refractory to reactivation. The ultimate goal of these aims is to understand how to preserve long-term HSV-1 latency through either maintaining ATRX association with viral genomes and/or promoting compaction of HSV- 1 genomes in repressive PML-nuclear bodies.

项目总结/摘要 单纯疱疹病毒1型（HSV-1）是一种普遍存在的人类病原体，存在于高达90%的人中， 世界人口。HSV-1在外周神经节中以潜伏感染的形式持续终生， 使头部和颈部受神经支配。HSV-1从潜伏感染中的再活化与显著的 发病率，包括疱疹性眼病，可表现为结膜炎、睑缘炎或角膜上皮炎 和基质角膜炎。病毒的反复眼部再活化可导致复发性疱疹性角膜炎， 仍然是发达国家传染性失明的主要原因。因此，有必要 了解病毒如何在神经元中保持潜伏状态，以最终防止再激活和复发感染。 在潜伏期，病毒基因组与抑制性核结构相关，并组装成 异染色质细胞异染色质相关蛋白在维持HSV-1潜伏期中的作用不是 知道的我已经确定了一种细胞蛋白，ATRX，这是维持HSV-1潜伏期所必需的。目标1， 在这个提议中，我将利用一种新的HSV-1潜伏期的初级神经元模型， 在微流控室中培养的外周神经元的潜伏感染。我将检验ATRX是 关键的细胞蛋白参与保持HSV-1潜伏期，并检查ATRX如何维持 异染色质基因沉默此外，我将利用眼部感染模型来了解 体内ATRX耗竭的影响。这些研究将提供机械的见解的贡献， 基于异染色质的沉默在维持HSV-1潜伏期和防止复发性再激活中的作用。 在神经元水平上，具有不同基因的病毒基因组的共定位存在相当大的异质性。 核域，可能导致不同类型的潜伏期，或多或少容易被重新激活。我 发现神经元暴露于I型干扰素（IFN）导致抑制性PML的形成- 核小体，以及一个更有限的形式的潜伏期比建立在没有I型IFN。 因此，在本建议的目标2中，我将研究PML核体如何响应I型IFN而形成 并研究PML核体在压缩潜伏的HSV-1基因组并使其 难再活化的。这些目标的最终目标是了解如何长期保存HSV-1 通过维持ATRX与病毒基因组的结合和/或促进HSV-1的致密化来潜伏。 1抑制性PML核小体中的基因组。