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.

项目摘要/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，最多存在90％ 世界人口。 HSV-1在外围神经节中以潜在感染的形式持续生命 支配头部和颈部。潜在感染的HSV-1重新激活与显着有关 发病率，包括可能表现为结膜炎，睑缘炎或角膜上皮的疱疹眼病 和基质角膜炎。病毒的复发性眼重新激活会导致复发性疱疹性角膜炎，这 仍然是发达国家传染失明的主要原因。因此，有必要 了解该病毒在神经元中如何保持潜在，以最终防止重新激活和复发感染。 在潜伏期期间，病毒基因组与反射核结构有关，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 已知。我已经确定了一种细胞蛋白ATRX，这对于维持HSV-1潜伏期至关重要。在目标1中 这项建议，我将利用一种新型的HSV-1潜伏期主要神经元模型，该模型允许建立一个 在微流体腔中培养的周围神经元中的潜在感染。我将测试ATRX的假设 涉及保留HSV-1潜伏期并检查ATRX如何保持的关键细胞蛋白 基于异染色质基因沉默。此外，我将利用一种眼感染模型来了解 ATRX耗竭在体内的影响。这些研究将提供有关机械的见解 基于异染色质的沉默在保持HSV-1潜伏期和防止复发的重新激活方面。 病毒基因组共定位不同的神经元水平的异质性 核域，可能导致不同类型的延迟，或多或少容易重新激活。我 已经发现神经元暴露于I型干扰素（IFN）导致形成反射性PML- 核体，以及在没有I型IFN的情况下建立的潜伏期更受限制的延迟。 因此，在本提案的目标2中，我将研究PML核体如何响应I型IFN 并检查PML核体在压实潜在的HSV-1基因组并使它们的贡献 对重新激活的难治。这些目的的最终目标是了解如何保留长期HSV-1 通过维持ATRX与病毒基因组的关联和/或促进HSV-的压实来进行潜伏期 反射性PML核体中的1个基因组。