Establishing a Laboratory Model of Herpes simplex virus (HSV) Latency Using Compartmented Neuron Cultures

使用区室神经元培养物建立单纯疱疹病毒 (HSV) 潜伏期的实验室模型

基本信息

批准号：
10326688
负责人：
Orkide O. Koyuncu
金额：
$ 23.55万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-15 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10326688
关键词：
Acyclovir Adult Affect Animal Model Antiviral Agents Architecture Axon Biological Models Bulla Capsid Cell Nucleus Cells Color DNA biosynthesis Decision Making Drug Targeting Drug usage Encephalitis Epithelial Cells Event Foundations Future Ganglia Genital Genitalia Genome Herpes Labialis Herpesviridae Herpesviridae Infections Herpesvirus 1 Herpesvirus Type 3 Human Human Herpesvirus 2 In Vitro Infection Injury Invaded Keratitis Laboratories Lesion Life Mammals Methodology Methods Modeling Molecular Monitor Motor Neurons Mucous Membrane Neurons Outcome Pathology Pathway interactions Patients Peripheral Nervous System Pharmaceutical Preparations Population Primary Infection Prodrugs Recurrence Regulation Reporter Reproducibility Rodent Signal Transduction Simplexvirus Stimulus Stress Structure of superior cervical ganglion Structure of trigeminal ganglion Surface Sympathetic Ganglia System Testing Tissues Viral Viral Genome Virion Virus Virus Diseases Virus Latency Virus Replication biological adaptation to stress cell motility design experimental study genital herpes guanine analog human pathogen in vitro Model inhibitor/antagonist latent infection mad itch virus nerve supply neuronal cell body neurotransmission novel novel therapeutic intervention novel therapeutics particle pathogen prevent reactivation from latency response to injury spinal nerve posterior root targeted agent trafficking viral DNA

项目摘要

Alpha herpesvirus infections (e.g. herpes simplex virus 1 and 2; HSV-1 and HSV-2) are among the most common virus infections in the world affecting more than 70% of human population. A typical alpha herpesvirus infection results in life-long latency in the peripheral nervous system (PNS) ganglia. Latent viral genomes can reactivate to start a disseminated infection often leading to herpes blisters, genital lesions or keratitis. The primary infection starts with a productive infection in the epithelial cells of mucosal surfaces. After replication, progeny virus particles mainly invade the dorsal root and trigeminal ganglia (DRG and TG), and other autonomic sympathetic ganglia (e.g. SCG) to establish latency. Establishment of latency, reactivation, and subsequent spread of infection is affected by many cell intrinsic, tissue-specific and systemic factors that are challenging to dissect. The available anti-herpetic drugs target viral DNA replication but have no effect on latent or reactivating viral genomes. Consistent and reproducible laboratory models are required to dissect the molecular mechanisms of latency and reactivation to be able to design novel therapies. Currently used neuron culture models of HSV-1 latency employs dissociated rodent primary neurons with the use of acyclovir to inhibit DNA replication and force the infection into latency. This is not ideal, not only because the virions are directly infecting neuronal cell bodies instead of axons, but also because the viral DNA that incorporated the guanine analogue might not be competent for further replication. Recently, we have developed a reproducible and reactivatable in vitro latency system by infecting isolated axons of compartmented neurons with a low concentration of the veterinary pathogen, pseudorabies virus without the use of drugs. Since the outcome of infection is always silenced under these conditions, we studied mechanisms that enable escape from genome silencing. We discovered two distinct pathways that prevented PRV genome silencing. We will develop this system to explore the molecular mechanisms of latency establishment and reactivation of the human pathogen, HSV-1. Identification of neuronal stress pathways enabling escape from silencing will contribute to designing novel therapeutic strategies to control virus reactivation and related pathologies in patients with recurrent herpesvirus reactivation.

α疱疹病毒感染（例如，单纯疱疹病毒1和2; HSV-1和HSV-2）是世界上最常见的病毒感染之一，影响了70％以上的人群。典型的α疱疹病毒感染导致周围神经系统（PNS）神经节的终生潜伏期。潜在病毒基因组可以重新激活，以开始传播感染，通常导致疱疹水泡，生殖器病变或角膜炎。主要感染始于粘膜表面上皮细胞中的生产性感染。复制后，后代病毒颗粒主要侵入背根和三叉神经节（DRG和TG），以及其他自主性交感神经节（例如SCG）以建立潜伏期。建立潜伏期，重新激活和随后的感染传播受许多细胞内在的，组织特异性和全身因素的影响，这些因素具有挑战性。可用的抗疱疹药物靶向病毒DNA复制，但对潜在或重新激活的病毒基因组没有影响。需要一致且可重复的实验室模型来剖定潜伏期和重新激活的分子机制，以设计新型疗法。当前使用的HSV-1潜伏期的神经元培养模型利用使用acyclovir来抑制DNA复制并将感染迫使潜伏期。这不是理想的，不仅是因为病毒体直接感染了神经元细胞体而不是轴突，还因为掺入鸟嘌呤类似物的病毒DNA可能无法胜任进一步复制。最近，我们通过感染具有低浓度的兽医病原体的分离式神经元的隔离轴突，开发了一种可再现和可重复的体外潜伏系统，而无需使用药物。由于在这些条件下总是沉默感染的结果，因此我们研究了能够摆脱基因组沉默的机制。我们发现了两种阻止PRV基因组沉默的不同途径。我们将开发该系统来探索人类病原体HSV-1的潜伏期建立和重新激活的分子机制。从沉默中逃脱的神经元应力途径的鉴定将有助于设计新型的治疗策略，以控制复发性疱疹病毒重新激活患者的病毒重新激活和相关病理。