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

• 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
• 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

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型和2型；HSV-1和HSV-2）是世界上最常见的病毒感染之一，影响超过70%的人口。典型的α疱疹病毒感染会导致周围神经系统（PNS）神经节终身潜伏。潜伏的病毒基因组可以重新激活，引发播散性感染，通常导致疱疹水疱、生殖器病变或角膜炎。原发感染始于粘膜表面上皮细胞的生产性感染。复制后，子代病毒颗粒主要侵入背根和三叉神经节（DRG和TG）以及其他自主交感神经节（例如SCG）建立潜伏期。感染的潜伏期、重新激活和随后的传播的建立受到许多细胞内在的、组织特异性的和系统性因素的影响，这些因素很难剖析。现有的抗疱疹药物以病毒 DNA 复制为目标，但对潜伏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析潜伏和重新激活的分子机制，以便能够设计新的疗法。目前使用的 HSV-1 潜伏期神经元培养模型采用分离的啮齿动物原代神经元，并使用阿昔洛韦抑制 DNA 复制并迫使感染进入潜伏期。这并不理想，不仅因为病毒粒子直接感染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可能无法进一步复制。最近，我们通过在不使用药物的情况下用低浓度的兽医病原体伪狂犬病病毒感染隔离神经元的分离轴突，开发了一种可重复且可重新激活的体外潜伏系统。由于在这些条件下感染的结果总是沉默的，因此我们研究了能够逃脱基因组沉默的机制。我们发现了两种不同的途径可以防止 PRV 基因组沉默。我们将开发这个系统来探索人类病原体 HSV-1 潜伏建立和重新激活的分子机制。识别能够摆脱沉默的神经元应激途径将有助于设计新的治疗策略来控制复发性疱疹病毒再激活患者的病毒再激活和相关病理。

项目成果

Cell Intrinsic Determinants of Alpha Herpesvirus Latency and Pathogenesis in the Nervous System.

阿尔法疱疹病毒潜伏期和神经系统发病机制的细胞内在决定因素。

• 发表时间: 2023-11-22
• 作者: Salazar, Stephanie;Luong, Khanh T Y;Koyuncu, Orkide O
• 通讯作者: Koyuncu, Orkide O

Interferon-λ Activates a Differential Response in Peripheral Neurons That Is Effective against Alpha Herpesvirus Infections.

干扰素-α 激活周围神经元的差异反应，可有效对抗阿尔法疱疹病毒感染。

• 发表时间: 2023-09-07
• 作者: Salazar, Stephanie;Luong, Khanh T Y;Nua, Taulima;Koyuncu, Orkide O
• 通讯作者: Koyuncu, Orkide O