Neurobiology of Varicella Zoster Virus

水痘带状疱疹病毒的神经生物学

• 批准号: 8979278
• 负责人: DONALD GILDEN
• 金额: $ 32.66万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8979278
• 关键词: 2' ,3' -Cyclic-Nucleotide Phosphodiesterases; Acquired Immunodeficiency Syndrome; Acyclovir; Affect; Aftercare; Age; American; Antiviral Agents; Autopsy; Biological Assay; Biological Models; Blindness; Cells; Cellular Immunity; Cessation of life; Chickenpox; Child; Chronic; DNA Viruses; Disease; Elderly; Epigenetic Process; Episome; Event; Exanthema; Ganglia; Gene Expression; Gene Expression Profile; Genetic Transcription; Glial Fibrillary Acidic Protein; Herpes zoster disease; Herpesvirus Type 3; Histones; Hour; Human; Immune; Immune system; Immunity; Immunization; Immunocompromised Host; In Vitro; Incidence; Individual; Infection; Interferons; Laboratories; Malignant Neoplasms; MicroRNAs; Modeling; Myelitis; Negative Staining; Neuraxis; Neurobiology; Neurologic; Neurons; Organ Transplantation; Pain; Paralysed; Patients; Population; Postherpetic neuralgia; Reverse Transcriptase Polymerase Chain Reaction; Severities; Staining method; Stains; Stroke; Testing; Time; Transcript; Transplant Recipients; Tubulin; Vaccinated; Vaccines; Vascular Diseases; Viral Genes; Viral Genome; Virus; Virus Latency; base; chromatin immunoprecipitation; chronic pain; cytokine; deep sequencing; epigenetic marker; epigenetic regulation; killings; physical state; prevent; public health relevance; tissue culture; viral DNA

 DESCRIPTION (provided by applicant): Varicella zoster virus (VZV) causes varicella (chickenpox) in billions of children worldwide annually, after which virus becomes latent in ganglionic neurons along the entire neuraxis. Decades later, VZV-specific cell-mediated immunity declines and virus reactivates, resulting in zoster (shingles), characterized by pain and rash restricted to 1-3 dermatomes. Zoster affects ~1 million Americans yearly and is often complicated by chronic radicular pain (postherpetic neuralgia, PHN) and other serious neurological (cerebellitis, myelitis and vasculopathy) and ocular disorders that collectively cause paralysis, blindness and death. By 2030, 22% of Americans (65 million people) will be >age 65, and the >85 population will triple to >8 million. The incidence and severity of zoster range from a natural decline in VZV-specific immunity with advancing age to more serious host immune deficits seen in organ transplant recipients and in patients with cancer and AIDS. Zoster vaccine reduces the incidence of zoster by 50% for up to 3 years, yet even immunization of everyone >age 60 still predicts 500,000 cases of zoster and 200,000 cases of PHN annually. Thus, as the elderly population increases, so does the incidence of zoster and its attendant neurological and ocular complications. After first proving in 1983 that VZV is latent in human ganglia, our subsequent analyses of >7,000 human ganglia from >900 randomly autopsied subjects revealed the presence of the entire virus genome in neurons, most likely maintained as a histone-coated episome with variable VZV DNA abundance, VZV transcription restricted to ORF63, and epigenetic regulation of VZV gene expression. To more precisely define the extent of viral gene expression and epigenetic markers on the latent VZV genome, we examined VZV-infected differentiated neurons, which remained viable in culture up to 3 weeks without a cytopathic effect; treatment of these neurons with interferon- ¿ significantly reduced viral gene expression. Thus, we hypothesize that neurons treated to limit VZV gene expression will yield a model system that recapitulates VZV transcription in latently infected human ganglionic neurons. To test our hypothesis, we will: treat VZV non-lytically infected neurons in vitro with antiviral agents and molecules produced by cells of the innate and adaptive immune systems and examine VZV gene expression by RT-PCR (Aim 1); determine the physical state of VZV DNA, VZV gene expression and epigenetic markers present on VZV genomes within non-lytically infected neurons after treatment with acyclovir and multiple cytokines (Aim 2). Developing a model of VZV latency in vitro will allow studies that would normally require acquisition of thousands of human ganglia. An in-depth understanding of how VZV resides long-term in latently infected human nerve cells is prerequisite to developing strategies that prevent the cascade of events leading to virus reactivation, a cause of serious neurologic disease and blindness, particularly in the rapidly increasing elderly and immunocompromised populations.

 描述（由申请人提供）：水痘带状疱疹病毒（VZV）每年在全世界数十亿儿童中引起水痘（水痘），此后病毒潜伏在整个神经轴的神经节神经元中，数十年后，VZV 特异性细胞介导的免疫力下降，并且导致水痘。病毒重新激活，导致带状疱疹（带状疱疹），其特征是仅限 1-3 个皮区出现疼痛和皮疹 带状疱疹影响约 100 万人。美国人每年都患有慢性根性疼痛（带状疱疹后神经痛，PHN）和其他严重的神经系统疾病（小脑炎、脊髓炎和血管病变）和眼部疾病，这些疾病共同导致 到 2030 年，22% 的美国人（6500 万人）年龄将超过 65 岁，而 85 岁以上的人口将增加三倍，达到 800 万人以上。 随着年龄的增长，VZV 特异性免疫力会自然下降，从而导致器官受体以及癌症和艾滋病患者出现更严重的宿主免疫缺陷，在长达 3 年内，带状疱疹的发病率会降低 50%，但即使每个人都进行了免疫。 60 岁以上的人预计每年将有 500,000 例带状疱疹和 200,000 例 PHN。因此，随着老年人口的增加，带状疱疹和 PHN 的发病率也会增加。在 1983 年首次证明 VZV 潜伏在人类神经节中后，我们对来自超过 900 名随机尸检受试者的超过 7,000 个人类神经节进行了分析，揭示了神经元中存在整个病毒基因组，很可能是作为一个病毒基因组存在的。组蛋白包被的附加体具有可变的 VZV DNA 丰度、VZV 转录仅限于 ORF63 以及 VZV 基因表达的表观遗传调控，以更精确地定义病毒基因表达和表观遗传的程度。通过对潜伏 VZV 基因组上的标记进行检测，我们检测了感染 VZV 的分化神经元，这些神经元在培养中可存活长达 3 周，而不会出现细胞病变效应；因此，我们帮助限制 VZV 基因表达的神经元产生了一个模型系统，该系统可以在潜伏感染的人类神经节神经元中重现 VZV 转录。为了检验我们的假设，我们将：在体外使用抗病毒剂和先天性和适应性免疫系统细胞产生的分子，并通过 RT-PCR 检查 VZV 基因表达（目标 1）确定 VZV DNA 的物理状态、VZV 基因表达；使用阿昔洛韦和多种细胞因子治疗后，非裂解性感染神经元内的 VZV 基因组上存在表观遗传标记（目标 2），开发 VZV 潜伏期体外模型将允许通常需要采集数千个人类神经节的研究。深入了解 VZV 如何在潜伏感染的人类神经细胞中长期驻留，是制定策略的先决条件，以防止导致病毒再激活的级联事件，病毒再激活是严重神经系统疾病的原因之一。失明，特别是在迅速增加的老年人和免疫功能低下的人群中。