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）每年在全球数十亿儿童中引起水痘（水痘），之后病毒潜伏在沿着整个神经轴的神经节神经元中。几十年后，VZV特异性细胞介导的免疫力下降，病毒重新激活，导致带状疱疹（带状疱疹），其特征是疼痛和皮疹仅限于1-3个皮区。带状疱疹每年影响约100万美国人，并且通常并发慢性神经根疼痛（带状疱疹后神经痛，PHN）和其他严重的神经系统疾病（小脑炎，肌病和血管病）和眼部疾病，这些疾病共同引起 瘫痪、失明和死亡到2030年，22%的美国人（6500万人）将超过65岁，85岁以上的人口将增加两倍，达到800万。带状疱疹的发病率和严重程度从 随着年龄的增长，VZV特异性免疫力自然下降，在器官移植受者和癌症及AIDS患者中观察到更严重的宿主免疫缺陷。带状疱疹疫苗可在3年内将带状疱疹的发病率降低50%，但即使对60岁以上的人进行免疫接种，每年仍有500，000例带状疱疹和200，000例PHN。因此，随着老年人口的增加，带状疱疹及其伴随的神经和眼部并发症的发病率也增加。在1983年首次证明VZV在人类神经节中潜伏之后，我们随后对来自>900名随机尸检受试者的> 7，000个人类神经节的分析揭示了整个病毒基因组在神经元中的存在，最有可能保持为具有可变VZV DNA丰度的组蛋白包被的附加体，VZV转录限于ORF 63，以及VZV基因表达的表观遗传调控。为了更精确地定义潜伏VZV基因组上的病毒基因表达和表观遗传标记的程度，我们检查了VZV感染的分化神经元，这些神经元在培养中保持活力长达3周，没有细胞病变效应;用干扰素治疗这些神经元显著降低了病毒基因表达。因此，我们假设，神经元治疗，以限制VZV基因的表达将产生一个模型系统，概括VZV转录潜伏感染的人类神经节神经元。为了验证我们的假设，我们将：在体外用抗病毒剂和由先天性和适应性免疫系统的细胞产生的分子处理VZV非溶解性感染的神经元，并通过RT-PCR检测VZV基因表达（目的1）;确定在用阿昔洛韦和多种细胞因子处理后，非裂解性感染的神经元内VZV DNA、VZV基因表达和VZV基因组上存在的表观遗传标记的物理状态（目的2）。在体外开发VZV潜伏期模型将允许通常需要获得数千个人类神经节的研究。深入了解VZV如何长期存在于潜伏感染的人类神经细胞中是制定预防导致病毒再激活的级联事件的策略的先决条件，这是严重神经系统疾病和失明的原因，特别是在迅速增加的老年人和免疫功能低下的人群中。