Investigation of herpes simplex virus -1 neurotropism in SCID DRG xenografts

SCID DRG 异种移植物中单纯疱疹病毒-1 向神经性的研究

• 批准号: 7638379
• 负责人: Ann Arvin
• 金额: $ 20.19万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-22 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7638379
• 关键词: Address; Afferent Neurons; Animal Model; Antiviral Agents; Attenuated; Axonal Transport; Benign; Biology; Cell Communication; Cells; Characteristics; Chemicals; Disease; Double Stranded DNA Virus; Encephalitis; Epithelial Cells; Evaluation; Event; Exhibits; Failure; G0 Phase; Ganglia; Gene Expression; Genes; Genetic Transcription; Genital system; Genome; Glycoproteins; Goals; Herpesviridae; Herpesvirus 1; Herpesvirus Type 3; Human; Human Herpesvirus 2; Immune system; Immunocompromised Host; Individual; Infection; Investigation; Lesion; Life; Lytic; Methods; Modeling; Molecular; Mucous Membrane; Mus; Mutation; Myxoid cyst; Nerve Fibers; Neuroglia; Neurons; Neuropathogenesis; Neurotropism; Oral; Oryctolagus cuniculus; PVRL1; Patients; Pattern; Peripheral Nerves; Pharmaceutical Preparations; Public Health; Recombinants; Replication Initiation; Rest; Reverse Transcriptase Polymerase Chain Reaction; Rodent Model; Sensory Ganglia; Severe Combined Immunodeficiency; Signal Pathway; Signal Transduction; Simplexvirus; Site; Skin; Specimen; Spinal Ganglia; Stimulus; Surface; System; Thymidine Kinase; Tissues; Transcript; United States; Vaccines; Viral; Viral Genes; Viral Genome; Viral Proteins; Virion; Virus; Virus Latency; Work; Xenograft Model; Xenograft procedure; afferent nerve; anterograde transport; cell type; gene function; high risk; high voltage electron microscopy; human disease; in vivo; insight; latency associated transcript; latent infection; mouse model; mutant; neonate; neuronal cell body; neurovirulence; prevent; protein expression; public health relevance; receptor; recombinant virus; research study; response; satellite cell; stressor; vaccine candidate; virus host interaction

DESCRIPTION (provided by applicant): Herpes simplex virus-1 (HSV-1) is human alphaherpesvirus that establishes a lifelong latent infection in peripheral nerve ganglia following primary infection. HSV-1 infections are generally benign, although its capacity for neurovirulence and neuroinvasiveness are the primary mechanisms through which HSV-1 can cause harmful disease in humans, especially in neonates and immunocompromised hosts. Our overall objective is to develop a model for examining HSV-1 neuropathogenesis in human sensory ganglia in vivo. We will evaluate HSV-1 infection of human dorsal root ganglion (DRG) xenografts in mice with severe combined immunodeficiency (SCID), exploiting the system that we created to investigate varicella- zoster virus (VZV) neuropathogenesis. The biology of HSV-1 infection is similar to VZV in that both HSV-1 and VZV establish latency within sensory ganglia following primary infection. Studies of VZV in the SCIDhu DRG model have provided the first opportunity to examine replication of a human alphaherpesvirus within cells that comprise human DRG in vivo. The DRG xenograft model has the potential to reveal characteristics of HSV-1 neuropathogenesis in the natural human host tissue microenvironment in vivo in an experimental system that will add substantially to observations from rodent models. Experiments will address three specific aims: (1) we will define the course of events that follows HSV-1 inoculation of human DRG xenografts in SCID mice, identifying what cell types within DRG are permissive for HSV-1 gene expression, whether neurons and/or satellite cells become productively infected and whether HSV-1 undergoes the pattern of transition to persistence in human neurons that we have observed in VZV-infected DRG xenografts; (2) we will investigate HSV-1 gene functions through the evaluation of recombinant HSV-1 strains, in particular we will examine the requirement for HSV-1 thymidine kinase (TK) during initial infection and persistence in DRG, and gD mutants for their capacity for viral entry; (3) if HSV-1 is shown to establish persistence in DRG xenografts, we will assess whether this model can be used to study HSV-1 reactivation by explanting latently-infected DRG xenografts and treating with agents that trigger neural cell signaling pathways and increase HSV-1 reactivation in rodent models. This work is intended to demonstrate the feasibility of using DRG xenografts in SCID mice to explore the molecular mechanisms of HSV-1 neuropathogenesis in differentiated human sensory neurons and non-neuronal cells within their sensory ganglia tissue microenvironment in vivo. In addition to new insights about basic virus-host interactions, such a model has potential value for studying antiviral drugs and live attenuated HSV-1 vaccine candidates to treat or prevent human disease caused by this common virus. PUBLIC HEALTH RELEVANCE: Herpes Simplex Virus 1 (HSV-1) causes oral and genital lesions and encephalitis. These infections remain an important public health problem in the United States. Serious complications from HSV-1 can occur in healthy people and in those who have diseases that impair their immune systems. Our goal is to develop a model to study how HSV-1 infects human nerve cells that will have potential value for developing new drugs and vaccines.

描述（由申请人提供）：单纯疱疹病毒-1 （HSV-1）是人类α疱疹病毒，在原发性感染后周围神经节中建立终身潜伏感染。1型单纯疱疹病毒感染通常是良性的，尽管其神经毒力和神经侵袭能力是1型单纯疱疹病毒在人类，特别是在新生儿和免疫功能低下的宿主中引起有害疾病的主要机制。我们的总体目标是建立一个在体内检测HSV-1在人类感觉神经节中的神经发病机制的模型。我们将利用我们创建的系统来研究水痘-带状疱疹病毒（VZV）的神经发病机制，评估人类背根神经节（DRG）异种移植物在严重联合免疫缺陷（SCID）小鼠中的HSV-1感染。HSV-1感染的生物学原理与VZV相似，即HSV-1和VZV在初次感染后都在感觉神经节内建立潜伏期。在SCIDhu DRG模型中对VZV的研究首次提供了在体内检测包含人类DRG的细胞内人类甲疱疹病毒复制的机会。DRG异种移植模型有可能在实验系统中揭示HSV-1在人体自然宿主组织微环境中的神经发病特点，这将大大增加啮齿动物模型的观察结果。实验将有三个具体目的：(1)我们将确定在SCID小鼠中接种HSV-1人类DRG异种移植物后的事件过程，确定DRG内哪些细胞类型允许HSV-1基因表达，神经元和/或卫星细胞是否会产生感染，以及HSV-1是否经历了我们在vzv感染的DRG异种移植物中观察到的人类神经元过渡到持久性的模式；(2)我们将通过评估重组HSV-1菌株来研究HSV-1基因的功能，特别是我们将研究DRG和gD突变体在初始感染和持续感染时对HSV-1胸苷激酶（TK）的需求，以确定它们的病毒进入能力；(3)如果HSV-1在DRG异种移植物中显示出持久性，我们将评估该模型是否可以用于研究HSV-1的再激活，方法是外植潜伏感染的DRG异种移植物，并在啮齿动物模型中使用触发神经细胞信号通路和增加HSV-1再激活的药物。本工作旨在证明在SCID小鼠中使用DRG异种移植的可行性，以探索HSV-1在人感觉神经节组织微环境中分化的人感觉神经元和非神经元细胞的神经发病分子机制。除了对基本病毒-宿主相互作用的新见解外，该模型还具有研究抗病毒药物和减毒HSV-1活疫苗候选物以治疗或预防由这种常见病毒引起的人类疾病的潜在价值。公共卫生相关性：单纯疱疹病毒1 （HSV-1）引起口腔和生殖器病变和脑炎。这些感染在美国仍然是一个重要的公共卫生问题。1型单纯疱疹病毒的严重并发症可发生在健康人群和免疫系统受损疾病患者身上。我们的目标是开发一个模型来研究1型单纯疱疹病毒如何感染人类神经细胞，这将对开发新药和疫苗具有潜在价值。