A new in vitro neuron model of axonal transport and persistence of varicella zost

水痘带状疱疹轴突运输和持续性的新体外神经元模型

基本信息

批准号：
8606907
负责人：
Paul R. Kinchington
金额：
$ 16.86万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8606907
关键词：
Address Adult Affect Afferent Neurons Age Animal Model Antiviral Agents Area Axon Axonal Transport Capsid Cell Nucleus Chickenpox Chronic DNA Disease Dissection Effectiveness Event Gene Deletion Gene Expression Genes Genome Goals Growth Factor Herpes zoster disease Herpesviridae Herpesvirus Type 3 Histocompatibility Testing Human In Situ In Vitro Infection Intractable Pain Kinetics Label Libraries Life Medical Microfluidics Modeling Molecular Profiling Morbidity - disease rate Mutate Nature Neurologic Neurons Nuclear Pain Pathogenesis Peripheral Postherpetic neuralgia Process Proteins Quality of life Recombinants Refractory Reporter Reporting Research Rodent Role Site Source System Testing Time Tissues Trans-Activators Vaccines Viral Viral Genes Viral Proteins Virion Virus Withdrawal anterograde transport base cell type cellular imaging chromatin modification chronic pain gene function genetic regulatory protein high risk human embryonic stem cell human morbidity improved inhibitor/antagonist innovation painful neuropathy prevent protein expression protein function public health relevance reactivation from latency retrograde transport time use treatment strategy viral RNA

项目摘要

DESCRIPTION (provided by applicant): There exists a great need for the modeling of axonal transport and persistence of the human herpesvirus Varicella zoster Virus in neurons. The sensory neuron is critical to successful VZV pathogenesis as it is the site of a decades-long state of persistence, from which VZV can reactivate to cause the debilitating disease Herpes zoster. VZV remains a major source of human morbidity, even in an age of commercial vaccines, as most adults harbor wild type VZV strains and some 1/5 will suffer zoster ("shingles"). Zoster morbidity includes a subsequent chronic intractable neuropathic pain state that can affect quality of life and is often refractory to any treatment. Even if all eligible persns received the zoster vaccine (which is far from being achieved); the partial effectiveness would still result upwards of half as million zoster cases annually and some 50,000 cases of severe post herpetic neuralgia. We know little of the parameters affecting VZV axonal transport and the latent state, because most animal models and their neurological tissues do not support VZV replication or reactivation. Yet, if axonal transport, latency or the interneuronal spread associated with zoster can be prevented, disease could more easily be controlled. Our overlying hypothesis is that we can explore VZV axonal transport and persistence using an innovative system involving peripheral neurons developed from human embryonic stem cells (hESC). We have partly established this system and shown VZV axonal infection and neuron to neuron spread. The first specific aim is to use hESC derived neurons in microfluidic chambers to examine retrograde and anteriograde transport kinetics of VZV capsids and the association of known VZV tegument regulatory proteins with the transporting capsid, using live cell imaging of fluorescent VZV capsids in axons, which has never been previously described. Specific Aim 2 will test the hypothesis that axonal transport and/or interneuronal spread can be disrupted by specific VZV gene deletions. As yet we know little of the VZV proteins involved in VZV transport. The third aim is to develop the hESC neurons to model VZV persistence, the events of latency, and to attempt to reactivate persistent VZV genomes, which heretofore has arguably never been achieved.

描述（由申请人提供）：对神经元中人类疱疹病毒毒病毒带状疱疹病毒的轴突运输和持久性建模非常需要。感觉神经元对于成功的VZV发病机理至关重要，因为它是数十年来持久状态的部位，VZV可以从中重新激活以引起使人衰弱的疾病疱疹带状疱疹。即使在商业疫苗的时代，VZV仍然是人类发病率的主要来源，因为大多数成年人都有野生型VZV菌株，约有1/5会遭受带状牵化器（“带状疱疹”）。带状疱疹的发病率包括随后的慢性顽固性神经性疼痛状态，可能会影响生活质量，并且通常对任何治疗感到难治。即使所有合格的persns都接受了带状疱疹疫苗（远非实现）；由于每年的带状疱疹病例和大约50,000例严重的疱疹后神经痛病例，部分效率仍将导致一半以上的效率。我们几乎不知道影响VZV轴突运输和潜在状态的参数，因为大多数动物模型及其神经组织不支持VZV复制或重新激活。但是，如果可以防止轴突运输，潜伏期或与带状携带的带状疱疹相关的神经元扩散，则可以更容易控制疾病。我们上覆的假设是，我们可以使用涉及从人类胚胎干细胞（HESC）开发的周围神经元的创新系统探索VZV轴突运输和持久性。我们已经部分建立了该系统，并显示了VZV轴突感染和神经元扩散的神经元。第一个具体目的是在微流体室中使用hESC衍生的神经元来检查VZV衣壳的逆行和前进运输动力学，以及已知的VZV TEGUMEMENT调节蛋白与capsid的已知VZV Tegument调节蛋白的关联，并使用荧光内荧光型VZV CAPSID的活细胞成像的轴突的实用细胞成像（以前从未被描述过。特定的目标2将检验以下假设：轴突转运和/或神经元间扩散可能会被特定的VZV基因缺失破坏。到目前为止，我们对VZV运输的VZV蛋白很少知道。第三个目的是开发hESC神经元来对VZV持久性，延迟事件进行建模，并试图重新激活持续的VZV基因组，而迄今为止，这些基因组可以说从未实现。