Molecular studies of VZV infection, latency and reactivation in human neurons in-vitro

人类神经元水痘带状疱疹病毒感染、潜伏期和再激活的分子研究

基本信息

批准号：
9179591
负责人：
Paul R. Kinchington
金额：
$ 45.79万
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：
2015
资助国家：
美国
起止时间：
2015-11-12 至 2020-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9179591
关键词：
Address Adult Afferent Neurons Age Animal Model Axon Blindness Capsid Chickenpox Climacteric Collaborations Communicable Diseases Defect Disease Effectiveness Eligibility Determination Event Eye diseases Foundations Gene Deletion Gene Expression Genes Genetic Transcription Genomics Growth Herpes zoster disease Herpesviridae Herpesvirus Type 3 Homing Human Immune In Vitro Individual Infection Knowledge Latent Virus Libraries Life Light Lytic Modeling Molecular Nerve Neurologic Neurons Neurotropism Outcome Pain Peripheral Persons Prevention strategy Process Proteins Quality of life Recombinants Reporter Research Personnel Risk Role Site Small RNA Source Stimulus System Temperature Testing Therapeutic Tissues Vaccines Viral Viral Genes Viral Genome Viral Proteins Virus Virus Diseases Virus Replication Work Zoster Vaccine chronic pain design experimental study genetic regulatory protein human embryonic stem cell improved mutant neuronal cell body next generation sequencing novel nuclease prevent programs prophylactic public health relevance vector viral RNA virus genetics virus pathogenesis

项目摘要

DESCRIPTION (provided by applicant): There exists a great need to understand how the human herpesvirus Varicella zoster Virus (VZV) interacts with neurons. The sensory neuron is critical to successful VZV pathogenesis as the site of a decades-long state of persistence, and the source for VZV reactivation to cause Herpes zoster. This painful and debilitating disease is encountered by a third of adults, and is frequently complicated by chronic pain and neurological problems. Furthermore, ophthalmic zoster is a potentially devastating infectious disease that causes significant vision loss and a much greater toll than zoster elsewhere in terms of pain, complications and marked disruption of the quality of life. Even if all eligible persons received the zoster vaccine (which is far from being achieved). the partial effectiveness would still result in some half of a million zoster cases annually. What we know of VZV latency and reactivation has remained enigmatic, controversial and unclear, because most animal models or their neurological tissues do not support VZV replication or reactivation. Indeed, VZV experimental reactivation has proved exceedingly difficult under any circumstances. Yet, if neuronal infection, spread, persistence or reactivation can be prevented, disease could be more easily controlled. We now have an unprecedented cultured human neuron platform, derived from human embryonic stem cells, that supports not only VZV productive infection, but also persistent states that can now be reactivated. Our overlying hypothesis is that this system will enable us to address aspects of neuron infection, latency and qreactivation in a manner that has not been previously possible. Our first aim will test the hypothesis that specific VZV proteins are required for neuron axonal infection, replication, inter-neuronal spread or anterograde axonal return. We will evaluate fluorescent reporter VZV with gene deletions for each stage of neuronal infection to identify those proteins required. Such proteins could be not only be targeted in strategies to block latency and reactivation, but such VZV mutants could be the basis for improved vaccines with defined deficiencies in neurotropism. Our second aim will address the viral transcription program during VZV persistence and reactivation. We will then identify what transcription occurs during persistence and reactivation, including a search for small RNAs that may contribute to latency. We will also address if transcriptional programs differ following reactivation at differen temperatures. Our third aim will test the hypothesis that the persistent VZV genome can be targeted by large sequence recognition nucleases, in order to reduce VZV latent genomic loads or prevent reactivation. This would establish principles or targeting the latent VZV genome without reliance on viral gene products. Globally, our studies will set the stage for understanding and targeting the VZV latent state in a manner that has not been previously possible.

描述（由适用提供）：非常需要了解人类疱疹病毒带状疱疹病毒（VZV）如何与神经元相互作用。感觉神经元对于成功的VZV发病机理至关重要，作为数十年的持久状态的部位，也是VZV重新激活引起带状疱疹的来源。这种痛苦和使人衰弱的疾病遇到了三分之一的成年人，并且经常因慢性疼痛和神经系统问题而复杂化。此外，在疼痛，并发症和生命质量上的明显中断方面，眼科系绳是一种潜在毁灭性的意外疾病，它会造成大量视力丧失，并且比其他地方更大。即使所有合格的人都接受了带状疱疹疫苗（远非实现）。部分有效性仍然会导致在每年的带领案例中，大约一半。我们对VZV潜伏期和重新激活的了解一直存在着神秘的，有争议的和不清楚的，因为大多数动物模型或其神经组织不支持VZV复制或重新激活。实际上，在任何情况下，VZV实验重新激活都更加困难。但是，如果可以预防神经元感染，扩散，持久性或重新激活，则可以更容易控制疾病。现在，我们拥有一个前所未有的培养的人类神经元平台，该平台源自人类胚胎干细胞，不仅支持VZV生产感染，而且支持现在可以重新激活的持久状态。我们上覆的假设是，该系统将使我们能够以以前不可能的方式解决神经元感染，潜伏期和QREACTIVATION的各个方面。我们的第一个目标将检验需要特定VZV蛋白的假设用于神经元感染，复制，神经间扩散或顺行轴突回流。我们将在神经元感染的每个阶段用基因缺失评估荧光记者VZV，以鉴定所需的蛋白质。这种蛋白质不仅可以针对阻断潜伏期和重新激活的策略，而且可以成为改善神经营养缺乏症的疫苗的基础。我们的第二个目标将解决VZV持久性和重新激活期间的病毒转录程序。然后，我们将确定在持久性和重新激活期间发生的转录，包括搜索可能导致潜伏期的小型RNA。在不同温度下重新激活后，我们还将解决转录程序是否有所不同。我们的第三个目标将检验以下假设：持续的VZV基因组可以由大序列识别核者靶向，以减少VZV潜在的基因组载荷或防止重新激活。这将建立原理或靶向潜在的VZV基因组，而不依赖病毒基因产物。在全球范围内，我们的研究将为理解奠定基础并以先前无法实现的方式瞄准VZV潜在状态。