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

• 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
• 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

 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(申请人提供)：非常需要了解人类疱疹病毒水痘带状疱疹病毒(VZV)如何与神经元相互作用。感觉神经元是带状疱疹病毒持续状态持续数十年的场所，也是带状疱疹病毒重新激活的来源，对VZV的成功发病至关重要。三分之一的成年人患有这种令人痛苦和虚弱的疾病，并经常伴有慢性疼痛和神经问题。此外，眼部带状疱疹是一种潜在的破坏性传染病，会导致严重的视力丧失，在疼痛、并发症和生活质量明显中断方面比其他地方的带状疱疹造成更大的损失。即使所有符合条件的人都接种了带状疱疹疫苗(这还远远没有实现)。部分效果仍然会产生 每年约有50万带状疱疹病例。由于大多数动物模型或其神经组织不支持VZV复制或再激活，我们对VZV潜伏期和再激活的了解仍然是谜、有争议和不清楚的。事实上，事实证明，在任何情况下，VZV的实验重新激活都是极其困难的。然而，如果能够防止神经元感染、扩散、持续或重新激活，疾病可能会更容易控制。我们现在有了一个前所未有的培养的人类神经元平台，来自人类胚胎干细胞，不仅支持VZV生产性感染，而且支持现在可以重新激活的持续状态。我们的假设是，这个系统将使我们能够以一种以前不可能的方式解决神经元感染、潜伏期和Q重新激活的各个方面。我们的第一个目标是验证特定的VZV蛋白是必需的假设。 对于神经元轴突感染、复制、神经元间扩散或顺行轴突返回。我们将评估荧光报告VZV与神经元感染每个阶段的基因缺失，以确定所需的蛋白质。这样的蛋白质不仅可以作为阻断潜伏期和重新激活的策略的靶点，而且这种VZV突变可能成为具有明确的神经趋向性缺陷的改进疫苗的基础。我们的第二个目标将解决VZV持续和重新激活期间的病毒转录程序。然后，我们将确定在持续和重新激活期间发生了什么转录，包括寻找可能导致延迟的小RNA。我们还将解决在不同温度下重新激活后转录程序是否有所不同。我们的第三个目标将检验这一假设，即持久的VZV基因组可以被大序列识别核酸酶靶向，以减少VZV潜在的基因组负荷或防止重新激活。这将确立原则或针对潜伏的VZV基因组，而不依赖病毒基因产品。在全球范围内，我们的研究将为理解 并且以以前不可能的方式瞄准VZV潜伏期。