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

• 批准号: 8487903
• 负责人: Paul R. Kinchington
• 金额: $ 22.1万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8487903
• 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

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）：非常需要对神经元中人疱疹病毒水痘带状疱疹病毒的轴突运输和持久性进行建模。感觉神经元对于成功的VZV发病机制至关重要，因为它是持续数十年的持续状态的部位，VZV可以从中重新激活以引起使人衰弱的疾病带状疱疹。即使在商业疫苗的时代，VZV仍然是人类发病的主要来源，因为大多数成年人携带野生型VZV毒株，并且大约1/5的人将患有带状疱疹（“带状疱疹”）。带状疱疹发病率包括随后的慢性顽固性神经性疼痛状态，可影响生活质量，通常对任何治疗都难以治愈。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）;部分有效性仍然会导致每年超过50万例带状疱疹病例和约50，000例严重的带状疱疹后神经痛。我们知道很少的参数影响VZV轴突运输和潜伏状态，因为大多数动物模型和他们的神经组织不支持VZV复制或重新激活。然而，如果能够预防带状疱疹相关的轴突运输、潜伏期或神经元间扩散，疾病就更容易控制。我们的假设是，我们可以探索VZV轴突运输和持久性使用一个创新的系统，涉及从人胚胎干细胞（hESC）开发的外周神经元。我们已经部分建立了这个系统，并显示VZV轴突感染和神经元到神经元的传播。第一个具体的目的是使用hESC衍生的神经元在微流体室中检查VZV衣壳的逆行和顺行运输动力学和已知的VZV外皮调节蛋白与运输衣壳的关联，使用轴突中的荧光VZV衣壳的活细胞成像，这在以前从未描述过。具体目标2将测试的假设，轴突运输和/或神经元间的传播可以被破坏特定的VZV基因缺失。然而，我们知道很少的VZV蛋白参与VZV的运输。第三个目标是开发hESC神经元来模拟VZV持续性，潜伏期事件，并尝试重新激活持续性VZV基因组，这在此之前可以说从未实现过。