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

描述(由申请人提供)：非常需要对人类疱疹病毒水痘带状疱疹病毒在神经元中的轴突运输和持久性进行建模。感觉神经元是VZV成功发病的关键，因为它是长达数十年的持续状态的场所，VZV可以从这个状态重新激活，导致衰弱的带状疱疹。即使在商业疫苗时代，VZV仍然是人类发病率的主要来源，因为大多数成年人携带野生型VZV毒株，约五分之一的人将患带状疱疹(带状疱疹)。带状疱疹的发病率包括随后的慢性顽固性神经病理性疼痛状态，这种疼痛状态会影响生活质量，而且通常对任何治疗都难以奏效。即使所有符合条件的人都接种了带状疱疹疫苗(这还远未实现)，部分疗效仍将导致每年50万带状疱疹病例和约50,000例严重带状疱疹后神经痛。我们对影响VZV轴突运输和潜伏状态的参数知之甚少，因为大多数动物模型及其神经组织不支持VZV的复制或再激活。然而，如果与带状疱疹相关的轴突运输、潜伏期或神经元间扩散能够被阻止，疾病就可以更容易地得到控制。我们的假设是，我们可以使用一种创新的系统来探索VZV的轴突运输和持久性，该系统涉及到从人类胚胎干细胞(HESC)发展而来的外周神经元。我们已经部分建立了这个系统，并显示了VZV轴突感染和神经元到神经元的扩散。第一个特定的目的是利用微流室中的hESC来源的神经元来研究VZV衣壳的逆行和顺行运输动力学，以及已知的VZV被膜调节蛋白与运输衣壳的联系，使用以前从未描述过的轴突中的VZV荧光衣壳的活细胞成像。《特定目标2》将验证一种假说，即特定的VZV基因缺失可以破坏轴突运输和/或神经元间扩散。到目前为止，我们对参与VZV转运的VZV蛋白知之甚少。第三个目标是发展hESC神经元来模拟VZV持续潜伏期事件，并试图重新激活VZV持续基因组，这是迄今为止可以说从未实现过的。

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.