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

描述（由申请人提供）：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续存在的建模有很大的需求。感觉神经元对VZV的成功发病至关重要，因为它是VZV长达数十年持续状态的部位，从中VZV可以重新激活，导致使人衰弱的带状疱疹疾病。即使在商业化疫苗的时代，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，大约1/5的人会患带状疱疹（带状疱疹）。带状疱疹发病率包括随后的慢性难治性神经性疼痛状态，可影响生活质量，并且通常对任何治疗都是难治性的。即使所有符合条件的人都接种了带状疱疹疫苗（这远远没有实现）；部分有效性仍然会导致每年50万带状疱疹病例和约5万例严重的带状疱疹后神经痛。我们对影响VZV轴突转运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复制或再激活。然而，如果能预防与带状疱疹相关的轴突转运、潜伏期或神经元间扩散，疾病将更容易控制。我们的首要假设是，我们可以利用一种涉及人类胚胎干细胞（hESC）发育的外周神经元的创新系统来探索VZV轴突转运和持久性。我们已经部分建立了这个系统，并显示了VZV轴突感染和神经元间的传播。第一个具体目标是使用hESC衍生的神经元在微流控室中检测VZV衣壳的逆行和顺行运输动力学以及已知VZV被膜调节蛋白与运输衣壳的关联，使用轴突荧光VZV衣壳的活细胞成像，这在以前从未被描述过。特异性目的2将验证轴突运输和/或神经元间传播可以被特异性VZV基因缺失破坏的假设。到目前为止，我们对参与VZV运输的VZV蛋白知之甚少。第三个目标是开发hESC神经元来模拟VZV持久性，潜伏期事件，并尝试重新激活持久性VZV基因组，迄今为止，这可能从未实现过。

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.