Effects of HSV-1 infection on neural progenitor cell biology in vitro and in vivo

8.摘要 我们将研究单纯疱疹病毒1型（HSV-1）感染神经祖细胞（NPC） 在R 01应用程序中。在免疫功能正常的个体中，HSV-1感染是最常见的 脑炎（HSE）虽然抗病毒治疗改善了其预后，但大多数 的HSV-1脑炎幸存者患有永久性神经系统后遗症。慢性病变 在HSE患者中观察到与记忆形成相关的区域，其中包括 海马体和相关的边缘结构。HSE后遗症的严重程度与 对这些地区的破坏程度。 HSV-1表现出对海马（颗粒下区，SGZ）和脑室下区的嗜性 (SVZ)侧脑室这两个区域都富含NPC，它们分化成神经元， 神经胶质细胞因此，这些区域是成人神经发生的主要生态位区域。尽管关键的 NPC在成体神经发生中的作用，HSV-1对NPC增殖的影响， 分化和迁移在很大程度上未知。此外，NPC是否可以代表 CNS中HSV-1的储库是未知的。 在这里，我们的目标是使用三维（3D）培养来模拟HSV-1与NPC的相互作用 从诱导多能干细胞（hiPSC）衍生的人类CNS细胞;我们还将建立在 小鼠脑炎模型，使实验无法在体外人类细胞。 在目的1中，我们将研究HSV-1感染对增殖、分化 以及在三维（3D）培养系统中源自hiPSC的NPC的迁移， 设计;该模型包含NPC及其衍生物。在目标2中，我们将采用 小鼠脑炎模型以研究HSV-1在海马和SVZ中的分布， HSV-1感染对NPC增殖的影响，并分析存活小鼠中的NPC， 期限为6个月至1年。此外，我们将确定是否有HSV-1的证据 NPC的潜伏和复活。在目标3中，我们将研究：i）NPC中的HSV-1潜伏期; ii）NPC中的HSV-1潜伏期。 HSV-1感染对调节NPC分化命运途径的影响 调节;和iii）持续感染对NPC功能的后果。啮齿动物和 将对iPSC数据进行合成。我们假设i）HSV-1损害NPC过程所需的 ii）HSV-1可以在NPC中建立潜伏期。 从这个项目中所学到的将提高我们对HSV-1脑炎的认识。我们 具备拟开展工作的相关专业知识。

8. Abstract We will investigate Herpes simplex virus type 1 (HSV-1) infection of neural progenitor cells (NPCs) in our R01 application. In immunocompetent individuals, HSV-1 infection is the most common cause of encephalitis (HSE). Although antiviral therapy has improved its prognosis, the majority of HSV-1 encephalitis survivors suffer from permanent neurological sequelae. The chronic lesions in HSE patients have been observed in regions associated with memory formation, which include the hippocampus and associated limbic structures. The severity of sequela in HSE is related to the severity of damage to these regions. HSV-1 displays tropism for hippocampus (subgranular zone, SGZ) and the subventricular zone (SVZ) of the lateral ventricles. Both regions are rich in NPCs, which differentiate into neurons and glial cells. These regions are thus major niche areas for adult neurogenesis. Despite the pivotal role played by the NPCs in adult neurogenesis, the effect of HSV-1 on NPCs proliferation, differentiation and migration is largely unknown. Furthermore, whether NPCs may represent reservoirs of HSV-1 in the CNS is unknown. Here, we aim to model the interaction of HSV-1 with NPCs using three-dimensional (3D) cultures of human CNS cells derived from induced pluripotent stem cells (hiPSCs); we will also build on a mouse model of encephalitis to enable experiments not feasible in human cells in vitro. In Aim 1, we will investigate early-stage effects of HSV-1 infection on proliferation, differentiation, and migration of NPCs derived from hiPSCs in three-dimensional (3D) culture systems that we have designed; the model incorporates NPCs and their derivatives. In Aim 2, we will employ a mouse model of encephalitis to investigate the distribution of HSV-1 in the hippocampus and SVZ, the effects of HSV-1 infection on NPCs proliferation and analyze NPCs in surviving mice for periods ranging from 6 months to 1 year. Also, we will determine if there is evidence of HSV-1 latency and reactivation in NPCs. In Aim 3, we will investigate: i) HSV-1 latency in NPCs; ii) the effect of HSV-1 infection on pathways regulating the differentiation fate of NPCs pathways regulating; and iii) the consequences of persistent infection on NPCs function. The rodent and the iPSC data will be synthesized. We hypothesize that i) HSV-1 impairs NPC processes required for neurogenesis; ii) HSV-1 can establish latency in NPCs. What is learned from this project will improve our understanding of HSV-1 encephalitis. We possess the relevant expertise for the proposed work.