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

HSV-1感染对神经祖细胞体外和体内生物学的影响

• 批准号: 10047416
• 负责人: David C. Bloom
• 金额: $ 45.06万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10047416
• 关键词: 3-Dimensional; Acute; Acyclovir; Animal Model; Antigens; Antiviral Agents; Antiviral Therapy; Area; Biology; Brain; Cell Differentiation process; Cell physiology; Cells; Cellular biology; Central Nervous System Diseases; Central Nervous System Infections; Chronic; Complement; Confocal Microscopy; Data; Dose; Encephalitis; Expression Profiling; Gene Expression; Genes; Genome; Goals; Herpes encephalitis; Herpesvirus 1; Hippocampus (Brain); Human; Immunocompetent; Impairment; In Vitro; Incidence; Individual; Infection; Investigation; Language; Lesion; Limbic System; Lytic; Memory; Modeling; Mus; Neonatal; Neuroglia; Neurologic; Neurologic Deficit; Neurons; Organoids; Pathogenesis; Pathway interactions; Patients; Periodicity; Phase; Play; Protocols documentation; Resolution; Rodent; Role; Running; Severities; Simplexvirus; Structure; Survivors; System; Technology; Tropism; Vaccines; Virus; Work; adult neurogenesis; aged; base; biological systems; cell motility; chronic infection; design; executive function; experimental study; human model; improved; in vitro Model; in vivo; induced pluripotent stem cell; insight; latent infection; lateral ventricle; migration; mortality; mouse model; nerve stem cell; neurogenesis; outcome forecast; prevent; progenitor; programs; stem cell model; stem cell proliferation; subventricular zone; three dimensional cell culture; three-dimensional modeling; transcriptome; transcriptome sequencing

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.

8。摘要 我们将研究神经祖细胞（NPC）的1型单纯疱疹病毒（HSV-1）感染 在我们的R01应用程序中。在免疫能力的个体中，HSV-1感染是最常见的 脑炎的原因（HSE）。尽管抗病毒疗法改善了预后，但大多数 HSV-1脑炎幸存者患有永久神经后遗症。慢性病变 在与记忆形成相关的区域中已经观察到HSE患者，其中包括 海马和相关的边缘结构。 HSE中后遗症的严重程度与 对这些地区的损害严重程度。 HSV-1显示海马（晶体下区域，SGZ）和室室内区域 （SVZ）的侧心室。这两个区域都有丰富的NPC，它们分化为神经元和 神经胶质细胞。因此，这些区域是成人神经发生的主要利基区域。尽管有关键 NPC在成人神经发生中扮演的角色，HSV-1对NPCS增殖的影响， 分化和迁移在很大程度上是未知的。此外，是否可以代表NPC 中枢神经系统中HSV-1的储层是未知的。 在这里，我们旨在使用三维（3D）培养物对HSV-1与NPC的相互作用进行建模 源自诱导多能干细胞（HIPSC）的人类中枢神经系统细胞的；我们还将建立 脑炎的小鼠模型使实验在体外人类细胞中不可行。 在AIM 1中，我们将研究HSV-1感染对增殖，分化， 以及在我们的三维（3D）培养系统中源自HIPSC的NPC的迁移 已经设计了；该模型结合了NPC及其衍生物。在AIM 2中，我们将采用 脑炎的小鼠模型研究HSV-1在海马和SVZ中的分布， HSV-1感染对NPCS增殖的影响并分析幸存小鼠中的NPC的影响 期限从6个月到1年不等。另外，我们将确定是否有HSV-1的证据 NPC中的潜伏期和重新激活。在AIM 3中，我们将调查：i）NPC中的HSV-1潜伏期； ii） HSV-1感染对调节NPCS途径分化命运的途径的影响 调节； iii）持续感染对NPC功能的后果。啮齿动物和 IPSC数据将合成。我们假设I）HSV-1会损害所需的NPC过程 用于神经发生； ii）HSV-1可以在NPC中建立延迟。 从这个项目中学到的将提高我们对HSV-1脑炎的理解。我们 拥有拟议工作的相关专业知识。