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: 10623148
• 负责人: David C. Bloom
• 金额: $ 37.16万
• 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/10623148
• 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; Gene Expression; Genes; Genome; Goals; Herpes Simplex Infections; Herpes encephalitis; Herpesvirus 1; Hippocampus; Human; Immunocompetent; Impairment; In Vitro; Incidence; Individual; Infection; Investigation; Language; Learning; Lesion; Limbic System; Lytic; Memory; Modeling; Mus; Neonatal; Neuroglia; Neurologic; Neurologic Deficit; Neurons; Neurosphere; Organoids; Pathogenesis; Pathway interactions; Patients; Periodicals; Phase; Play; Prognosis; Proliferating; Protocols documentation; Resolution; Rodent; Role; Running; Severities; Simplexvirus; Structure; Survivors; System; Technology; Tropism; Virus; Work; adult neurogenesis; aged; 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; permissiveness; prevent; progenitor; programs; reactivation from latency; stem cell function; stem cell model; stem cell proliferation; subventricular zone; three-dimensional modeling; transcriptome; transcriptome sequencing; vaccine access

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 1 型单纯疱疹病毒 (HSV-1) 感染神经祖细胞 (NPC) 的情况 在我们的 R01 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）的原因。尽管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 这些地区受损的严重程度。 HSV-1 显示海马体（颗粒下区，SGZ）和室下区的趋向性 侧脑室 (SVZ)。这两个区域都富含 NPC，可分化为神经元和 胶质细胞。因此，这些区域是成人神经发生的主要利基区域。尽管至关重要 NPC 在成人神经发生中的作用、HSV-1 对 NPC 增殖的影响、 分化和迁移在很大程度上是未知的。此外，NPC 是否可以代表 HSV-1 在 CNS 中的储存库尚不清楚。 在这里，我们的目标是使用三维 (3D) 培养物模拟 HSV-1 与 NPC 的相互作用 源自诱导多能干细胞 (hiPSC) 的人类中枢神经系统细胞；我们还将在此基础上 小鼠脑炎模型使在体外人类细胞中不可行的实验成为可能。 在目标 1 中，我们将研究 HSV-1 感染对增殖、分化、 以及来自 hiPSC 的 NPC 在三维 (3D) 培养系统中的迁移 已设计；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，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的 NPC 期限从6个月到1年不等。此外，我们将确定是否有 HSV-1 的证据 NPC 中的潜伏期和重新激活。在目标 3 中，我们将研究： i) HSV-1 在 NPC 中的潜伏期； ii) 的 HSV-1感染对调控NPCs分化命运途径的影响 调节； iii) 持续感染对 NPC 功能的影响。啮齿动物和 iPSC 数据将被合成。我们假设 i) HSV-1 损害所需的 NPC 过程 用于神经发生； ii) HSV-1 可以在 NPC 中建立潜伏期。 从这个项目中学到的知识将提高我们对 HSV-1 脑炎的了解。我们 拥有拟议工作的相关专业知识。