Modeling HIV-associated neurocognitive disorders and encephalopathy in human iPSC brain organoids containing microglia

在含有小胶质细胞的人 iPSC 脑类器官中模拟 HIV 相关的神经认知障碍和脑病

• 批准号: 10266193
• 负责人: RONALD P HART
• 金额: $ 23.55万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10266193
• 关键词: 3-Dimensional; Address; Adult; Affect; Animal Model; Anti-HIV Therapy; Behavioral; Biological Models; Brain; Cell Communication; Cell Lineage; Cell model; Cells; Cerebrum; Child; Clinical; Cognition Disorders; Cognitive; Cultured Cells; Defect; Development; Disease; Encephalopathies; Functional disorder; Gene Expression; Generations; Goals; HIV; HIV Encephalopathy; HIV Infections; HIV-associated neurocognitive disorder; Human; Immune; Infection; Inflammatory; Lead; Mediator of activation protein; Microglia; Modeling; Molecular; Molecular Analysis; Monitor; Motor; Neurodevelopmental Deficit; Neuroimmune; Neurons; Neurosciences; Nuclear Pore Complex; Organoids; Pathogenesis; Pathogenicity; Patients; Phenotype; Population; Positioning Attribute; Production; Public Health; Role; Severities; Symptoms; System; Testing; Tissues; Virus Diseases; antiretroviral therapy; base; brain cell; cell assembly; clinical phenotype; cytokine; experimental study; induced pluripotent stem cell; innovation; insight; multidisciplinary; nerve stem cell; nervous system disorder; neurodevelopment; neurophysiology; neuropsychiatric disorder; novel; novel therapeutics; pediatric human immunodeficiency virus; relating to nervous system; single-cell RNA sequencing; stem cell biology; stem cell technology; tool; transcriptomics; two-dimensional; virology

Project Summary/Abstract Even with the advent of successful combination antiretroviral therapy (c-ART), infection by Human Immunodeficiency Virus (HIV-1) remains a global public health crisis. HIV infection of the brain results in important clinical symptoms including HIV-associated neurocognitive disorder or HAND, a constellation of symptoms affecting cognitive, behavioral, and motor functions, as well as pediatric HIV encephalopathy (PHE) in children, leading to neurodevelopmental deficits. These disorders may still occur even in patients receiving c- ART, although with less severity. The molecular and cellular mechanisms underlying the pathophysiology of HIV- HAND and PHE remain poorly understood, in large part because of limitations of the existing model systems to study them. Animal models are generally not infected by HIV, expensive and impractical or poorly recapitulate human brain infection. Cultured cell models provide some insights, but do not recapitulate many aspects of cell- cell and cell-matrix interactions found in human brain. Advances in stem cell technologies, including the use of human induced pluripotent stem cells (iPSC or iPS cells), now enable generation of patient-specific neural lineage cells and microglia (brain immune cells), as well as the assembly of these cells into 3-dimensional (3D) brain organoids. These cerebral organoids mimic important features of immune, glial and neuronal cell interactions, uniquely enabling us to examine how these interactions are affected by HIV infection and to model key aspects of HAND and PHE pathogenesis. The overarching goal of this project is to leverage our strengths in neurosciences, virology, stem cell biology and gene expression to understand the pathophysiology of HAND and PHE using a human neural-microglial system. We have established human 3D brain organoids prepared with iPS cell-derived microglial cells to study neurodevelopment and neuroimmune interactions and have successfully infected these mixed organoids with HIV. We hypothesize that microglial infection by HIV causes altered neuronal function which can be modeled in these microglial-containing brain organoids, providing a new system for mechanistic understanding of the pathogenesis of HAND and PHE. In this proposal, we will characterize the effects of HIV infection on microglia activation and cytokine production, neuronal cell populations, organization and gene expression, and the specific effects of candidate cytokine mediators. We will apply cutting- edge approaches drawn from stem cell technologies, brain organoid models, and single cell RNA sequencing to develop this model system, which will allow long-term, detailed analyses of the molecular and neurophysiologic mechanisms responsible for the pathogenesis of HIV-associated neurological disorders.

项目概要/摘要 即使出现了成功的联合抗逆转录病毒疗法（c-ART），人类感染 免疫缺陷病毒（HIV-1）仍然是全球公共卫生危机。大脑感染艾滋病毒会导致 重要的临床症状，包括 HIV 相关的神经认知障碍或 HAND（一系列症状） 影响认知、行为和运动功能以及儿童艾滋病毒脑病 (PHE) 的症状 对儿童来说，会导致神经发育缺陷。即使在接受 c-治疗的患者中，这些疾病仍然可能发生 ART，尽管严重程度较低。 HIV病理生理学的分子和细胞机制 HAND 和 PHE 仍然知之甚少，很大程度上是因为现有模型系统的局限性 研究他们。动物模型一般不会感染艾滋病毒，价格昂贵且不切实际或重现性差 人脑感染。培养细胞模型提供了一些见解，但没有概括细胞的许多方面 人脑中发现的细胞和细胞-基质相互作用。干细胞技术的进步，包括使用 人类诱导多能干细胞（iPSC 或 iPS 细胞）现在能够生成患者特异性神经细胞 谱系细胞和小胶质细胞（大脑免疫细胞），以及将这些细胞组装成 3 维 (3D) 脑类器官。这些大脑类器官模仿免疫细胞、神经胶质细胞和神经元细胞的重要特征 相互作用，使我们能够独特地研究这些相互作用如何受到 HIV 感染的影响并建立模型 HAND 和 PHE 发病机制的关键方面。该项目的总体目标是发挥我们的优势 神经科学、病毒学、干细胞生物学和基因表达领域的研究人员，以了解 HAND 的病理生理学 以及使用人类神经小胶质细胞系统的 PHE。我们已经建立了人类 3D 大脑类器官 与 iPS 细胞衍生的小胶质细胞一起研究神经发育和神经免疫相互作用，并具有 成功地用艾滋病毒感染了这些混合类器官。我们假设 HIV 感染小胶质细胞会导致 神经元功能的改变可以在这些含有小胶质细胞的大脑类器官中进行建模，从而提供了一种新的方法 系统了解 HAND 和 PHE 的发病机制。在本提案中，我们将 描述 HIV 感染对小胶质细胞激活和细胞因子产生、神经元细胞群的影响， 组织和基因表达，以及候选细胞因子介质的具体作用。我们将应用切割- 来自干细胞技术、脑类器官模型和单细胞 RNA 测序的前沿方法 开发这个模型系统，这将允许对分子和神经生理学进行长期、详细的分析 HIV 相关神经系统疾病的发病机制。