Cerebral organoid and IPSC derived microglia: Modeling of HIV and methamphetamine co-morbidity

大脑类器官和 IPSC 衍生的小胶质细胞：HIV 和甲基苯丙胺共病模型

• 批准号: 10687067
• 负责人: Peng Jiang
• 金额: $ 53.11万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10687067
• 关键词: 3-Dimensional; Abuse Reporting; Acquired Immunodeficiency Syndrome; Address; Adolescent; Adult; Anti-Inflammatory Agents; Anti-Retroviral Agents; Applications Grants; Astrocytes; Autopsy; Biogenesis; Biological Assay; Biology; Blood - brain barrier anatomy; Brain; CRISPR/Cas technology; Cells; Central Nervous System; Cerebrum; Chronic; Clinical; Coculture Techniques; Data; Dendritic Spines; Development; Diagnosis; Disease; Drug abuse; End Point Assay; Ensure; Erythro; Etiology; Event; Genes; Goals; HIV; HIV Genome; HIV Infections; HIV-associated neurocognitive disorder; High Prevalence; Human; Individual; Inflammation; Inflammatory; Knowledge; Macrophage; Measures; Mediating; Methamphetamine; MicroRNAs; Microglia; Mitochondria; Modeling; Molecular; Molecular Profiling; Monkeys; Mus; Myelogenous; Nerve Degeneration; Neuroglia; Neuroimmune; Neuronal Injury; Neurons; Neuropathogenesis; Organoids; Outcome; Pathogenesis; Pathology; Pathway interactions; Patients; Pattern; Penetration; Persons; Pharmaceutical Preparations; Population; Property; Proteins; Regimen; Reproducibility; Role; SIV; Sampling; Series; Signal Pathway; Signal Transduction; Societies; Synapses; Technology; Toxic effect; Treatment Efficacy; Viral; Virus Diseases; Yolk Sac; antiretroviral therapy; brain dysfunction; brain tissue; cell type; comorbidity; design; drug of abuse; experimental study; extracellular vesicles; genome editing; graft vs host disease; human pluripotent stem cell; humanized mouse; in vivo; induced pluripotent stem cell; metabolic rate; methamphetamine abuse; neuroinflammation; neurotoxicity; non-invasive imaging; novel; novel therapeutics; pharmacokinetics and pharmacodynamics; postsynaptic; presynaptic; progenitor; progenitor system; psychostimulant; receptor; response; self organization; single-cell RNA sequencing; success; synaptic pruning; therapeutic evaluation; three-dimensional modeling; tool

Abstract: An estimated ~1.3 million adults and adolescents are living with human immunodeficiency virus (HIV) infection in the USA of which 15% of them undiagnosed. The initiation of combination antiretroviral therapy (cART) has transformed AIDS from a fatal illness into a chronic and manageable disease. The presence of HIV infected microglia and macrophage reservoirs in the central nervous system (CNS) and variable penetration of anti- retroviral drugs across the blood brain barrier after cART are likely factors for the persistence of HIV associated neurocognitive disorder (HAND). Drugs of abuse such as the potent psychostimulant methamphetamine (Meth) that is abused by an estimated 30 million people in the world further minimize the efficacy of cART and in conjunction with HIV exacerbate CNS pathology. Thus, significant gap in knowledge in the field is to better understand the neuropathogenesis and the etiology of clinical outcomes observed in HIV+ patient’s dependent on Meth abuse. The current grant proposal focuses on developing a human induced pluripotent stem cell based cerebral organoid model to investigate underlying brain dysfunction during HIV/Meth interaction. Cerebral organoids are 3-dimensional “mini brains” that can self-organize and recapitulate many milestone events seen in in vivo brain development. Our recent study on generating a novel organoid model with the feasibility in controlling the microglia ratio and microenvironment of organoid formation to recapitulate changes in brain functions under various conditions (e.g. virus and viral infection-induced inflammation and synaptic pruning) makes this model ideal for our proposed studies. The central hypothesis of this proposal is that Meth in conjunction with HIV causes significant neuronal damage. The combination of microglia-containing cerebral organoid model, extracellular vesicle biology, single-cell RNA-sequencing, CRISPR/Cas9-mediated gene editing will decipher novel intercellular and molecular mechanisms and pathways that underlie abnormalities in neuronal functions and connectivity caused by HIV infection. We will investigate this hypothesis under three specific aims: Specific Aim 1 will examine neuronal function, synaptic and mitochondrial perturbations during HIV/meth interactions in 3D microglia-containing cerebral organoid model; Specific Aim 2 will investigate the role of extracellular vesicles in microglial organoids treated under HIV/meth conditions; and Specific Aim 3 will characterize the molecular mechanisms underlying HIV infection-induced neuronal injury and further evaluate our microglial organoid model as a reliable tool to identify molecular signatures of HAND. Our proposed experiments will decipher molecular mechanisms, novel signaling events and molecular partners underlying the neuronal injury in HAND. Through the series of the experiments we aim to present an easily scalable and reproducible model to study HAND pathogenesis. The data obtained from these studies can be used to design novel therapeutics to control HIV infection.

摘要： 据估计，约有130万成年人和青少年感染了人类免疫缺陷病毒(HIV) 在美国，其中15%的人没有得到诊断。开始联合抗逆转录病毒治疗(CART) 将艾滋病从一种致命的疾病转变为一种慢性和可控的疾病。艾滋病病毒感染者的存在 中枢神经系统(CNS)中的小胶质细胞和巨噬细胞储存库与抗-HBs的可变穿透 逆转录病毒药物在CART后跨越血脑屏障可能是HIV相关病毒持续存在的因素 神经认知障碍(手)。滥用药物，如强效精神刺激剂甲基苯丙胺(Meth) 据估计，世界上有3000万人滥用这种药物，从而进一步降低了Cart和In的功效 与艾滋病毒一起加重了中枢神经系统的病理。因此，在该领域的知识差距显著是为了更好地 了解HIV+患者依赖的神经发病机制和临床结局的病因 关于滥用毒品的指控。目前的拨款方案重点是开发一种基于人类诱导的多能干细胞 研究HIV/Meth相互作用过程中潜在脑功能障碍的脑器官模型。大脑 有机体是一种三维的“迷你大脑”，可以自我组织和概括许多重大事件。 在活体大脑发育中。我们最近关于生成一种新的有机物模型的研究，该模型在 控制小胶质细胞比率和类器官形成的微环境以概括脑内的变化 在不同条件下的功能(例如病毒和病毒感染引起的炎症和突触修剪) 这使得这个模型非常适合我们提议的研究。这一提议的中心假设是，梅斯在 与艾滋病毒结合会造成严重的神经元损伤。含小胶质细胞的脑联合移植 类有机物模型、胞外囊泡生物学、单细胞RNA测序、CRISPR/Cas9介导的基因编辑 将破译神经元异常背后的新的细胞间和分子机制和途径 由艾滋病毒感染引起的功能和连接。我们将在三个具体目标下调查这一假设： 特定目标1将检查HIV/Meth过程中的神经元功能、突触和线粒体的扰动 含小胶质细胞的3D脑器官模型中的相互作用；特异性目标2将研究 在艾滋病毒/冰毒条件下治疗的小胶质细胞有机体中的细胞外小泡；以及特定目标3将 描述HIV感染导致神经元损伤的分子机制，并进一步评估 我们的小胶质细胞器模型是识别手部分子特征的可靠工具。我们的建议 实验将破译分子机制、新的信号事件和潜在的分子伙伴 手部神经元损伤。通过一系列的实验，我们的目标是呈现一种易于扩展和 研究手部发病机制的重复性模型。从这些研究中获得的数据可以用来设计 控制艾滋病毒感染的新疗法。