Modeling drugs of abuse-HIV interactions using iPSC-derived human cerebral organoids

使用 iPSC 衍生的人脑类器官模拟药物滥用与 HIV 相互作用

• 批准号: 10491704
• 负责人: Vez REPUNTE-CANONIGO
• 金额: $ 87.11万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10491704
• 关键词: AIDS/HIV problem; Alcohol abuse; Alzheimer' s Disease; Anti-Retroviral Agents; Astrocytes; Automobile Driving; CRISPR/Cas technology; Cells; Cerebrum; Closure by clamp; Computing Methodologies; Data; Disease; Disease Progression; Dissection; Drug Modelings; Drug abuse; Electrophysiology (science); Exposure to; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genomic approach; HIV; HIV Envelope Protein gp120; Human; Inflammation; Injury; Integration Host Factors; Lead; Light; Literature; Mediator of activation protein; Methamphetamine; Microglia; Modeling; Monitor; Morphine; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Neuropathogenesis; Neuropsychology; Opioid; Organoids; Pathogenesis; Persons; Pharmaceutical Preparations; Preparation; Property; Proviruses; Regulation; Regulator Genes; Role; Stimulant; Substance abuse problem; Systems Biology; Testing; Therapeutic; Tissues; Ursidae Family; Viral; Viral Proteins; Viral reservoir; Virus; advanced system; antiretroviral therapy; chemokine; clinically relevant; comorbidity; cytokine; dopaminergic neuron; drug of abuse; druggable target; experience; experimental study; gene regulatory network; genetic manipulation; improved; in vivo; induced pluripotent stem cell; innovation; macrophage; member; multidrug abuse; neural circuit; neuroAIDS; neuroadaptation; neuron loss; neuroprotection; novel; novel therapeutics; promoter; public health relevance; reactivation from latency; response; therapeutic target; therapeutically effective; transcriptomics

Summary HIV-infected microglia release cytokines, chemokines and viral gene products, such as gp120 and tat, that are toxic to neurons. In this proposal, we will use human induced pluripotent stem cells (iPSCs) to generate cortical and dopaminergic neurons as well as microglia, which will be cultured as organoids to model HIV-drug abuse interactions in conjunction with in-depth genomic approaches and electrophysiology. We show electrophysiological results that demonstrate that the levels of differentiation of the neurons in our organoids are comparable to in vivo and ex vivo preparations and are among the best that can be found in the scientific literature. To identify candidate host regulators of HIV expression and mediators of HIV-induced tissue damage and disease progression, we will expose microglia-containing cerebral organoids to methamphetamine (METH), a stimulant, and morphine, an opiate, which are members of two of the classes of drugs of abuse that are more prevalent among People Living with HIV/AIDS (PLWHA). Cerebral organoids will be exposed to toxic HIV products such as Tat, gp120, drugs of abuse and combination antiretroviral therapy (cART). In some experiments we will also incorporate iPSC-derived astrocytes into microglia-containing cerebral organoids to test the role of astrocytes in neuroprotection and neurodegeneration. We will carry out single-cell gene expression profiling of iPSC-derived organoids exposed to drugs of abuse, HIV products, and cART and we will employ an advanced systems biology strategy to generate testable mechanistic hypotheses on the pathogenesis of neurodegeneration and tissue damage in neuroHIV (Aim 1) and identify candidate regulators of the LTR promoter that may shed light on the regulation of latency and reactivation of the HIV provirus (Aim 2). In preliminary studies, this computational experimental approach allowed us to identify candidate drivers of gene expression changes associated with neuroHIV and neurodegenerative diseases, including Alzheimer’s disease and drug and alcohol abuse. These findings support the overarching hypothesis that dissection of the gene regulatory network of the central nervous system will pave the way for the identification of novel mechanistic hypotheses and druggable targets to improve neuropsychological functioning of PLWHA and substance abuse comorbidity.

概括 HIV感染的小胶质细胞释放细胞因子，趋化因子和病毒基因产物，例如GP120和TAT， 对神经元有毒。在此提案中，我们将使用人类诱导的多能干细胞（IPSC）生成 皮质和多巴胺能神经元以及小胶质细胞，将其培养为类和类器官，以模拟HIV-grug 滥用相互作用与深入的基因组方法和电生理学结合。我们显示 电生理结果表明我们类型器中神经元的分化水平 与体内和离体制剂相媲美，并且是科学中可以找到的最好的 文学。确定艾滋病毒表达的候选宿主调节剂和艾滋病毒诱导的组织损伤的介体 和疾病进展，我们将使含小胶质细胞的大脑器官暴露于甲基苯丙胺 （甲基苯丙胺），一种兴奋剂和吗啡，一种优化，是两种滥用药物的成员 在艾滋病毒/艾滋病（PLWHA）的人中更加普遍。大脑器官将暴露于有毒 HIV产品，例如TAT，GP120，滥用和组合抗逆转录病毒疗法（CART）。在某些人中 实验我们还将将IPSC衍生的星形胶质细胞纳入含小胶质细胞的大脑器官 测试星形胶质细胞在神经保护和神经变性中的作用。我们将执行单细胞基因 暴露于滥用药物，艾滋病毒产品和购物车的IPSC衍生的类器官的表达分析，我们 将采用先进的系统生物学策略来生成可检验的机械假设 神经变性和神经性组织损伤的发病机理（AIM 1）并鉴定候选调节剂 LTR启动子可能会阐明HIV病毒的潜伏期和重新激活的调节（目标 2）。在初步研究中，这种计算实验方法使我们能够确定 基因表达与神经hiv和神经退行性疾病有关的变化，包括阿尔茨海默氏症 疾病，药物和酗酒。这些发现支持了总体假设 中枢神经系统的基因调节网络将为识别新颖的道路铺平道路 机械假设和可药物靶标，以改善PLWHA的神经心理功能 药物滥用合并症。