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.

总结 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，如gp 120和达特， 对神经元有毒在这项提案中，我们将使用人类诱导多能干细胞（iPSC）来产生 皮质和多巴胺能神经元以及小胶质细胞，它们将被培养为类器官以模拟HIV药物 滥用相互作用与深入的基因组方法和电生理学相结合。我们表明 电生理结果表明我们类器官中神经元的分化水平 与体内和离体制剂相当，并且是在科学研究中可以找到的最好的制剂之一。 文学鉴定HIV表达的候选宿主调节因子和HIV诱导的组织损伤介质 和疾病进展，我们将暴露含有小胶质细胞的脑类器官甲基苯丙胺 （METH），一种兴奋剂，和吗啡，一种阿片类药物，这是两类滥用药物的成员， 在艾滋病毒/艾滋病感染者（PLWHA）中更为普遍。大脑类器官将暴露于有毒物质 艾滋病毒产品，如达特、gp 120、滥用药物和联合抗逆转录病毒治疗（cART）。在一些 在实验中，我们还将iPSC衍生的星形胶质细胞并入含有小胶质细胞的脑类器官中， 测试星形胶质细胞在神经保护和神经变性中的作用。我们将进行单细胞基因 暴露于滥用药物、HIV产品和cART的iPSC衍生类器官的表达谱分析， 将采用先进的系统生物学策略来产生可测试的机制假设， neuroHIV中神经变性和组织损伤的发病机制（目的1），并确定候选调节剂 的LTR启动子，可能揭示了对潜伏期和HIV前病毒的再激活的调节（目的 2）。在初步研究中，这种计算实验方法使我们能够确定候选驱动程序， 与神经HIV和神经退行性疾病（包括阿尔茨海默氏症）相关的基因表达变化 疾病、毒品和酒精滥用。这些发现支持了一个总体假设，即解剖的 中枢神经系统基因调控网络的建立将为新的神经元的鉴定铺平道路。 机制假说和药物靶点，以改善PLWHA的神经心理功能， 药物滥用并发症