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: 10672966
• 负责人: Vez REPUNTE-CANONIGO
• 金额: $ 88.83万
• 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/10672966
• 关键词: Alcohol abuse; Alzheimer' s Disease; Anti-Retroviral Agents; Astrocytes; Automobile Driving; CRISPR/Cas technology; Cells; Central Nervous System; 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; HIV/AIDS; Human; Inflammation; Injury; Integration Host Factors; Literature; Macrophage; Mediator; Methamphetamine; Microglia; Modeling; Monitor; Morphine; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Differentiation; 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; Viral; Viral Proteins; Viral reservoir; Virus; advanced system; antiretroviral therapy; candidate identification; 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; 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 感染 HIV 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。在本提案中，我们将使用人类诱导多能干细胞（iPSC）来生成。 皮质和多巴胺能神经元以及小胶质细胞，将被培养为类器官来模拟 HIV 药物 我们展示了与深入的基因组方法和电生理学相结合的滥用相互作用。 电生理学结果证明我们的类器官中神经元的分化水平 与体内和离体制剂相当，是科学界中最好的制剂之一 确定 HIV 表达的候选宿主调节因子和 HIV 诱导的组织损伤的调节因子。 和疾病进展，我们将含有小胶质细胞的大脑类器官暴露于甲基苯丙胺 （冰毒）（一种兴奋剂）和吗啡（一种阿片类药物）属于两类滥用药物， 在艾滋病毒/艾滋病患者 (PLWHA) 中更为普遍 脑类器官会接触有毒物质。 HIV 产品，例如 Tat、gp120、滥用药物和联合抗逆转录病毒疗法 (cART)。 实验中，我们还将把 iPSC 衍生的星形胶质细胞整合到含有小胶质细胞的大脑类器官中，以 我们将进行单细胞基因测试星形胶质细胞在神经保护和神经变性中的作用。 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，我们 将采用先进的系统生物学策略来生成可测试的机制假设 神经HIV中神经变性和组织损伤的发病机制（目标1）并确定候选调节因子 LTR 启动子的研究可能有助于揭示 HIV 原病毒潜伏期和重新激活的调节（Aim 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因素。 与神经艾滋病毒和神经退行性疾病（包括阿尔茨海默病）相关的基因表达变化 这些发现支持了解剖的总体假设。 中枢神经系统的基因调控网络将为识别新型药物铺平道路 改善 PLWHA 和 PLWHA 神经心理功能的机制假设和药物靶点 药物滥用合并症。

项目成果

Maturation Delay of Human GABAergic Neurogenesis in Fragile X Syndrome Pluripotent Stem Cells.

• DOI: 10.1093/stcltm/szac022
• 发表时间: 2022-06-22
• 期刊: STEM CELLS TRANSLATIONAL MEDICINE
• 影响因子: 6
• 作者: Zhang, Ai;Sokolova, Irina;Domissy, Alain;Davis, Joshua;Rao, Lee;Utami, Kagistia Hana;Wang, Yanling;Hagerman, Randi J.;Pouladi, Mahmoud A.;Sanna, Pietro;Boland, Michael J.;Loring, Jeanne F.
• 通讯作者: Loring, Jeanne F.