HIV, Methamphetamine and Human iPSC-derived Microglia-containing Cerebral Organoids

HIV、甲基苯丙胺和人 iPSC 衍生的含有小胶质细胞的大脑类器官

• 批准号: 10205018
• 负责人: WENZHE HO
• 金额: $ 61.76万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10205018
• 关键词: AIDS prevention; Acute; Astrocytes; Autopsy; Biological Assay; Biological Models; Biopsy; Brain; Cell Line; Cells; Cellular Structures; Cerebrum; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; DLG4 gene; DNA; Development; Drug abuse; Enzyme-Linked Immunosorbent Assay; Feasibility Studies; Flow Cytometry; GAG Gene; Generations; Glial Fibrillary Acidic Protein; Growth; HIV; HIV Infections; HIV-associated neurocognitive disorder; Human; ITGAM gene; Immunohistochemistry; In Vitro; Individual; Inflammasome; MCM2 gene; Methamphetamine; Methods; Microglia; Modeling; Molecular; National Institute of Drug Abuse; Natural Immunity; Nature; Neuraxis; Neurons; Opioid; Organ; Organoids; Pathogenesis; Patients; Peripheral; Physiological; Protocols documentation; RNA; Reproducibility; Research; Role; Synaptophysin; Time; United States; Virus; Western Blotting; Work; acute infection; base; brain cell; cell type; chronic infection; clinically relevant; confocal imaging; drug of abuse; experience; human model; induced pluripotent stem cell; interest; macrophage; methamphetamine abuse; methamphetamine effect; methamphetamine use; methamphetamine user; nervous system development; nestin protein; neuroAIDS; neurogenesis; neuron apoptosis; neuronal circuitry; neuronal survival; neurotropic; psychostimulant; relating to nervous system; stem; stem cells; success; synaptogenesis; synaptotagmin

Abstract Methamphetamine (METH), a potent addictive psychostimulant, is one of the most commonly abused drugs in the United States. METH abuse is highly prevalent in HIV-infected individuals, which presents unique challenges for HIV prevention and treatment. Given the overlap impact of METH use and HIV on neuronal damage in the central nervous system (CNS), it becomes urgent to understand the role of interplays between METH and HIV in the pathogenesis of HIV-associated neurocognitive disorders (HAND). However, studies of HAND have been hampered by difficulties in collecting live brain cells from autopsy or biopsy of HIV patients. Recent success in generating microglia and cerebral organoids from human induced pluripotent stem cells (iPSCs) now offers a great opportunity to investigate the impact of METH and/or HIV on the CNS. The overall objective of this project is to validate and establish a clinically relevant in vitro brain model for NeuroAIDS research in the context of drug abuse. The project will be carried out by two P.I.s who have the complementary expertise and experience in the proposed studies. Dr. Ho has been working on the impact of abused drugs (METH and opioids) on peripheral/CNS innate immunity and HIV infection of macrophages/microglia since 1999. Dr. Hu’s research focuses on neurogenesis, iPSC generation, inflammasomes, CRISPR/Cas HIV eradication and HAND. Their recent collaborative work has successfully generated and characterized the human iPSC-derived microglia-containing cerebral organoids (MCOs) which express the major CNS cell types: neural stem/progenitor cells, neurons, astrocytes, and microglia. More importantly, these MCOs could be infected by HIV. Based on these important findings, we propose three specific aims: Aim 1. To study dynamic HIV infection of human iPSC-derived MCOs and the impact of METH on HIV infection of MCOs; Aim 2. To determine HIV-infected cell types and the primary target/reservoir cells of HIV in MCOs; Aim 3. To study the impact of HIV infection on the development and neuronal circuitry in MCOs. Successful completion of this project should provide an in vitro brain model to study the roles of HIV infection and drugs of abuse in the pathogenesis of NeuroAIDS.

摘要 甲基苯丙胺(甲基苯丙胺)是一种有效的成瘾精神刺激剂，是全球最常见的滥用药物之一 美国。吸食冰毒在艾滋病毒感染者中非常普遍，这表现出独特的 艾滋病毒预防和治疗面临的挑战。鉴于冰毒使用和艾滋病毒对神经元的重叠影响 在中枢神经系统(CNS)受损的情况下，迫切需要了解 冰毒和HIV在HIV相关神经认知障碍(HAND)发病机制中的作用。然而，对 由于难以从艾滋病毒患者的身体解剖或活组织检查中收集活的脑细胞，手部一直受到阻碍。 人诱导多能干细胞分化为小胶质细胞和类脑器官的研究进展 (IPSCs)现在提供了一个很好的机会来研究冰毒和/或艾滋病毒对中枢神经系统的影响。整体而言 本项目的目的是验证和建立一种具有临床意义的体外脑模型 药物滥用背景下的神经艾滋病研究。该项目将由两名私家侦探执行，他们拥有 在拟议研究中补充专门知识和经验。何博士一直致力于研究 滥用药物(冰毒和阿片类药物)对外周/中枢神经系统天然免疫和HIV感染的影响 巨噬细胞/小胶质细胞自1999年以来。胡博士的研究重点是神经发生，IPSC的产生， 炎症体，CRISPR/Cas HIV根除和Hand。他们最近的合作工作取得了成功 产生并鉴定了人IPSC来源的含有小胶质细胞的脑器质(MCOs)，它 表达主要的中枢神经系统细胞类型：神经干/祖细胞、神经元、星形胶质细胞和小胶质细胞。更多 重要的是，这些MCO可能会感染艾滋病毒。基于这些重要的发现，我们提出了三点 目的：1.研究人IPSC来源的MCOs感染HIV的动态变化及冰毒对其的影响 目的2.确定人类免疫缺陷病毒感染的细胞类型和主要的靶/储存细胞。 目的3.研究HIV感染对MCOs发育和神经元回路的影响。 该项目的成功完成将为研究HIV感染的作用提供一个体外脑模型 以及药物滥用在神经艾滋病发病机制中的作用。