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

摘要 甲基苯丙胺（METH）是一种强效成瘾性精神兴奋剂，是世界上最常见的滥用药物之一。 美国的在艾滋病毒感染者中，甲基苯丙胺滥用非常普遍，这表现出独特的 艾滋病毒预防和治疗的挑战。鉴于甲基苯丙胺的使用和艾滋病毒对神经元的重叠影响， 在中枢神经系统（CNS）的损害，它成为迫切需要了解的作用之间的相互作用， METH和HIV在HIV相关神经认知障碍（HAND）发病机制中的作用然而，研究 HAND一直受到从HIV患者尸检或活检中收集活脑细胞的困难的阻碍。 从人诱导多能干细胞产生小胶质细胞和脑类器官的最新成功 iPSCs的研究为研究METH和/或HIV对CNS的影响提供了很好的机会。整体 本项目的目的是验证和建立临床相关的体外脑模型， 药物滥用背景下的神经艾滋病研究。该项目将由两名私人侦探进行， 在拟议的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（METH和阿片类药物）对外周/中枢神经系统先天免疫和艾滋病毒感染的影响 巨噬细胞/小胶质细胞。胡博士的研究重点是神经发生，iPSC的产生， 炎性小体、CRISPR/Cas HIV根除和HAND。他们最近的合作成功地 产生并表征了人iPSC衍生的含有小胶质细胞的脑类器官（MC 0）， 表达主要的CNS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 重要的是，这些MCO可能会感染HIV。基于这些重要的发现，我们提出了三个 具体目标：目标1。研究人iPSC衍生的MCO的动态HIV感染和METH的影响 艾滋病毒感染的MCO;目的2.为了确定HIV感染的细胞类型和主要的靶/储库细胞， MCO中的艾滋病毒;目标3。研究HIV感染对MCO发育和神经回路的影响。 本项目的成功完成将为研究艾滋病毒感染的作用提供一个体外脑模型 和滥用药物在神经艾滋病发病机制中的作用。

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.