Effect of Methamphetamine and/or HIV on Human iPSCs-derived microglia and Neuron

摘要 冰毒是一种有效的成瘾精神刺激剂，是美国最常见的滥用药物之一。 吸食冰毒在艾滋病毒感染者中非常普遍，这对艾滋病毒构成了独特的挑战 预防和治疗。鉴于冰毒和艾滋病毒对中枢神经系统神经元损伤的重叠影响，它 迫切需要了解冰毒和艾滋病毒之间的相互作用在艾滋病毒发病机制中的作用 相关神经认知障碍(手)。然而，手部的研究一直受到以下困难的阻碍 从HIV患者的尸检或活检中收集原代小胶质细胞和神经细胞。最近在以下方面取得了成功 从人类诱导的多能干细胞系(IPSCs)中培养小胶质细胞和神经元现在提供了一种很好的 有机会研究艾滋病毒感染的小胶质细胞与神经元之间的直接相互作用。建议进行的研究 将使用IPSC来源的小胶质细胞(IPSC-MG)和神经元(IPSC-N)来检验我们的总体假设 和/或HIV抑制细胞内HIV限制因子，诱导炎症小体和神经毒性 在iPSC-MG中，miRNAs促进了HIV在iPSC-MG中的感染/复制，并促进了iPSC-N的死亡。 我们提出了两个具体目标来解决这一假设： 目的1确定冰毒是否能增强IPSC-MG中的HIV感染/复制。从机制上讲，我们会 检测METH和/或HIV是否抑制IPSC-MG中的细胞内HIV限制因子； 目的2确定冰毒和/或HIV感染是否诱导炎性小体表达和 神经毒性miRNAs在IPSC-MG中的表达，并促进IPSC-N的死亡。 这个项目非常符合NIDA最近在研究药物滥用和/或艾滋病毒对人类的影响方面的兴趣 IPSC来源的中枢神经系统细胞。使用IPSC-MG和IPSC-N进行的拟议研究得出的信息 将提供关于两种主要病理因素(艾滋病毒和冰毒)如何在人类免疫缺陷病毒中相互作用的重要见解 中枢神经系统损害小胶质细胞的细胞内抗HIV免疫，诱导神经元死亡，这是关键 艾滋病毒感染/持续存在于大脑和手发育的机制。

Abstract 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 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 primary microglial and neuronal cells from autopsy or biopsy of HIV patients. Recent success in generating microglia and neurons from human induced pluripotent stem cell lines (iPSCs) now offers a great opportunity to study the direct interactions between HIV-infected microglia and neurons. The proposed studies will use iPSC-derived microglia (iPSC-MG) and neuron (iPSC-N) to examine our overall hypothesis that METH and/or HIV inhibit the intracellular HIV restriction factors and induce the inflammasomes and neurotoxic miRNAs in iPSC-MG, which facilitate HIV infection/replication in iPSC-MG and promote the death of iPSC-N. We propose two specific aims to address this hypothesis: Aim 1 To determine whether METH enhances HIV infection/replication in iPSC-MG. Mechanistically, we will examine whether METH and/or HIV inhibit the intracellular HIV restriction factors in iPSC-MG; Aim 2 To determine whether METH and/or HIV infection induce expression of the inflammasomes and neurotoxic miRNAs in iPSC-MG and promote the death of IPSC-N. This project fits well to the NIDA’s recent interests in studying the impact of drug abuse and/or HIV on human iPSC-derived CNS cells. The information resulting from the proposed studies using the iPSC-MG and iPSC-N will provide important insights on how the interplays of two major pathologic factors (HIV and METH) in the CNS compromise the intracellular anti-HIV immunity of microglia and induce neuronal death, which are the key mechanisms for HIV infection/persistence in the brain and development of HAND.