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

摘要 METH是一种强效成瘾性精神兴奋剂，是美国最常滥用的药物之一。 甲基苯丙胺滥用在艾滋病毒感染者中非常普遍，这对艾滋病毒提出了独特的挑战 预防和治疗。鉴于METH使用和HIV对CNS神经元损伤的重叠影响， 迫切需要了解METH和HIV在HIV发病机制中的相互作用- 相关的神经认知障碍（HAND）。然而，HAND的研究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元细胞。最近成功地在 从人类诱导多能干细胞系（iPSC）中产生小胶质细胞和神经元现在提供了一个很好的 研究HIV感染的小胶质细胞和神经元之间的直接相互作用的机会。拟议的研究 将使用iPSC衍生的小胶质细胞（iPSC-MG）和神经元（iPSC-N）来检验我们的总体假设，即METH 和/或HIV抑制细胞内HIV限制因子并诱导炎性小体和神经毒性 iPSC-MG中的miRNA，其促进iPSC-MG中的HIV感染/复制并促进iPSC-N的死亡。 我们提出两个具体目标来解决这个假设： 目的1确定METH是否增强iPSC-MG中的HIV感染/复制。机械地，我们将 检查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METH和/或HIV感染是否诱导炎性小体的表达， 神经毒性miRNA在iPSC-MG中的作用并促进IPSC-N的死亡。 该项目非常符合NIDA最近对研究药物滥用和/或艾滋病毒对人类的影响的兴趣。 iPSC衍生的CNS细胞。使用iPSC-MG和iPSC-N的拟议研究所产生的信息 将提供重要的见解如何相互作用的两个主要病理因素（艾滋病毒和甲基）在 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 HIV感染/在大脑中的持续存在和HAND的发育机制。

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.