Methamphetamine and Antiretroviral Therapy Impact Macrophage Functions and Macroautophagy: Implications for HIV Neuropathogenesis

甲基苯丙胺和抗逆转录病毒治疗影响巨噬细胞功能和巨自噬：对 HIV 神经发病机制的影响

• 批准号: 10456092
• 负责人: John M Barbaro
• 金额: $ 5.18万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10456092
• 关键词: Address; Affect; Amyloid beta-Protein; Astrocytes; Autophagocytosis; Autophagosome; Brain; Catabolic Process; Cell physiology; Cells; Chronic; Development; Drug usage; Enzyme-Linked Immunosorbent Assay; Extracellular Protein; Flow Cytometry; Fluorescence Microscopy; Fluorometry; Gene Expression; Glutamates; Goals; Growth Factor; HIV; HIV-associated neurocognitive disorder; Health; Homeostasis; Human; Impairment; Incidence; Infection; Inflammasome; Inflammatory; Interleukin-1; Interleukin-18; Intervention; Knowledge; Life; Lysosomes; Mediating; Methamphetamine; Microglia; Microscopy; Mitochondria; Myeloid Cells; Nervous System Trauma; Neurocognitive Deficit; Neurologic; Neurons; Neuropathogenesis; Organelles; Oxidative Stress; Peripheral; Persons; Phagocytes; Phagocytosis; Pharmaceutical Preparations; Phenotype; Process; Production; Proteins; Quality Control; Quality of life; Reactive Oxygen Species; Receptor Signaling; Regimen; Role; Severities; Signal Transduction; Substance Use Disorder; System; Time; Toll-like receptors; Viral reservoir; Western Blotting; antiretroviral therapy; cell injury; cell type; comorbidity; cytokine; excitotoxicity; experimental study; extracellular; high risk; macrophage; metabolomics; methamphetamine effect; methamphetamine use; methamphetamine user; monocyte; mortality; neurotoxic; neurotransmitter metabolism; neurotransmitter release; pathogen; protein aggregation; relating to nervous system; response; therapy development; transmission process

Project Abstract Myeloid cells, microglia and monocyte derived macrophages (MDM), are long-lived cellular reservoirs for HIV in the CNS not eliminated by effective antiretroviral therapy (ART). Despite ART, infected cells become activated and damaged, leading to the development of HIV-associated neurocognitive disorders, or HAND. Approximately 15-40% of people living with HIV (PLWH) have some form of HAND despite ART. Methamphetamine (meth) use can facilitate HIV transmission and may increase CNS damage in meth users, including PLWH. We will characterize how meth impacts microglia/MDM mediated inflammatory processes that contribute to CNS damage in PLWH taking ART. We will also determine how changes in these cellular functions in response to meth and ART may be regulated by macroautophagy. Macroautophagy is a homeostatic, catabolic process in which damaged and toxic proteins, organelles, and pathogens are targeted for cytosolic degradation in the lysosome. Some studies have shown that macroautophagy may, in part, regulate functions of microglia/MDM that contribute to CNS damage and HAND. We hypothesize that meth and ART impact autophagy in microglia/MDM, leading to impaired clearance of extracellular debris and increased neurotoxic ROS, cytokine, and glutamate secretion to contribute to neurological damage in people taking meth and ART. To study this hypothesis, we developed two aims: Aim 1: To determine the impact of meth and ART on cellular functions of uninfected and HIV-infected human microglia and monocyte derived macrophages (MDM) that contribute to HAND. We will determine how meth and ART impact phagocytosis, ROS production, cytokine secretion, and glutamate release using fluorometry, microscopy, flow cytometry, and ELISA. Aim 2: To characterize how meth and ART impact macroautophagy in uninfected and HIV- infected human microglia and monocyte derived macrophages (MDM), and determine how macroautophagy regulates meth and ART mediated changes in functions that contribute to HAND. We examine how meth and ART change general and selective autophagic processes using high-content microscopy and Western blotting. We will also genetically inhibit macroautophagy or use drugs that inhibit or activate autophagy to determine further how autophagy may regulate meth and ART induced changes in phagocytosis, ROS, cytokine secretion, and glutamate release. With this knowledge, we can develop therapies that impact macroautophagy to mitigate microglial/MDM mediated CNS damage, reducing HAND in meth users living with HIV.

项目摘要 骨髓细胞、小胶质细胞和单核细胞衍生的巨噬细胞 (MDM) 是 HIV 的长寿命细胞库 有效的抗逆转录病毒疗法（ART）无法消除中枢神经系统中的病毒。尽管接受了 ART，受感染的细胞仍会变得 激活和受损，导致 HIV 相关神经认知障碍（HAND）的发展。 尽管接受了 ART，但大约 15-40% 的 HIV 感染者 (PLWH) 仍患有某种形式的 HAND。 使用甲基苯丙胺（甲基苯丙胺）可促进艾滋病毒传播，并可能增加甲基苯丙胺使用者的中枢神经系统损伤， 包括艾滋病病毒感染者。我们将描述冰毒如何影响小胶质细胞/MDM 介导的炎症过程， 导致接受抗逆转录病毒疗法的感染者的中枢神经系统损伤。我们还将确定这些细胞的变化如何 对甲基苯丙胺和抗逆转录病毒疗法的反应功能可能受到巨自噬的调节。巨自噬是一种 稳态、分解代谢过程，其中受损和有毒的蛋白质、细胞器和病原体成为目标 用于溶酶体中的胞质降解。一些研究表明，巨自噬可能在一定程度上， 调节小胶质细胞/MDM 的功能，从而导致 CNS 损伤和 HAND。我们假设冰毒和 ART 影响小胶质细胞/MDM 中的自噬，导致细胞外碎片清除受损并增加 神经毒性 ROS、细胞因子和谷氨酸分泌会导致服用冰毒的人出现神经损伤 和艺术。为了研究这一假设，我们制定了两个目标： 目标 1：确定冰毒和毒品的影响 ART 对未感染和 HIV 感染的人类小胶质细胞和单核细胞来源的细胞功能的影响 巨噬细胞 (MDM) 有助于 HAND。我们将确定冰毒和 ART 如何影响吞噬作用， 使用荧光测定法、显微镜、流式细胞术和 酶联免疫吸附试验。目标 2：描述冰毒和抗逆转录病毒疗法如何影响未感染者和 HIV 感染者的巨自噬 感染人类小胶质细胞和单核细胞衍生巨噬细胞 (MDM)，并确定如何 巨自噬调节甲基苯丙胺和 ART 介导的功能变化，从而促进 HAND。我们 研究冰毒和 ART 如何使用高内涵改变一般和选择性自噬过程 显微镜和蛋白质印迹。我们还将从基因上抑制巨自噬或使用抑制或 激活自噬，进一步确定自噬如何调节冰毒和 ART 诱导的变化 吞噬作用、ROS、细胞因子分泌和谷氨酸释放。有了这些知识，我们就可以开发治疗方法 影响巨自噬以减轻小胶质细胞/MDM介导的中枢神经系统损伤，减少甲基苯丙胺中的HAND 感染艾滋病毒的用户。