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: 10645059
• 负责人: John M Barbaro
• 金额: $ 5.27万
• 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/10645059
• 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; IL18 gene; Impairment; Incidence; Infection; Inflammasome; Inflammatory; Intervention; Knowledge; Life; Lysosomes; Macrophage; Mediating; Methamphetamine; Microglia; Microscopy; Mitochondria; Myeloid Cells; Nervous System Trauma; Neurocognitive Deficit; Neurologic; Neurons; Neuropathogenesis; Organelles; Oxidative Stress; Peripheral; Persons; Phagocytes; Phagocytosis; Phagocytosis Inhibition; 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; metabolomics; methamphetamine effect; methamphetamine use; methamphetamine user; monocyte; mortality; neural; neurotoxic; neurotransmitter metabolism; neurotransmitter release; pathogen; protein aggregation; 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 髓样细胞、小胶质细胞和单核细胞衍生的巨噬细胞（MDM）是HIV的长寿细胞库 在CNS中，有效的抗逆转录病毒治疗（ART）不能消除。尽管ART，感染的细胞成为 激活和受损，导致艾滋病毒相关的神经认知障碍，或手的发展。 大约15-40%的艾滋病毒感染者（PLWH）尽管有抗逆转录病毒治疗，但仍有某种形式的手。 甲基苯丙胺（冰毒）的使用可以促进艾滋病毒的传播，并可能增加冰毒使用者的中枢神经系统损伤， 包括PLWH。我们将描述冰毒如何影响小胶质细胞/MDM介导的炎症过程， 有助于中枢神经系统的损害PLWH服用艺术。我们还将确定如何改变这些细胞 对甲氨蝶呤和抗逆转录病毒疗法的应答功能可能受到巨自噬的调节。自噬是一种 一种以受损和有毒蛋白质、细胞器和病原体为目标的自我平衡、分解代谢过程 用于溶酶体中的胞质降解。一些研究表明，巨自噬可能，部分， 调节导致CNS损伤和HAND的小胶质细胞/MDM的功能。我们假设冰毒和 ART影响小胶质细胞/MDM中的自噬，导致细胞外碎片的清除受损， 神经毒性ROS、细胞因子和谷氨酸分泌导致服用冰毒的人神经损伤 为了研究这一假设，我们制定了两个目标：目标1：确定冰毒的影响， ART对未感染和HIV感染的人小胶质细胞和单核细胞功能的影响 巨噬细胞（MDM），有助于手。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、流式细胞术和流式细胞仪检测ROS产生、细胞因子分泌和谷氨酸释放， ELISA法目的2：描述meth和ART如何影响未感染和HIV-1感染者的巨自噬。 感染的人小胶质细胞和单核细胞衍生的巨噬细胞（MDM），并确定如何 巨细胞自噬调节METH和ART介导的有助于HAND的功能变化。我们 研究meth和ART如何使用高含量的药物改变一般和选择性自噬过程 显微镜和Western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，以进一步确定自噬如何调节甲基苯丙胺和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CNS损伤，减少方法中的HAND 艾滋病毒感染者。