Molecular Pathways of Innate Immunity and Substance Abuse in NeuroHIV

NeuroHIV 的先天免疫和药物滥用的分子途径

• 批准号: 10814534
• 负责人: Joao Filipe Inacio Mamede
• 金额: $ 42.34万
• 依托单位: RUSH UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10814534
• 关键词: Animal Model; Anti-Retroviral Agents; Antigen-Antibody Complex; Astrocytes; Autopsy; Binding Proteins; Biological; Biological Assay; Biological Markers; Brain; CRISPR/Cas technology; Capsid; Cell Culture Techniques; Cerebrum; Cessation of life; Chronic; Clinical; Complex; Consequences of HIV; Cyclic GMP; Cytokine Signaling; DNA Damage; Data; Dendritic Cells; Detection; Development; Drug abuse; Event; Exhibits; Fluorescence Microscopy; Functional disorder; Future; Gene Activation; Gene Expression; Genomics; HIV; HIV Infections; HIV Seronegativity; HIV therapy; HIV-1; HIV-associated neurocognitive disorder; HIV/AIDS; Heart; High Prevalence; Homeostasis; Human; IRF3 gene; Image; Immunofluorescence Immunologic; Impaired cognition; Individual; Inflammation; Inflammatory; Interferons; Knowledge; Machine Learning; Measures; Mediating; Methamphetamine; Microglia; Microscopy; Modeling; Modification; Molecular; Morbidity - disease rate; Mus; Myeloid Cell Activation; Natural Immunity; Neurocognitive Deficit; Neuroglia; Neurons; Organoids; Outcome Assessment; Pathogenicity; Pathway interactions; Pattern; Pattern recognition receptor; Periodicity; Persons; Pharmaceutical Preparations; Phase; Phenotype; Physiological; Play; Process; Production; Research; Reverse Transcription; Role; Sampling; Signal Transduction; Stimulator of Interferon Genes; Substance abuse problem; Techniques; Therapeutic; Time; Viral; abuse victim; antagonist; antiretroviral therapy; brain cell; comorbidity; cytokine; experience; fluorescence imaging; improved; induced pluripotent stem cell; innate immune sensing; long term abstinence; methamphetamine abuse; methamphetamine effect; methamphetamine use; methamphetamine user; microscopic imaging; neuroAIDS; neurocognitive disorder; neuroinflammation; neurotropic; next generation sequencing; particle; pathogen; polyglutamine; receptor; repaired; single-cell RNA sequencing; targeted treatment; ultra high resolution

PROJECT SUMMARY A significant percentage of people living with HIV/AIDS (PLWH) exhibit HIV associated neurocognitive disorders (HAND). With the development and improvement of combination antiretroviral therapies (cART) there was a substantial decrease of the death and morbidity rates of PLWH. HAND and neuroinflammation are exacerbated by Methamphetamine (Meth) abuse. PLWH abusing Meth present a high prevalence of chronic neuroinflammation and the cellular mechanism(s) that drive such inflammation have not been entirely defined. Certain models of neuroinflammation define inter-cellular cytokine signaling as a major player in neuroinflammation. These models propose that activation of microglia results in the production of inflammatory cytokines and metabolites that are recognized by astrocytes, leading to a modification from non-reactive astrocytes to a range of pro-inflammatory or repair phenotypes. In this proposal we aim to identify the molecular mechanisms that trigger the activation of intracellular innate immunity in microglia and astrocytes in the context of NeuroHIV and Meth, and characterize the cellular crosstalk between HIV infected and non-infected brain cells under physiological concentrations of cART. Our preliminary data from gene expression and microscopy of innate sensing immune complexes points to an activation of the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway in microglia with Meth and HIV infection. Thus, we hypothesize that HIV-triggered activation of the cGAS/STING pathway leads to episodes of chronic neuroinflamation. We also hypothesize that Meth-mediated DNA damage is additive to the activation of the inflammation pathway. In the R61 phase, we aim to determine the impact of HIV post-entry events and Meth-mediated DNA damage on innate immune sensing of glial cells. During the R33 phase, we will characterize the biological consequences of HIV/Meth-mediated innate sensing to brain organoids under brain physiologic cART concentrations. We aim to assess the potential of cGAS/STING antagonists to reduce neuroinflammation in the context of HIV/Meth. We will carry out an experimental plan using functional assays, cyclic multiplex immunofluorescence and super-resolution fluorescence imaging, machine learning analysis, next-generation sequencing, and multiplexed cytokine profiling in iPSC-derived microglia, astrocytes, cerebral brain organoids and a chimeric human/mouse animal model. These studies will define the activation of HIV/Meth-mediated innate immunity in cells from the brain that are known to be infected by HIV and the impact of this activation in the disruption of normal brain homeostasis by inflammatory processes. At the heart of this proposal is the clarification and identification of the mechanism(s) that drive and/or contribute to persistent neuroinflammation by HIV and Meth under cART that culminate in HAND. Knowledge gained from this research plan can inform therapeutic potential to ameliorate and/or reduce HAND in PLWH.

项目摘要 很大一部分艾滋病毒/艾滋病感染者（PLWH）表现出与艾滋病毒相关的神经认知功能障碍。 疾病（手）。随着联合抗逆转录病毒疗法（cART）的发展和完善， 艾滋病毒/艾滋病感染者的死亡率和发病率大幅下降。手和神经炎症是 滥用甲基苯丙胺（Methamphetamine）。滥用甲基苯丙胺的艾滋病毒携带者慢性 神经炎症和驱动这种炎症的细胞机制尚未完全确定。 某些神经炎症模型将细胞间细胞因子信号传导定义为神经炎症的主要参与者。 神经炎症这些模型表明，小胶质细胞的激活导致炎症反应的产生。 细胞因子和代谢物，由星形胶质细胞识别，导致从非反应性的修饰， 星形胶质细胞的一系列促炎或修复表型。在这项提案中，我们的目标是确定分子 在小胶质细胞和星形胶质细胞中触发细胞内先天免疫激活的机制 的NeuroHIV和甲基，并表征HIV感染和未感染的脑细胞之间的细胞串扰 在生理浓度的cART下。 我们从先天感应免疫复合物的基因表达和显微镜检查中获得的初步数据表明， 环GMP-AMP合酶（cGAS）/干扰素基因刺激物（STING）途径的激活 小胶质细胞与甲基苯丙胺和艾滋病毒感染。因此，我们假设HIV触发的cGAS/STING激活 这一通路导致慢性神经炎症发作。我们还假设甲基化酶介导的DNA损伤 对炎症通路的激活是附加的。在R61阶段，我们的目标是确定 HIV进入后事件和Meth-mediated DNA损伤对神经胶质细胞先天免疫感应的影响在R33 阶段，我们将描述HIV/Meth-mediated先天感知对脑类器官的生物学后果 在脑生理cART浓度下。我们的目标是评估cGAS/STING拮抗剂的潜力， 减少HIV/Meth背景下的神经炎症。我们将使用函数进行实验计划 检测、循环多重免疫荧光和超分辨率荧光成像、机器学习 iPSC衍生的小胶质细胞，星形胶质细胞， 脑类器官和嵌合人/小鼠动物模型。 这些研究将确定HIV/Met介导的先天免疫在大脑细胞中的激活， 已知被HIV感染的人，以及这种激活对正常脑内稳态的破坏的影响， 炎症过程。这项建议的核心是澄清和确定 在cART下驱动和/或促成HIV和甲氧西林持续性神经炎症的机制 这是在手。从这项研究计划中获得的知识可以为治疗潜力提供信息， 改善和/或减少PLWH中的HAND。