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.

项目总结 相当大比例的艾滋病毒/艾滋病患者(PLWH)表现出与艾滋病毒相关的神经认知 精神障碍(手)。随着联合抗逆转录病毒疗法(CART)的发展和完善 是PLWH死亡率和发病率的大幅下降。手部和神经性炎症 甲基苯丙胺(Meth)的滥用加剧了这一现象。PLWH滥用Meth呈现慢性高患病率 神经炎症和驱动这种炎症的细胞机制(S)还没有完全确定。 某些神经炎症模型将细胞间细胞因子信号转导定义为 神经炎。这些模型表明，小胶质细胞的激活导致炎症的产生。 星形胶质细胞识别的细胞因子和代谢物，导致非反应性修饰 星形胶质细胞具有一系列促炎或修复表型。在这项提案中，我们的目标是识别分子 小胶质细胞和星形胶质细胞激活细胞内天然免疫的机制 NeuroHIV和Meth，并表征感染了HIV和未感染的脑细胞之间的细胞串扰 在生理浓度的Cart下。 我们的基因表达和先天感知免疫复合体显微镜的初步数据表明 环状GMP-AMP合成酶(CGAS)/干扰素基因刺激物(STING)通路的激活 Meth和HIV感染的小胶质细胞。因此，我们假设HIV触发了cGAS/STIN的激活 通路导致慢性神经炎症的发作。我们还假设甲基苯丙氨酸介导的DNA损伤 对炎症途径的激活是相加的。在R61阶段，我们的目标是确定 HIV进入后事件和Meth介导的DNA损伤对神经胶质细胞天然免疫感觉的影响。在R33期间 阶段，我们将表征HIV/Meth介导的对脑器官的先天感知的生物学后果 在脑部生理车浓度下。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。我们将执行一项实验计划，使用功能 检测、循环多重免疫荧光和超分辨率荧光成像、机器学习 IPSC来源的小胶质细胞、星形胶质细胞、 脑有机体和人/鼠嵌合动物模型。 这些研究将定义来自大脑的细胞中HIV/Meth介导的天然免疫的激活 已知感染艾滋病毒以及这种激活在破坏正常大脑稳态中的影响 炎症过程。这项提议的核心是澄清和确定 HIV和Meth在CART下驱动和/或促成持续性神经炎的机制(S) 这在手中达到了顶峰。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中的手。