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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要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能力疾病（手）。随着联合抗逆转录病毒疗法（cART）的发展和完善 PLWH 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（Meth）会加剧这种情况。艾滋病毒感染者滥用冰毒导致慢性病患病率很高神经炎症和驱动此类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主要参与者神经炎症。这些模型提出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，导致非反应性的修饰星形胶质细胞具有一系列促炎或修复表型。在本提案中，我们的目标是确定分子在背景下触发小胶质细胞和星形胶质细胞细胞内先天免疫激活的机制 NeuroHIV 和 Meth 的研究，并表征 HIV 感染和未感染脑细胞之间的细胞串扰在cART的生理浓度下。我们从先天传感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)/干扰素基因刺激剂 (STING) 通路的激活感染冰毒和艾滋病毒的小胶质细胞。因此，我们假设 HIV 触发的 cGAS/STING 激活途径导致慢性神经炎症发作。我们还假设冰毒介导的 DNA 损伤有助于炎症途径的激活。在 R61 阶段，我们的目标是确定 HIV 进入后事件和冰毒介导的 DNA 损伤对神经胶质细胞的先天免疫感应。 R33期间阶段，我们将描述 HIV/Me 介导的先天感知对大脑类器官的生物学后果在大脑生理cART浓度下。我们的目的是评估 cGAS/STING 拮抗剂的潜力减少艾滋病毒/冰毒背景下的神经炎症。我们将使用函数式进行实验计划分析、循环多重免疫荧光和超分辨率荧光成像、机器学习对 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。这些研究将确定大脑细胞中 HIV/冰毒介导的先天免疫的激活。已知被 HIV 感染，以及这种激活对正常大脑稳态破坏的影响炎症过程。该提案的核心是澄清和确定 cART 下 HIV 和冰毒驱动和/或促成持续性神经炎症的机制最终在 HAND 中达到顶峰。从该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。