HIV Tat and morphine-mediated pyroptosis activates astrocytes: Role of NLRP6 inflammasome in HAND

HIV Tat 和吗啡介导的细胞焦亡激活星形胶质细胞：NLRP6 炎性体在 HAND 中的作用

• 批准号: 10217093
• 负责人: Palsamy Periyasamy
• 金额: $ 38.25万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10217093
• 关键词: Astrocytes; Biological Assay; Biological Process; Brain; CASP1 gene; Cell physiology; Cells; Centers for Disease Control and Prevention (U.S.); Confocal Microscopy; DNA; DNA Methylation; DNA sequencing; Data; Development; Disease Progression; Down-Regulation; Drug abuse; Enzyme-Linked Immunosorbent Assay; Enzymes; Epidemic; Epigenetic Process; Exposure to; Family; Future; Gene Expression; Gene Silencing; Genetic Transcription; Genomic DNA; Goals; HIV; HIV Infections; HIV-1; HIV-associated neurocognitive disorder; Health; Homeostasis; Human; IL18 gene; Immunoprecipitation; In Vitro; Individual; Inflammasome; Interleukin-1 beta; Knockout Mice; Knowledge; Lead; Luciferases; Macaca mulatta; Mediating; Mediator of activation protein; MicroRNAs; Microglia; Molecular; Morphine; Morphine Abuse; Mus; Neuraxis; Opioid; Pain management; Pharmaceutical Preparations; Pharmacology; Proteins; Public Health; Reporting; Research; Role; SIV; Signal Transduction; Signaling Protein; Societies; Testing; Tissues; Trans-Activators; Transgenic Organisms; Untranslated RNA; Untranslated Regions; Up-Regulation; Validation; Viral Proteins; Viremia; Western Blotting; antiretroviral therapy; base; bisulfite; bisulfite sequencing; brain tissue; combinatorial; comorbidity; cytokine; cytotoxic; demethylation; drug of abuse; exposed human population; frontal lobe; glial activation; in vivo; lymph nodes; marenostrin; neuroAIDS; neuroinflammation; novel therapeutic intervention; novel therapeutics; opioid use; promoter; receptor; response; therapy adherence; whole genome

Project Summary Despite the ability of combination antiretroviral therapy to dramatically suppress viremia, the brain continues to be a reservoir of HIV low-level replication. Adding further complexity to this is the co-morbidity of drug abuse with HIV-associated neurocognitive disorders and neuroHIV. Among several abused drugs, the use of opiates is highly prevalent in HIV-infected individuals, both as an abused drug as well as for pain management. It has been shown that both HIV/HIV proteins and abused drugs such as morphine contribute to neuroinflammation, which, in turn, involves activation of the inflammasomes. Mounting evidence has also shown that noncoding RNAs function as epigenetic and post-transcriptional regulators controlling vital cellular functions, including activation, thus altering the dynamics of various biological processes. While both HIV proteins and opioids have been shown to change the gene expression profiles of CNS cells, there is a gap of knowledge on the detailed molecular mechanism(s) underlying the co-operative effects of HIV Transactivator of transcription (Tat) protein and morphine on astrocyte activation. The central hypothesis of this proposal is that HIV Tat and morphine, via distinct regulatory mechanism(s) augment astrocyte activation leading, in turn, to exacerbated neuroinflammation. Our preliminary data on whole-genome bisulfite sequencing in the frontal cortices of SIV- infected rhesus macaques show increased DNA hypomethylation of the NLRP6 (NOD-like receptor family, pyrin domain-containing protein 6) promoter with a concomitant upregulation in NLRP6 expression. Additionally, exposure of human and mouse primary astrocytes to HIV Tat and morphine also resulted in decreased expression of microRNA-152 that was accompanied by increased expression of NLRP6. Interestingly, we also found that exposure of these primary astrocytes to either HIV Tat or morphine resulted in increased cellular activation with increased expression of proinflammatory cytokines (IL1β and IL18) via pyroptosis. Based on the central hypothesis and the reliable preliminary data, this proposal led to the following specific aims: Specific Aim 1: Determine the molecular mechanism(s) involved in microRNA-152 mediated NLRP6 inflammasome activation in HIV Tat and morphine-exposed astrocytes in vitro; Specific Aim 2: Determine the epigenetic mechanism(s) involved in promoter DNA hypomethylation of the NLRP6 in HIV Tat and morphine-exposed astrocytes in vitro; Specific Aim 3: Validate the combinatorial effects of HIV Tat and morphine on NLRP6 inflammasome mediated astrocyte activation, in vivo. Understanding the mechanisms responsible for astrocyte activation induced by HIV and morphine will set the stage for the future development of novel therapeutics aimed at dampening HIV and opiate-mediated neuroinflammatory responses.

项目概要 尽管联合抗逆转录病毒疗法能够显着抑制病毒血症，但大脑仍继续 是HIV低水平复制的储存库。使这一问题变得更加复杂的是药物滥用与 HIV 相关的神经认知障碍和神经 HIV。在多种滥用药物中，阿片类药物的使用最为严重 在艾滋病毒感染者中非常普遍，既可以作为滥用药物，也可以用于止痛。它一直 研究表明，HIV/HIV 蛋白和吗啡等滥用药物都会导致神经炎症， 反过来，涉及炎症小体的激活。越来越多的证据还表明，非编码 RNA 作为表观遗传和转录后调节因子控制重要的细胞功能，包括激活、 从而改变各种生物过程的动态。虽然 HIV 蛋白和阿片类药物都已被证明 为了改变中枢神经系统细胞的基因表达谱，在详细的分子分子方面存在知识空白 HIV 转录反式激活蛋白 (Tat) 协同作用的潜在机制 吗啡对星形胶质细胞激活的影响。该提案的中心假设是 HIV Tat 和吗啡，通过 不同的调节机制增强星形胶质细胞的激活，进而导致加剧 神经炎症。我们关于 SIV 额叶皮质全基因组亚硫酸氢盐测序的初步数据- 受感染的恒河猴表现出 NLRP6（NOD 样受体家族、pyrin 包含结构域的蛋白 6) 启动子伴随着 NLRP6 表达的上调。此外， 人类和小鼠原代星形胶质细胞暴露于 HIV Tat 和吗啡也导致 microRNA-152 的表达伴随着 NLRP6 表达的增加。有趣的是，我们还 发现这些初级星形胶质细胞暴露于 HIV Tat 或吗啡会导致细胞 通过细胞焦亡增加促炎细胞因子（IL1β 和 IL18）表达的激活。基于 中心假设和可靠的初步数据，该提案导致了以下具体目标： 具体目标 1: 确定 microRNA-152 介导的 NLRP6 炎性体激活的分子机制 HIV Tat 和吗啡暴露的星形胶质细胞体外；具体目标 2：确定表观遗传机制 参与体外 HIV Tat 和吗啡暴露的星形胶质细胞中 NLRP6 启动子 DNA 的低甲基化； 具体目标 3：验证 HIV Tat 和吗啡对 NLRP6 炎症小体介导的组合作用 体内星形胶质细胞激活。了解 HIV 诱导星形胶质细胞激活的机制 吗啡将为未来开发旨在抑制艾滋病毒和艾滋病毒的新型疗法奠定基础 阿片介导的神经炎症反应。