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.

项目摘要 尽管结合抗逆转录病毒疗法能够极大地抑制病毒血症，但大脑仍在继续 成为艾滋病毒低级复制的水库。对此增加了进一步的复杂性 与HIV相关的神经认知障碍和神经hiv。在几种滥用药物中，鸦片使用者的使用是 在艾滋病毒感染的个体中高度普遍，无论是作为滥用药物还是疼痛管理。它一直 表明HIV/HIV蛋白和滥用药物（例如吗啡）都有助于神经炎症，这是 反过来，涉及炎症的激活。越来越多的证据也表明非编码RNA 作为控制重要细胞功能的表观遗传和转录后调节剂，包括激活， 这改变了各种生物学过程的动态。虽然已经显示了HIV蛋白和阿片类药物 为了改变CNS细胞的基因表达谱，详细的分子存在知识差距 HIV转录剂（TAT）蛋白和 吗啡在星形胶质细胞激活上。该提议的核心假设是艾滋病毒tat和吗啡， 不同的调节机制增强星形胶质细胞激活反过来导致加剧 神经炎症。我们关于SIV-额叶皮质中全基因组硫酸盐测序的初步数据 感染的恒河猕猴显示NLRP6的DNA低甲基化增加（点数样受体家族，pyrin 含结构域的蛋白质6）启动子在NLRP6表达中伴有上调。此外， 人类和小鼠原发性星形胶质细胞暴露于HIV TAT和吗啡也导致下降 MicroRNA-152的表达伴随着NLRP6的表达增加。有趣的是，我们也是如此 发现这些主要星形胶质细胞暴露于HIV TAT或吗啡导致细胞增加 促炎细胞因子（IL1β和IL18）的表达增加，通过凋亡。基于 中央假设和可靠的初步数据，该提案导致了以下特定目的：具体目的 1：确定介导的NLRP6炎性体激活中涉及的分子机制 在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 在HIV TAT和吗啡暴露的星形胶质细胞体外参与NLRP6的启动子DNA降甲基化； 特定目的3：验证HIV TAT和吗啡对NLRP6炎症体介导的组合作用 星形胶质细胞激活，体内。了解负责HIV引起的星形胶质细胞激活的机制 吗啡将为未来发展的新疗法开发奠定基础，旨在使HIV和 优化介导的神经炎症反应。