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

基本信息

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
项目类别：
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

项目摘要

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，但大脑仍继续成为艾滋病毒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药物滥用的共存病。与艾滋病毒相关的神经认知障碍和神经艾滋病毒在多种滥用药物中，阿片类药物的使用最为严重。它在艾滋病毒感染者中非常普遍，无论是作为滥用药物还是用于疼痛管理。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，越来越多的证据也表明，非编码 RNA 会被激活。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，而艾滋病毒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，进而导致加剧我们对 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。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。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焦亡激活促炎细胞因子（IL1β和IL18）的表达增加。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的体外实验；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了解体内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。