The molecular mechanisms of astrocytes-neurons interaction in the morphine use disorder

吗啡使用障碍中星形胶质细胞-神经元相互作用的分子机制

• 批准号: 10487821
• 负责人: SHUANGLIN HAO
• 金额: --
• 依托单位: MIAMI VA HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2026-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487821
• 关键词: Analgesics; Astrocytes; Behavior; Biological Assay; Biological Markers; Brain Stem; Chronic; Chronic Disease; Clinical; Data; Development; Disease; Dose; EZH2 gene; Economics; Epidemic; Epigenetic Process; Future; Gene Expression; Gene Silencing; Gene Transfer; General Population; Genetic; Genetic Transcription; Goals; Housing; Individual; Knockout Mice; LCN2 gene; Lead; Mediating; Mental Health; Methylation; Mission; Mitochondria; Molecular; Morphine; Mus; NF-kappa B; Nervous System Trauma; Neuroglia; Neurons; Opiate Addiction; Opioid; Pathogenesis; Pathway interactions; Patient Care; Patients; Performance at work; Pharmaceutical Preparations; Physical Dependence; Pilot Projects; Psychological Dependence; Recurrent disease; Regulation; Relapse; Role; Sirtuins; Social Functioning; Substance Withdrawal Syndrome; TLR4 gene; Testing; Therapeutic Intervention; United States; United States Department of Veterans Affairs; Veterans; Veterans Health Administration; Viral; Withdrawal; Work; addiction; cell type; combat; conditional knockout; economic cost; effective therapy; insight; midbrain central gray substance; mortality risk; neuroinflammation; new therapeutic target; novel therapeutics; opioid abuse; opioid epidemic; opioid use; opioid use disorder; opioid withdrawal; overdose death; overdose risk; overexpression; p65; pharmacologic; physical conditioning; prescription opioid; promoter; receptor; transcription factor

Opioid use disorder (OUD) has substantial negative consequences on veterans' mental and physical health, work performance, housing status, and social function. Opioid drug abuse has reached an epidemic level in the United States. There is an increased risk of overdose death with higher daily opioid doses for increasing analgesic effect. The Veterans Health Administration (VHA) recognizes the clinical challenges to successfully prescribing opioids safely for our veterans. Morphine withdrawal (MW) is one of determinants of opiate abuse in OUD individuals, yet its mechanisms are poorly understood, and effective therapies are still lacking. MW activates astrocytes to release neuroinflammatory factors. Emerging evidence shows that neuroinflammatory factor Lipocalin 2 (LCN2) from reactive astrocytes is associated with a variety of nervous system injuries and neuroinflammation. Our preliminary data showed that spontaneous MW (S-MW) induced TLR4, NF-kB (RelA/p65), and LCN2 from astrocytes. LCN2 receptor (LCN2-R) is located in the brainstem periaqueductal gray (PAG) neurons in S-MW. Epigenetic writer EZH2 silences gene expression by generating a methylated epigenetic mark at H3K27me3. Our preliminary data showed that S-MW increased neuronal EZH2 and pCREB, and decreased anti-oxidative mitochondrial sirtuin 3 (Sirt3) in the PAG. The exact molecular mechanisms of astrocytes to neuron activity in S-MW remain poorly understood. In the proposal, we will test the hypothesis that glial activity induces the release of astrocytes-derived LCN2, which lowers neuronal anti-oxidative Sirt3 and finally increases pCREB in the PAG in mice with S-MW. Specific Aim 1: To determine whether astrocytic activation induces the overexpression of LCN2 through TLR4 and NF-κB in the vlPAG in mice with S-MW. Specific Aim 2: To define whether LCN2-R mediates the lowered Sirt3 expression leading to pCREB increases in the vlPAG neurons in mice with S-MW. A crucial feature of our work is the ability to causally use new genetic/epigenetic and molecular assay, cell-type specific conditional knockout (cKO) mice, cell type-selective viral-mediated gene transfer, and molecular-pharmacological approach. The proposal will provide important insights into the pathogenesis of opioid withdrawal, and shed light on a novel therapeutic target for opioid withdrawal of opioid withdrawal. The study will could lead, in the future, to the development of new drugs for both the general population and veterans, based on our identification of the mechanism of action of TLR4---LCN2---Sirt3---pCREB pathway during opioid withdrawal.

阿片类药物使用障碍（OUD）对退伍军人的身心健康产生严重的负面影响， 工作表现、住房状况和社会功能。阿片类药物滥用已达到流行病水平 美国。随着每日阿片类药物剂量的增加，过量死亡的风险也会增加 镇痛作用。退伍军人健康管理局 (VHA) 认识到成功实施临床治疗所面临的挑战 为我们的退伍军人安全地开阿片类药物。吗啡戒断（MW）是阿片类药物滥用的决定因素之一 在 OUD 个体中，但对其机制知之甚少，并且仍然缺乏有效的治疗方法。 MW 激活星形胶质细胞释放神经炎症因子。新出现的证据表明 来自反应性星形胶质细胞的神经炎症因子脂质运载蛋白 2 (LCN2) 与多种神经 系统损伤和神经炎症。我们的初步数据表明自发MW（S-MW）诱导 来自星形胶质细胞的 TLR4、NF-kB (RelA/p65) 和 LCN2。 LCN2受体（LCN2-R）位于脑干 S-MW 中的导水管周围灰质 (PAG) 神经元。表观遗传作家 EZH2 通过生成 H3K27me3 处的甲基化表观遗传标记。我们的初步数据表明，S-MW 增加了神经元 EZH2 和 pCREB，并减少 PAG 中的抗氧化线粒体 Sirtuin 3 (Sirt3)。确切的 星形胶质细胞对 S-MW 神经元活动的分子机制仍知之甚少。在提案中， 我们将检验以下假设：神经胶质细胞活性会诱导星形胶质细胞衍生的 LCN2 的释放，从而降低 神经元抗氧化 Sirt3 并最终增加 S-MW 小鼠 PAG 中的 pCREB。具体目标 1： 确定星形胶质细胞活化是否通过 TLR4 和 NF-κB 诱导 LCN2 过度表达 S-MW 小鼠中的 vlPAG。具体目标 2：确定 LCN2-R 是否介导 Sirt3 表达降低 导致 S-MW 小鼠 vlPAG 神经元中的 pCREB ​​增加。我们工作的一个重要特点是 能够因果地使用新的遗传/表观遗传和分子测定、细胞类型特异性条件敲除 (cKO) 小鼠、细胞类型选择性病毒介导的基因转移和分子药理学方法。这 该提案将为阿片类药物戒断的发病机制提供重要见解，并揭示一种新的方法 阿片类药物戒断的治疗目标阿片类药物戒断。这项研究将来可能会导致 根据我们对药物的鉴定，为普通大众和退伍军人开发新药 阿片类药物戒断过程中TLR4---LCN2---Sirt3---pCREB通路的作用机制