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）对退伍军人的心理和身体健康有实质性的负面影响， 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 美国。每天增加阿片类药物剂量增加的死亡风险增加了 镇痛作用。退伍军人卫生管理局（VHA）认识到成功的临床挑战 为我们的退伍军人安全开处方阿片类药物。吗啡提取（MW）是鸦片滥用的决定者之一 在Oud个人中，其机制知之甚少，并且仍然缺乏有效的疗法。 MW激活星形胶质细胞以释放神经炎性因子。新兴证据表明 反应性星形胶质细胞的神经炎性因子Lipocalin 2（LCN2）与多种神经有关 系统损伤和神经炎症。我们的初步数据表明，赞助的MW（S-MW）诱导 来自星形胶质细胞的TLR4，NF-KB（RERA/P65）和LCN2。 LCN2受体（LCN2-R）位于脑干 S-MW中的灰灰色（PAG）神经元。表观遗传作者EZH2通过产生沉默的基因表达 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，并改善了PAG中的抗氧化线粒体Sirtuin 3（SIRT3）。确切的 S-MW中星形胶质细胞对神经元活性的分子机制知之甚少。在提案中， 我们将检验以下假设，即神经胶质活性诱导星形胶质细胞衍生的LCN2的释放，该假设降低了。 神经元抗氧化SIRT3并最终增加了S-MW小鼠的PAG中的PCREB。特定目标1： 确定星形胶质细胞激活是否在TLR4和NF-κB中诱导LCN2的过表达 带有S-MW的小鼠的VLPAG。特定目的2：定义LCN2-R是否介导降低的SIRT3表达式 导致S-MW小鼠VLPAG神经元的PCREB增加。我们工作的关键特征是 能够偶尔使用新的遗传/表观遗传和分子测定，细胞类型特异性敲除 （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。 提案将为阿片类药物戒断的发病机理提供重要的见解，并阐明了一种新颖 阿片类药物戒断阿片类药物的治疗靶标。这项研究将来会导致 基于我们确定，为普通人群和退伍军人开发新药 TLR4的作用机理--- LCN2 --- SIRT3 --- pCREB途中撤离期间的PCREB途径。