Role of glucocorticoid receptor-mediated mRNA decay in alcohol dependence

糖皮质激素受体介导的 mRNA 衰减在酒精依赖中的作用

• 批准号: 10811212
• 负责人: Candice Contet
• 金额: $ 25.97万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10811212
• 关键词: Abstinence; Affect; Agonist; Air; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohols; Astrocytes; Attention; Automobile Driving; Binding; Bioinformatics; Brain; Brain region; Chromosome Mapping; Chronic; Code; Complex; Corticosterone; Cultured Cells; Cytoplasm; Dexamethasone; Down-Regulation; Endoribonucleases; Ethanol; Functional disorder; Gene Expression; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; Heavy Drinking; Hyperactivity; Impaired cognition; Inhalation; Ligand Binding; Ligands; Location; Medial; Mediating; Memory; Memory impairment; Mental disorders; Messenger RNA; Mifepristone; Modality; Modeling; Molecular; Motivation; Mus; Play; Prefrontal Cortex; Primates; Process; Proteins; Proteomics; RNA Interference; Receptor Activation; Receptor Inhibition; Receptor Signaling; Recording of previous events; Reproducibility; Research Project Grants; Resources; Rodent; Role; Sampling; Signal Transduction; Specificity; Testing; Therapeutic; Time; Transcript; Up-Regulation; Water; Withdrawal; Work; alcohol abuse therapy; alcohol effect; alcohol exposure; alcohol prevention; alcohol seeking behavior; alcohol use disorder; antagonist; behavior test; cell type; cohort; genetic manipulation; glucocorticoid-induced orphan receptor; human subject; knock-down; mRNA Decay; male; molecular targeted therapies; nervous system disorder; novel; object recognition; pharmacologic; prevent; recruit; sex; small hairpin RNA; transcription factor; transcriptome sequencing; transcriptomics; vapor

SUMMARY Pharmacological inhibition of glucocorticoid receptor (GR) signaling can efficiently reduce alcohol intake and seeking in rodent and primate models of heavy alcohol drinking, as well as in human subjects with an alcohol use disorder (AUD). Despite the wealth of evidence supporting the therapeutic potential of GR inhibition for the treatment of AUD, the molecular mechanism mediating this effect remains unknown. Aside from acting as a transcriptional regulator, GR can bind a subset of mRNAs in the cytoplasm and elicit their rapid degradation upon ligand binding – a process called GR-mediated mRNA decay (GMD). Intriguingly, we demonstrated that the endoribonuclease RIDA, a critical component of the GMD complex, is among the most significantly upregulated proteins in the mouse medial prefrontal cortex (mPFC) during abstinence following a history of excessive alcohol drinking. The present project will test the hypothesis that hyperactive GMD contributes to alcohol intake escalation and underlies the ability of GR antagonism to reduce alcohol drinking in mice withdrawn from chronic intermittent alcohol vapor inhalation. A first aim will be to determine whether abstinence increases GMD activity in excessive alcohol drinkers. To do so, we will first determine the identity of mRNAs bound to GR in mPFC samples from alcohol-naïve mice and test whether GR activation causes their rapid degradation. We will then examine the effect of alcohol withdrawal on these potential GMD substrates and the ability of GR inhibition to prevent it. A second aim will be to determine whether blocking GMD in the mPFC via local RIDA knockdown, which will not impact GR transcriptional activity, can replicate the effect of GR antagonism on excessive alcohol consumption and cognitive impairment. Our approach capitalizes on our expertise in modeling AUD in mice and manipulating gene expression in small brain regions, combined with access to state-of-the-art core resources for RNA sequencing and bioinformatic analysis. The proposed work will enhance our understanding of the molecular mechanisms driving alcohol intake escalation and memory deficits in mice, as well as the mechanism of action of GR antagonists. It will probe for the first time the relevance of GMD in the brain and may identify a new molecular target for the treatment of AUD and other GR-related neurological and psychiatric disorders.

概括 药理学抑制糖皮质激素受体（GR）信号传导可以有效地减少酒精摄入量和 寻求大量饮酒的啮齿动物和灵长类动物模型，以及酒精的人类受试者 使用障碍（AUD）。尽管有大量证据支持GR抑制的治疗潜力 AUD处理，介导这种作用的分子机制仍然未知。除了充当 转录调节剂GR可以结合细胞质中mRNA的子集并引起其快速降解 配体结合 - 一种称为GR介导的mRNA衰变（GMD）的过程。有趣的是，我们证明了 GMD复合物的关键组成部分的内核酸酶RIDA是最重要的 在节制期间，小鼠培养基前额叶皮层（MPFC）中上调蛋白质 过量饮酒。本项目将测试过度活跃的GMD有助于的假设 由于GR拮抗的能力 从慢性间歇性酒精蒸气吸入。第一个目的是确定禁欲是否增加 GMD活动过多的酒精饮用者。为此，我们将首先确定与GR结合的mRNA的身份 在未经酒精的小鼠的MPFC样品中，测试GR激活是否会导致它们的快速降解。我们 然后，将检查戒酒对这些潜在GMD底物的影响和GR的能力 抑制以防止它。第二个目的是确定是否通过本地RIDA阻止MPFC中的GMD 敲低不会影响GR转录活动，可以复制GR拮抗对 过度饮酒和认知障碍。我们的方法利用了我们在建模方面的专业知识 小鼠中的aud和在小脑区域操纵基因表达，并加入最新 RNA测序和生物信息学分析的核心资源。拟议的工作将增强我们的 了解驱动酒精摄入量升级的分子机制，并在小鼠中定义记忆 以及GR拮抗剂的作用机理。它将首次探测GMD在 大脑，可能会确定一个新的分子靶标，用于治疗AUD和其他与GR相关的神经系统 精神疾病。