MiR-9 regulation of beta-catenin mediated alcohol tolerance and EtOH consumption

MiR-9 对 β-连环蛋白介导的酒精耐受性和 EtOH 消耗的调节

• 批准号: 10262926
• 负责人: CRISTINA M. VELAZQUEZ
• 金额: $ 33.75万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10262926
• 关键词: Acute; Adolescent; Alcohol abuse; Alcohol consumption; Alcoholism; Alcohols; Behavioral; Behavioral Assay; Brain region; Calcium; Cell Nucleus; Cell membrane; Chemosensitization; Consumption; Corpus striatum structure; Coupled; Cytoplasm; Data; Development; Disease; Dissociation; Electrophysiology (science); Elements; Ethanol; Event; Goals; Hour; Human; Image; Imaging Techniques; Invertebrates; Ion Channel; Knock-out; Knockout Mice; Knowledge; Learning; Link; Mammals; Measures; Mediating; Membrane; Memory; Mental Depression; Messenger RNA; MicroRNAs; Modeling; Molecular; Molecular Biology; Monitor; Mus; Names; Neuronal Plasticity; Neurons; Nucleus Accumbens; Pathway interactions; Play; Preparation; Process; Protein Biosynthesis; Protein Isoforms; Publishing; Regulation; Research; Role; Signal Pathway; Signal Transduction; Slice; Surface; Synaptic plasticity; System; Testing; Transcriptional Activation; Transgenic Mice; Up-Regulation; Variant; Withdrawal; addiction; alcohol exposure; alcohol response; alcohol sensitivity; beta catenin; binge drinking; cancer cell; comparative; desensitization; drinking behavior; hippocampal pyramidal neuron; in vitro Model; in vivo; large-conductance calcium-activated potassium channels; neuronal excitability; new therapeutic target; overexpression; preference; prevent; response; trafficking; voltage

PROJECT SUMMARY miRNA regulation of Wnt/beta-catenin triggers BK channel alcohol tolerance and facilitated consumption Research has identified the large conductance voltage- and calcium-activated channel (BK) as a key regulator of neuronal excitability genetically associated to behavioral alcohol tolerance in invertebrates. Similarly in mammals, decreasing BK channel alcohol sensitivity by knocking out its ß4 subunit leads to increased alcohol voluntary consumption in mice. Sensitivity to ethanol at the molecular level is characterized by acute potentiation of channel activity. However, different isoforms of the BK channel show variations in alcohol sensitivity and are differentially distributed on the plasma membrane surface in response to prolonged exposure, also known as persistent alcohol tolerance (PMT). miRNA targeting of alcohol-sensitive isoforms coupled with active internalization of BK channels in response to ethanol are believed to be key in establishing homeostatic adaptations that produce persistent changes within the striatum. These mechanisms may operate independently, with the same end point of increasing the representation of alcohol insensitive BK channels or interact to orchestrate and optimize these persistent adaptive changes at the translational and trafficking level of regulation. Results from our current project have identified the Wnt/β-catenin signaling pathway as a key regulator of BK channel surface redistribution in response to 6hr ethanol exposure. We have characterized this as a form of long-term neuronal plasticity, which requires protein synthesis resulting in increased ß-catenin expression and persistence, past 24hr withdrawal. Activation of ß-catenin signaling has been linked to miR-9 upregulation in human cancer cells, as well as, miRNA expression in depression-resilient mice. Notably, miR-9 upregulated expression in the striatum is a key event in PMT mediating acute EtOH desensitization of BK channels. Thus, the overarching hypothesis of this proposal is that a single “binge-like” 6hr exposure persistently alters BK channel surface distribution within the NAc via miRNA regulated Wnt/β-catenin signaling, impacting subsequent alcohol consumption. We will use electrophysiology, molecular biology, behavioral assays and imaging techniques to determine the role of Wnt/ß-catenin signaling in both BK trafficking and miRNA regulation within the striatum with the goal of identifying novel therapeutic targets involved in preventing the transition from initial to compulsive drinking behavior.

项目摘要 Wnt/β-连环蛋白的miRNA调节触发BK通道酒精耐受和 便利消费 研究已经确定大电导电压和钙激活通道（BK）为 神经元兴奋性的一个关键调节因子，在遗传上与行为酒精耐受性相关， 无脊椎动物类似地，在哺乳动物中，通过敲除 其β 4亚基导致小鼠酒精自愿消耗增加。乙醇敏感性， 分子水平的特征在于通道活性的急性增强。但不同 BK通道的同种型显示出酒精敏感性的变化，并且差异分布 在质膜表面的反应，长期暴露，也称为持久性 酒精耐受性（PMT）酒精敏感亚型的miRNA靶向与活性偶联 BK通道对乙醇应答的内化被认为是建立 在纹状体内产生持续变化的自我平衡适应。这些 这些机制可以独立地操作，具有相同的终点， 代表酒精不敏感的BK通道或相互作用，以协调和优化这些 持续的适应性变化在翻译和运输水平的监管。 我们目前项目的结果已经确定Wnt/β-catenin信号通路是一个关键的 BK通道表面再分布的调节剂对6小时乙醇暴露的响应。我们有 将其描述为一种需要蛋白质合成的长期神经元可塑性 导致24小时停药后β-连环蛋白表达和持续性增加。激活 β-连环蛋白信号传导与人类癌细胞中的miR-9上调有关，此外， 抗抑郁小鼠中的miRNA表达。值得注意的是，miR-9上调表达在 纹状体是PMT介导BK通道急性EtOH脱敏的关键事件。因此 这一建议的首要假设是，一个单一的“暴食式”6小时暴露持续 通过miRNA调节Wnt/β-catenin改变NAc内BK通道表面分布 信号，影响随后的酒精消费。我们将使用电生理学，分子 生物学、行为分析和成像技术来确定Wnt/β-连环蛋白的作用 纹状体内BK运输和miRNA调节的信号传导，目的是 确定新的治疗靶点， 强迫性饮酒行为