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: 10473784
• 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/10473784
• 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-catenin的miRNA调节触发BK通道酒精耐受性和 准备的消费 研究已将大电导电压电压和钙激活通道（BK）视为 神经元刺激性的关键调节剂，与行为酒精耐受性有关 无脊椎动物。同样，在哺乳动物中，通过淘汰降低BK通道酒精敏感性 它的ß4亚基导致小鼠的酒精自愿消耗增加。对乙醇的敏感性 分子水平的特征是通道活性的急性增强。但是，不同 BK通道的同工型显示酒精敏感性的变化，分布不同 响应长时间的暴露，在质膜表面上，也称为持续 酒精耐受性（PMT）。对酒精敏感的同工型的miRNA靶向和活性 据信，BK渠道的内在化是建立乙醇的关键 稳态适应会导致纹状体内持续变化。这些 机制可以独立运行，同一终点的增加 酒精不敏感的BK通道的代表或互动以编排和优化这些 在调节的翻译和贩运水平上，持续的适应性变化。 我们当前项目的结果已将Wnt/β-catenin信号通路确定为钥匙 BK通道表面重新分布的调节剂，以响应6hr乙醇暴露。我们有 将其描述为长期神经元可塑性的一种形式，需要蛋白质合成 导致β-catenin的表达和持久性增加，超过24小时。激活 ß-catenin信号传导与人类癌细胞中的miR-9上调有关，以及 抑郁症小鼠中的miRNA表达。值得注意的是，miR-9在 纹状体是PMT介导BK通道的急性ETOH脱敏的关键事件。那， 该提议的总体假设是一个单一的“暴饮暴食” 6小时暴露 通过miRNA调节的Wnt/β-catenin在NAC内变化BK通道表面分布 信号，影响随后的饮酒。我们将使用电生理学，分子 生物学，行为测定和成像技术，以确定Wnt/ß-catenin的作用 纹状体内BK贩运和miRNA调节的信号传导，目的是 识别涉及的新型热目标，以防止最初的过渡到 强迫性饮酒行为。