Department of Cell and Molecular Pharmacology & Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USA; Department of Psychiatry, Stellenbosch University, Cape Town, South Africa; South African Medical Research Council / Stellenbosch University Genomics of Brain Disorders Research Unit, Department of Psychiatry, Stellenbosch University, Cape Town, South Africa.
Department of Cell and Molecular Pharmacology & Experimental Therapeutics, Medical University of South Carolina, Charleston, SC 29425, USA.
Biomed Pharmacother. 2024 Nov;180:117514. doi: 10.1016/j.biopha.2024.117514. Epub 2024 Oct 2.
Alcohol use disorder (AUD) is the most prevalent substance use disorder but there is incomplete knowledge of the underlying molecular etiology. Here, we examined the cytosolic proteome from the nucleus accumbens core (NAcC) of ethanol drinking rhesus macaques to identify ethanol-sensitive signaling proteins. The targets were subsequently investigated using bioinformatics, genetic, and pharmacological manipulations in mouse models of ethanol drinking. Of the 1000+ cytosolic proteins identified in our screen, 50 proteins differed significantly between control and ethanol drinking macaques. Gene Ontology analysis of the differentially expressed proteins identified enrichment in pathways regulating metabolic processes and proteasome activity. Because the family of Glutathione S-transferases (GSTs) was enriched in these pathways, validation studies targeted GSTs using bioinformatics and genetically diverse mouse models. Gstp1 and Gstm2 were identified in Quantitative Trait Loci and published gene sets for ethanol-related phenotypes (e.g., ethanol preference, conditioned taste aversion, differential expression), and recombinant inbred strains that inherited the C57BL/6J allele at the Gstp2 interval consumed higher amounts of ethanol than those that inherited the DBA/2J allele. Genetic deletion of Gstp1/2 led to increased ethanol consumption without altering ethanol metabolism or sucrose preference. Administration of the pharmacologic activator of Gstp1/2, carnosic acid, decreased voluntary ethanol drinking. Proteomic analysis of the NAcC cytosolic of heavy drinking macaques that were validated in mouse models indicate a role for glutathione-mediated redox regulation in ethanol-related neurobiology and the potential of pharmacological interventions targeting this system to modify excessive ethanol drinking.
酒精使用障碍(AUD)是最常见的物质使用障碍,但对其潜在的分子病因知之甚少。在这里,我们检查了酒精饮用恒河猴伏隔核核心(NAcC)的细胞质蛋白质组,以鉴定乙醇敏感的信号蛋白。随后使用生物信息学、遗传和药理学方法在酒精饮用小鼠模型中对这些靶点进行了研究。在我们的筛选中鉴定出的 1000 多种细胞质蛋白中,有 50 种蛋白在对照组和酒精饮用猕猴之间存在显著差异。差异表达蛋白的基因本体分析确定了代谢过程和蛋白酶体活性调节途径的富集。由于谷胱甘肽 S-转移酶(GSTs)家族在这些途径中富集,因此使用生物信息学和遗传多样化的小鼠模型对 GSTs 进行了验证研究。Gstp1 和 Gstm2 被鉴定为与乙醇相关表型(例如,乙醇偏好、条件性味觉厌恶、差异表达)的数量性状基因座和已发表的基因集,以及遗传了 C57BL/6J 等位基因的重组近交系在 Gstp2 区间的乙醇消耗量高于遗传了 DBA/2J 等位基因的重组近交系。Gstp1/2 的基因缺失导致乙醇消耗增加,而不改变乙醇代谢或蔗糖偏好。Gstp1/2 的药理激活剂 carnosic acid 的给药减少了自愿性乙醇摄入。在小鼠模型中验证的重度饮酒猕猴的伏隔核细胞质的蛋白质组学分析表明,谷胱甘肽介导的氧化还原调节在乙醇相关神经生物学中起作用,以及针对该系统的药理学干预改变过度饮酒的潜力。