Molecular and cellular mechanisms of novel targets in alcohol reward

基因表达的持续变化可能会介导许多酒精效应，包括奖赏学习， 耐受性和依赖性，提示有效改变酒精诱导的基因表达的药物应被视为治疗药物。通过抑制调节染色质结构的酶来靶向基因表达的药物(表观遗传药物)已广泛应用于癌症研究，最近成为治疗神经退行性疾病和药物成瘾的潜在药物。本项目的主要目标是：1)通过测试表观遗传药物对酒精消费和条件性位置偏爱的影响，寻找影响酒精奖赏的表观遗传药物；2)研究所选表观遗传药物对酒精奖赏通路中腹侧被盖区(VTA)和伏隔核(NA)基因表达和细胞生理的影响。总体假设是，一些表观遗传药物将通过改变奖励途径中的基因表达和细胞生理来降低酒精的奖励特性。为了解决这一研究问题，我们将结合药理学、行为学、基因组学和电生理学方法。酒精将通过自愿饮用、使用狂饮的小鼠模型或通过全身注射产生CPP来提供。表观遗传药物将在酒精之前通过全身注射给予。我们将使用激光捕获显微解剖来解剖VTA和NA。我们将在表观遗传药物、载体和酒精的不同组合后测量这些区域的全球基因表达。VTA多巴胺神经元将在基因表达的同时使用电生理技术进行测试。微阵列结果将使用qRT-PCR和表观遗传学分析进行验证。电生理和基因表达数据的结合将阐明药物的作用机制，并为药物开发确定新的靶点。这项研究将为表观遗传药物治疗酒精成瘾的治疗潜力提供初步的机制证据。

Persistent changes in gene expression may mediate many alcohol effects including reward learning, tolerance and dependence, suggesting that agents effective at changing alcohol-induced gene expression should be considered as therapeutic agents. Drugs targeting gene expression through inhibition of enzymes that regulate chromatin structure (epigenetic drugs) have been widely used in cancer research and recently emerged as potential therapeutics for neurodegenerative disorders and drug addiction. The main goals of this project are: 1) to identify epigenetic drugs that affect alcohol reward through testing their effects on alcohol consumption and conditioned place preference (CPP) and 2) to investigate the effects of selected epigenetic drugs on gene expression and cellular physiology in the reward pathway including ventral tegmental area (VTA) and the nucleus accumbens (NA). The overall hypothesis is that some epigenetic drugs will reduce the rewarding properties of alcohol through changes in gene expression and cellular physiology in the reward pathway. To address this research problem, we will use a combination of pharmacological, behavioral, genomic and electrophysiological approaches. Alcohol will be delivered via voluntary consumption using a mouse model of binge drinking or via systemic injections to produce CPP. Epigenetic drugs will be delivered via systemic injections prior to alcohol. We will use laser capture microdissections to dissect VTA and NA. We will measure global gene expression in these regions after different combinations of epigenetic drugs, vehicle and alcohol. VTA dopamine neurons will be tested using electrophysiological techniques in parallel with gene expression. Microarray results will be validated using qRT-PCR and epigenetic assays. Integration of electrophysiological and gene expression data will elucidate the drug's mechanisms of action and identify novel targets for drug development. This research will provide initial mechanistic evidence for the therapeutic potential of epigenetic drugs in treating alcohol addiction.