Molecular and cellular mechanisms of novel targets in alcohol reward

基因表达的持续变化可能介导许多酒精效应，包括奖励学习， 耐受性和依赖性，这表明有效改变酒精诱导的基因表达的药物应被视为治疗剂。通过抑制调节染色质结构的酶靶向基因表达的药物（表观遗传药物）已广泛用于癌症研究，最近成为神经退行性疾病和药物成瘾的潜在治疗药物。本研究的主要目的是：1）通过检测表观遗传药物对酒精消耗和条件性位置偏爱（CPP）的影响，确定影响酒精奖赏的表观遗传药物; 2）研究表观遗传药物对奖赏通路中包括腹侧被盖区（VTA）和丘脑核（NA）在内的基因表达和细胞生理的影响。总的假设是，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学来降低酒精的奖励特性。为了解决这个研究问题，我们将使用药理学，行为学，基因组学和电生理学方法的组合。酒精将通过使用酗酒小鼠模型的自愿消费或通过全身注射来产生CPP来递送。表观遗传药物将在酒精之前通过全身注射递送。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整体基因表达。腹侧被盖区多巴胺神经元将使用与基因表达平行的电生理学技术进行测试。将使用qRT-PCR和表观遗传学试验验证微阵列结果。电生理学和基因表达数据的整合将阐明药物的作用机制，并确定药物开发的新靶点。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.