Functional genomics of ethanol craving and naltrexone

描述（由申请人提供）：尽管我们已经了解了乙醇作用的分子位点，但仍然很少有有效的治疗酒精中毒的方法。纳曲酮（NTX）可减少酗酒者的再犯和乙醇消耗量。NTX是一种非选择性阿片拮抗剂，在动物模型中也被证明可以减少乙醇饮酒行为，包括在复发饮酒模型中阻断增加的乙醇饮酒，即乙醇剥夺效应（EDE）。这些反应的分子机制尚不完全清楚。我们假设，通过研究与纳曲酮作用、EDE和纳曲酮对EDE的影响相关的全基因组基因表达模式，我们可能会对与复发饮酒行为相关的机制有新的认识。在本项目中，高密度寡核苷酸阵列将首先用于表征NTX在幼年C57BL/6小鼠中引起的基因表达模式。研究C57BL/6小鼠的腹侧被盖区、伏隔核和内侧前额叶皮层脑区。NTX的表达谱也将与另外两种减少乙醇饮用或EDE的药物——阿坎前列腺素和MPEP（一种mGluR5谷氨酸受体抑制剂）的表达谱进行比较。目标二将使用阵列研究NTX在乙醇自我给药的两瓶选择模型中对乙醇剥夺引起的基因表达模式的作用。通过对目标1-2中与NTX作用相关的联合表达模式的数据挖掘，我们将根据其表达变化的细胞模式来描述特定的候选基因。在目标3中，候选基因将在两瓶选择模型中评估其在乙醇饮用或EDE中的作用。药理学或遗传学（病毒载体，反义寡核苷酸）手段将被用来改变候选基因的表达在行为测试之前。这些研究应该为EDE的机制和NTX在改变乙醇饮用行为中的作用机制提供新的见解。总之，这些发现可能为酒精中毒的治疗干预确定新的靶点。

DESCRIPTION (provided by applicant): Although much has been learned about molecular sites of action for ethanol, there remains few effective treatments for alcoholism. Naltrexone (NTX) reduces recidivism and ethanol consumption in alcoholics. NTX, an un-selective opioid antagonist, has also been shown to decrease ethanol drinking behavior in animal models, including blocking increased ethanol drinking in a model of relapse drinking, the ethanol deprivation effect (EDE). The molecular mechanism(s) for these responses are not entirely understood. We hypothesize that by studying genome-wide gene expression patterns associated with naltrexone action, EDE and naltrexone effects on EDE, we might gain novel insight into mechanisms relevant to relapse drinking behavior. In this project high-density oligonucleotide arrays will first be used to characterize gene expression patterns evoked by NTX in naive C57BL/6 mice. Ventral tegmental area, nucleus accumbens and medial prefrontal cortex brain regions in C57BL/6 mice will be studied. Expression profiles of NTX will also be compared to those from two other agents that decrease ethanol drinking or the EDE, acamproste and MPEP, an inhibitor of the mGluR5 glutamate receptor. Aim two will then use arrays to study action of NTX on gene expression patterns evoked by ethanol-deprivation in a 2-bottle choice model of ethanol self administration. Through data mining the combined expression patterns related to NTX action in aims 1-2, we will then characterize particular candidate genes in regard to cellular patterns of their expression changes. In Aim 3, candidate genes will be evaluated for their role in ethanol drinking or the EDE in a 2-bottle choice model. Pharmacological or genetic (viral vectors, antisense oligonucleotides) means will be used to alter the expression of candidate genes prior to behavioral testing. These studies should provide novel insight into the mechanisms of the EDE and mechanisms of NTX action in altering ethanol drinking behavior. Together, these findings may identify novel targets for therapeutic intervention in alcoholism.