Early binge drinking and gene regulation

描述（由申请人提供）：INIA的这个新组成部分的总体目标是了解过量饮酒的神经生物学。流行病学数据表明，早期开始饮酒，特别是过度饮酒，是最终酗酒的主要危险因素。我们的假设是，基因表达的先天差异和对酒精的反应差异，特别是在酗酒范式中大量接触酒精的差异，都导致了过度饮酒的发展。我们假设酒精暴露会影响DNA甲基化，这将对基因表达和剪接产生持久影响，从而增加长期过度饮酒的脆弱性。通过在同一组织中以协调的方式检查基因表达和剪接、DNA甲基化和microrna，这三个Specific Aims将紧密联系在一起，这样我们就可以阐明酒精暴露发挥其作用的机制。我们的策略是研究从酒精偏好不同的品系中选择的未接触酒精的动物，找到导致过度饮酒的预先存在的遗传差异，同时研究从出生后30天到58天反复酗酒的动物，以确定这如何影响基因表达和DNA甲基化，以及这种差异是否持续存在。我们将分析暴露和未暴露的动物在重复狂欢结束时的差异，以及30天后的差异，看看哪些差异仍然存在。我们将重点关注INIA的关键区域：伏隔核壳（Nac-shell）、杏仁核中央核（CeA）和腹侧被盖区（VTA）；我们有来自成年男性的微阵列数据，显示这些区域的基因表达存在许多显著差异。将测试在全球研究中发现的差异，以及从人类研究中选择的候选基因，以确定它们是否在cis中受到控制。我们将研究同一组织中的基因表达、剪接、DNA甲基化和microrna，以便对酒精的影响有一个综合的、全系统的了解，这可能是过量饮酒的基础。这些机制研究不能在人类身上进行，因为我们既不能控制酒精暴露，也不能进入脑组织。由于这个原因，一个额外的翻译成分将是比较乙醇对大脑关键区域的影响与可接近的组织，血液中的影响，期待潜在的人体研究。

DESCRIPTION (provided by applicant): The overall aim of this new component for INIA is to understand the neurobiology of excessive alcohol consumption. Epidemiological data demonstrate that early initiation of drinking, and particularly excessive drinking, is a major risk factor for eventual alcoholism. Our hypothesis is that both innate differences in gene expression and differences in response to alcohol, particularity to heavy exposure to alcohol in a binge drinking paradigm, contribute to the development of excessive alcohol drinking. We hypothesize that alcohol exposure will affect DNA methylation which will have lasting effects on gene expression and splicing in a way that increases long-term vulnerability to excessive alcohol drinking. All three Specific Aims will be closely linked, by examining gene expression and splicing, DNA methylation, and microRNAs in a coordinated way on the same tissues, so that we can elucidate mechanisms by which alcohol exposure exerts its effects. Our strategy is to study both alcohol-naive animals from lines selected for difference in alcohol preference, to find pre-existing genetic differences that predispose to excessive drinking, and also to study animals that have experienced repeated binge drinking from postnatal day 30 to 58 to determine how that affects gene expression and DNA methylation, and whether the differences persist. We will analyze differences between exposed and unexposed animals at the end of the repeated binges and also 30 days later to see which differences persist. We will focus on key INIA regions: the shell of the nucleus accumbens (Nac-shell), the central nucleus of the amygdala (CeA), and the ventral tegmental area (VTA); we have microarray data from adult males that show many significant differences in gene expression in these regions. Differences found in global studies, and selected candidate genes from human studies, will be tested to determine whether they are controlled in cis. We will study gene expression, splicing, DNA methylation and microRNAs in the same tissues, to allow an integrated, systems-wide understanding of the effects of alcohol that might underlie excessive alcohol consumption. These mechanistic studies cannot be carried out in humans because we can neither control alcohol exposure nor access brain tissue. For that reason, an additional translational component will be to compare the effects of ethanol on key brain regions with those in an accessible tissue, blood, looking forward to potential human studies. PUBLIC HEALTH RELEVANCE: The proposed studies will provide insight into the mechanisms underlying differences in alcohol preference and excessive alcohol consumption, phenotypes critical in the development of alcoholism. Studying animals exposed to heavy binge drinking should reveal mechanisms that establish increased vulnerability to excessive drinking and alcoholism, and guide the search for better treatments. The studies will complement ongoing work in humans.