Control of Alcohol Responses by Actin-Regulating Genes

酒精滥用障碍(AUD)是一种主要的健康危害，每年在美国影响数百万人 美国。AUD的风险因素包括对酒精醉人效应的初步抵抗力，以及 对重复暴露的耐受性的发展。AUD也有重要的遗传病因，许多 基因已经被人类全基因组关联研究所牵连，包括我们的研究。 然而，对这些基因中的许多基因的详细分子理解仍然缺乏。“肌动蛋白的调节 细胞骨架“已被大量哺乳动物酒精转录组学研究所牵连，而且主要 肌动蛋白动力学的调节者包括Rho家族的GTP酶及其调节者。我们的发现得到了支持 通过这项竞争性续签拨款的先前资金期限，已经表明Rho GTPase调节器中的SNPs RSU1与酒精依赖和饮酒有关，强调了我们的 学习。我们的总体目标是更好地了解脑出血的分子机制和神经组织特异性。 Rho GTP酶调节器使用果蝇强大的遗传工具包。基于我们之前的发现和 大量的初步数据，我们假设不同的Rho GTP酶调节因子有不同的酒精 反应表型，取决于它们被操纵的神经回路。我们将首先确定 需要Rho家族的rac1 GTPase调节剂的电路和神经递质系统 正常酒精诱导的镇静和耐受。第二，我们将确定酒精镇静和 Rho GTPase信号转导的另外11个成员的耐受性，包括8个与 成瘾/酒精表型在人类GWAS研究中的应用这些候选人是由酒精推荐的- 通过我们的ATAC-SEQ实验确定，它们的DNA调节区的可及性发生了变化。 这些数据还强调了四种转录因子，它们的结合基序在 酒精诱导的可及性DNA调控元件的变化。在第三个目标中，我们将确定 其中两个潜在的主要调节因素是酒精诱导的基因调控和行为改变 酒精反应。使用ATAC-SEQ以组织特异性的方式确定DNA调节元件 突出了我们应用程序的一种主要创新方法。总的来说，这些目标与NIAAA是一致的 重点关注“酒精敏感性和耐受性的遗传学”PA-18-660，其中强调了具体的 感兴趣的研究领域包括：模式生物…中的翻译研究用于体内验证 候选基因…来自人类的研究“，以及”候选基因…的研究“组织特异性基因 表达，以及对生物途径的贡献“。许多以前的出版物都与这笔赠款联系在一起 大量的初步数据证明了调查人员的专业知识和研究的可行性。 因此，我们成熟的从果蝇到人类的翻译方法将直接影响到 了解人类成瘾基因的功能。

Alcohol abuse disorders (AUD) are a major health hazard that affects millions of people every year in the United States. Risk factors for AUD include initial resistance to the intoxicating effects of alcohol, as well as the development of tolerance upon repeat exposure. AUDs also have significant genetic etiology, and many genes have been implicated from human genome-wide association studies (GWAS), including by our studies. A detailed molecular understanding of many of these genes is still lacking, though. “Regulation of the actin cytoskeleton” has been implicated by numerous mammalian alcohol transcriptomics studies, and major regulators of actin dynamics include the Rho family of GTPases, and their regulators. Our findings, supported by prior funding periods of this competitive renewal grant, have shown that SNPs in the Rho GTPase regulator RSU1 are associated with alcohol dependence and drinking, underscoring the translational significance of our studies. Our overall goal is to better understand the molecular mechanisms and neural tissue-specificity of Rho GTPase regulators using the powerful genetic toolkit of Drosophila. Based on our prior findings and extensive preliminary data, we hypothesize that different Rho GTPase regulators have distinct alcohol response phenotypes, depending on the neural circuits they are manipulated in. We will first determine the circuits and neurotransmitter systems that require regulators of the Rac1 GTPase from the Rho family for normal alcohol-induced sedation and tolerance. Second, we will determine the role in alcohol sedation and tolerance of 11 additional members of Rho GTPase signaling, including 8 orthologs of genes associated with addiction/alcohol phenotypes in human GWAS studies. These candidates have been suggested by alcohol- induced changes in accessibility of their DNA regulatory regions, as determined by our ATAC-seq experiments. The data also highlight four transcription factors, based on the overrepresentation of their binding motifs in alcohol-induced changes of accessible DNA regulatory elements. In a third aim, we will determine the role of two of these potential master regulators of alcohol-induced changes in gene regulation and in behavioral alcohol responses. The use of ATAC-seq to determine DNA regulatory elements in a tissue specific manner highlights a major innovative approach of our application. Taken together, these Aims are in line with NIAAA priorities focusing on the “Genetics of Alcohol Sensitivity and Tolerance”PA-18-660, which emphasize specific areas of research interest including “translational research in model organisms…for in vivo validation of candidate genes…from studies in humans”, and “studies of candidate genes…for tissue-specific gene expression, and for contributions to biological pathways”. Many prior publications linked to this grant, together with extensive preliminary data demonstrate the expertise of the investigator and the feasibility of the study. Our proven translational approach from Drosophila to humans will therefore have direct impact on the understanding of the function of human addiction genes.