Project 1 - Novel gene networks modulating progressive ethanol consumption in DO mice

项目摘要-项目1 在过去四年P50资助VCU酒精研究中心(VCU ARC)期间，我们的实验室使用 来自杰克逊实验室的多样性杂交(DO)小鼠(http://do.jax.org)用于行为遗传学和 渐进性酒精消费的初步基因组研究。小鼠是否来自8个祖代小鼠品系 选择使遗传多样性最大化并利用产生高度杂合性的育种方案 用于精细定位复杂的特征，如酒精消耗量。因此，老鼠是否更忠实地模仿基因 酒精使用障碍(AUD)的方面。正如预期的那样，基于DO前体的初步工作 菌株，使用渐进式乙醇消耗模型的行为研究(间歇性乙醇获取，IEA) 在600多只DO小鼠身上，显示出广泛的消耗值分布(~0.5-38g/kg/24小时)，转变为 在四周的实验中显著增加了摄入量。基因分型确定了10个全基因组显著或 建议的≥带有LOD QTL6和支持间隔，一般为&lt；2Mb。值得注意的是，数量性状基因座(QTL) 饮酒的第一周不同于最后一周的饮酒。正在进行的单倍型分析 和整合来自前额叶皮质的RNAseq数据已经确定了临时候选基因，包括 两个与饮酒或饮酒有关的人类全基因组关联研究 依赖。我们假设在这次更新中，行为QTL和基因组数据的扩展将 识别新的候选基因和基因网络，这些基因和网络有助于小鼠和 这些数据将为现有和未来的人类基因研究和AUD治疗努力提供信息。 因此，我们的具体目标描述如下：1)进一步的伏隔核表达遗传学分析，等位基因特异性 表达分析和染色质3D构象分析，以识别和提炼位置列表 行为QTL候选基因与初始酒精摄入量与渐进饮酒量相关；2) 200只DO小鼠前额叶皮质和伏隔核RNAseq数据的基因网络分析 确定与消费密切相关的网络和可能的机制 上周；以及3)候选基因或网络枢纽在酒精行为中发挥作用的验证，包括 使用无脊椎动物模型初步或逐步消耗乙醇(与项目2和项目3合作 该中心提案)、啮齿动物模型(该项目和啮齿动物行为核心)。此外，这个机构的工作 项目将向项目4和项目5中描述的新的人类基因研究提供信息，并由项目5提供信息。 该项目，VCU ARC生物信息学和分析核心将为RNAseq的分析提供关键支持 数据和捕获-C染色质构象研究，以及候选基因和网络的选择。这 新的基因发现和网络分析工作将与VCU的所有其他组件产生重大相互作用 并将向酒精研究领域提供信息，了解向 滥用饮酒，以及未来治疗努力潜在靶向的候选基因/机制。

Project Summary – Project 1 Over the last four years of P50 funding to the VCU Alcohol Research Center (VCU ARC), our laboratory used Diversity Outbred (DO) mouse mice from Jackson Laboratories (http://do.jax.org) for behavioral genetics and initial genomic studies on progressive ethanol consumption. DO mice originate from 8 progenitor mouse strains chosen to maximize genetic diversity and utilize a breeding scheme producing a high degree of heterozygosity for fine mapping complex traits such as ethanol consumption. As such, DO mice more faithfully mimic genetic aspects seen with alcohol use disorder (AUD). As expected, based upon preliminary work in DO progenitor strains, behavioral studies using a progressive ethanol consumption model (intermittent ethanol access, IEA) on over 600 DO mice showed a broad distribution of consumption values (~0.5–38 g/kg/24h) that shifted to significantly higher intake over the 4 week experiment. Genotyping identified 10 genome-wide significant or suggestive QTL with LOD ≥ 6 and support intervals generally < 2 Mb. Strikingly, quantitative trait loci (QTL) for the first week of drinking differed from those during the last week of consumption. Ongoing haplotype analysis and integration of RNAseq data from prefrontal cortex have identified provisional candidate genes, including two that have been implicated in human genome-wide association studies on alcohol consumption or dependence. We hypothesize in this renewal that extension of this DO behavioral QTL and genomic data will identify novel candidate genes and gene networks contributing to ethanol consumption behaviors in mice and that such data will inform existing and future human genetic studies and therapeutic efforts on AUD. Our specific aims thus describe: 1) Further expression genetics analysis in nucleus accumbens, allele specific expression analysis, and chromatin 3D conformation analyses to identify and refine a list of positional candidate genes for behavioral QTL associated with initial ethanol intake vs. progressive consumption; 2) Gene network analysis of RNAseq data in both prefrontal cortex and nucleus accumbens across 200 DO mice to identify networks and possible mechanisms tightly associated with consumption differing between first and last week; and 3) Validation of candidate genes or network hubs as functioning in ethanol behaviors, including initial or progressive ethanol consumption, using invertebrate models (collaboration with Projects 2 and 3 of this Center proposal), rodent models (this project and Rodent Behavioral Core). Further, the work of this project will inform and be informed by novel human genetic studies described in Projects 4 and 5. Throughout this project, the VCU ARC Bioinformatics and Analysis Core will provide critical support for analysis of RNAseq data and Capture-C chromatin conformation studies, and the choice of candidate genes and networks. This novel gene discovery and network analysis effort will have major interactions with all other components of VCU ARC and will inform the field of alcohol research with understanding of mechanisms involved in the transition to abusive drinking, and candidate genes/mechanisms for potential targeting by future therapeutic efforts.