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

项目 1 - 调节 DO 小鼠渐进乙醇消耗的新基因网络

• 批准号: 10633317
• 负责人: MICHAEL F MILES
• 金额: $ 18.48万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-05 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10633317
• 关键词: 3-Dimensional; Affect; Alcohol consumption; Alcohol dependence; Alleles; Animals; Behavior; Behavioral; Behavioral Genetics; Bioinformatics; Breeding; CRISPR/Cas technology; Caenorhabditis elegans; Candidate Disease Gene; Chromatin; Collaborations; Complex; Consumption; Data; Detection; Distal; Drosophila genus; Ethanol; Ethanol Metabolism; Funding; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Variation; Genetic study; Genomics; Genotype; Haplotypes; Heavy Drinking; Heritability; Heterozygote; Human; Human Genetics; Human Genome; Injections; Intake; Invertebrates; Investigation; Laboratories; LoxP-flanked allele; Maps; Meta-Analysis; Modeling; Molecular Conformation; Morbidity - disease rate; Mouse Strains; Mus; Nucleus Accumbens; Pathway Analysis; Phenotype; Phylogeny; Play; Prefrontal Cortex; Quantitative Trait Loci; Resources; Rodent; Rodent Model; Role; Scheme; Testing; Therapeutic; Untranslated RNA; Variant; Viral Vector; Water; Work; alcohol behavior; alcohol research; alcohol sensitivity; alcohol use disorder; behavioral genomics; behavioral phenotyping; behavioral study; candidate validation; drinking; experimental study; gene discovery; gene network; genetic analysis; genome wide association study; genome-wide; genomic data; genomic locus; knock-down; mortality; mouse genetics; novel; overexpression; progenitor; screening; trait; transcriptome sequencing; validation studies

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.

项目摘要-项目1 在过去四年的P50资助VCU酒精研究中心（VCU ARC），我们的实验室使用 来自杰克逊实验室（http：//www.example.com）的用于行为遗传学的多样性远交（DO）小鼠，do.jax.org 对渐进式乙醇消耗的初步基因组研究。DO小鼠来源于8个祖细胞小鼠品系 选择最大限度地提高遗传多样性，并利用产生高度杂合性的育种方案， 用于精细绘制复杂的性状，如乙醇消耗。因此，小鼠更忠实地模仿遗传 酒精使用障碍（AUD）。正如预期的那样，基于DO祖细胞中的初步工作， 菌株，使用渐进式乙醇消耗模型（间歇性乙醇获取，IEA）的行为研究 超过600只DO小鼠的摄食量值分布广泛（约0.5-38 g/kg/24 h）， 在4周的实验中，摄入量显著增加。基因分型确定了10个全基因组显著或 LOD ≥ 6且支持区间一般< 2 Mb的暗示性QTL。引人注目的是，数量性状基因座（QTL） 饮酒的第一周与饮酒的最后一周不同。正在进行的单体型分析 和整合来自前额皮质的RNAseq数据已经确定了临时候选基因，包括 其中两个与人类全基因组饮酒相关研究有关， 依赖我们假设在这次更新中，这个DO行为QTL和基因组数据的扩展将 鉴定有助于小鼠乙醇消耗行为的新候选基因和基因网络， 这些数据将为现有和未来的人类基因研究和AUD治疗工作提供信息。 因此，我们的具体目标描述：1）进一步的表达遗传学分析，在细胞核，等位基因特异性 表达分析和染色质3D构象分析，以确定和完善一系列的位置 与初始乙醇摄入量和渐进消费量相关的行为QTL的候选基因; 2） 200只DO小鼠前额叶皮层和丘脑核中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， 3）验证候选基因或网络枢纽在乙醇行为中的功能，包括 使用无脊椎动物模型（与2009年项目2和3合作） 本中心的建议），啮齿动物模型（本项目和啮齿动物行为核心）。此外，这项工作 项目将为项目4和项目5中描述的新的人类基因研究提供信息。在整个 在这个项目中，VCU ARC生物信息学和分析核心将为RNAseq分析提供关键支持 数据和Capture-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的所有其他组成部分产生重要的相互作用 ARC并将告知酒精研究领域，了解过渡到酒精的机制。 滥用性饮酒，以及未来治疗努力潜在靶向的候选基因/机制。