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资助向VCU酒精研究中心（VCU ARC）的资金中，我们的实验室使用了 杰克逊实验室（http://do.jax.org）的多样性杂种（DO）小鼠行为遗传学和 进行性乙醇消耗的最初基因组研究。小鼠起源于8个祖细胞菌株 选择最大化遗传多样性并利用产生高度杂合性的繁殖方案 用于精细的映射复杂性状，例如乙醇消耗。因此，小鼠会更忠实地模仿通用 酒精使用障碍（AUD）看到的方面。不出所料，基于祖先的初步工作 菌株，使用进行性乙醇消耗模型（间歇性乙醇访问，IEA）的行为研究 在超过600多只小鼠上，消费值（〜0.5-38 g/kg/24h）的分布广泛，该值移至 在4周的实验中，摄入量明显更高。基因分型确定了10个基因组显着或 LOD≥6和支撑间隔的暗示性QTL通常<2 mb。引人注目的定量性状区域（QTL） 饮酒的第一周与消费最后一周的饮酒不同。正在进行的单倍型分析 来自前额叶皮层的RNASEQ数据的集成已确定临时候选基因，包括 在人类全基因组饮酒或饮酒或 依赖。我们在这种续签中假设该行为QTL和基因组数据的扩展将 确定新颖的候选基因和基因网络，促进小鼠乙醇消耗行为 这样的数据将为现有的和未来的人类遗传研究和对AUD的治疗工作提供信息。 因此，我们的具体目的描述了：1）伏隔核的进一步表达遗传分析，等位基因特异性 表达分析和染色质3D构象分析，以识别和完善位置列表 与初始乙醇摄入相关的行为QTL的候选基因与渐进式消费相关； 2） 在200只小鼠的前额叶皮层和伏隔核中RNASEQ数据的基因网络分析 确定网络和可能与第一次和区分的消费紧密相关的机制 上星期; 3）验证候选基因或网络中心作为乙醇行为的功能，包括 使用无脊椎动物模型的初始或进行性乙醇消耗（与项目2和3的合作 该中心建议），啮齿动物模型（该项目和啮齿动物行为核心）。此外，这项工作 项目将通过项目4和5中描述的新型人类遗传研究告知并告知。 该项目，VCU ARC生物信息学和分析核心将为RNASEQ分析提供关键的支持 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 新颖的基因发现和网络分析工作将与VCU的所有其他组成部分进行主要相互作用 ARC并将通过了解过渡到过渡到的机制，并将其告知酒精研究领域 虐待饮酒以及未来治疗努力靶向潜在靶向的候选基因/机制。