Genetics of oxycodone intake in a hybrid rat diversity panel.

羟考酮等处方阿片类药物的稳步增长导致了美国广泛的滥用和死亡。 药物药代动力学在确定滥用潜力中的重要性，我们设计了一种口服药物 模拟大多数人类吸毒者/吸毒者药物摄入模式的可操作性大鼠自我给药(SA)程序 羟考酮，开始使用口服片剂。尽管基因变异在易感性上起着重要作用 阿片成瘾，有关易患阿片成瘾的特定基因和序列变异的数据非常有限 对阿片成瘾，以及在什么条件下。 给定 我们建议使用一种创新的杂交老鼠多样性小组 (HRDP)，由91个不同的大鼠基因组组成，以识别影响操纵性羟考酮的遗传变异 大鼠的摄入量。HRDP的独特之处在于：1)它含有与人类相似的高度遗传多样性 种群；2)提供了一种控制羟考酮暴露和系统研究基因与环境的方法 以及基因与药物的相互作用；以及3)整合多组学“专有”数据：从遗传学到表观基因组学，再到 大脑与治疗的联系。我们有三个目标：目标1：分析全基因组测序数据 以定义几乎所有构成可遗传变异基础的序列变异。将进行新的组装 使用连锁-读取选定高影响菌株的数据。将生成HI-C数据(Dovetail基因组学)以 进一步提高这些组件的质量。我们还将为关键的大脑区域生成RNA-SEQ数据 获得羟考酮摄入的机械性见解。目的2：使用HRDP(两性)，我们将进行口腔表型 羟考酮SA具有独特的行为模式。还将测试大鼠对疼痛的敏感性，社交行为， 以及类似焦虑的特征--都是羟考酮停药的迹象。关键的是，我们估计了遗传力(H2) 羟考酮摄入量在0.3-0.4之间。当使用n=6/性别时，有效H_2为~0.8-足以满足高 精确测绘。目标3：我们将使用系统遗传学方法来映射和整合行为表型 序列和转录组数据。正向(QTL)和反向(Phewas)两种遗传方法都将被使用。 我们使用新的线性混合模型通过GeneNetwork2来定位和测试带有关键辅助因子的候选基因 站台。最后，我们通过比较来评估候选基因和生物标志物的翻译相关性 关于人类成瘾的GWA群和纵向报告。技术和概念上的进步 这种应用是：新的基因组方法与高度多样化的老鼠种群相结合，使我们能够迅速 定义调节阿片成瘾关键阶段的新基因变种。很可能是变种的一个子集 我们定义的分子网络将提供预测框架连接序列的关键组件 与人类阿片成瘾的差异和可能的治疗方法。这个项目使用了新的系统遗传学 方法，开源基因组数据和软件，以及一种新型的杂交啮齿动物地图绘制小组 准确定义DNA变异和自愿羟考酮摄入量之间的因果联系。

The steady rise in prescription opioids such as oxycodone has led to widespread abuse and deaths in the US. importance of drug pharmacokinetics in determining abuse potential, we have designed an oral operant rat self-administration (SA) procedure to model the pattern of drug intake of most human users/abusers of oxycodone, who initiate using oral tablets. Although genetic variants play important roles in susceptibility to opioid addiction, very limited data are available regarding specific genes and sequence variants that predispose to opioid addiction, and under what conditions. Given the We propose to use an innovative hybrid rat diversity panel (HRDP), which consists of 91 diverse rat genomes, to identify genetic variants influencing operant oxycodone intake in rats. The HRDP is unique in that it: 1) contains a high level of genetic diversity similar to that of human populations; 2) provides a way to control oxycodone exposure and to systematically study gene-by-environment and gene-by-drug interactions; and 3) integrates multi-omics "addictome" data: from genetics to epigenomics to brain connectomes to treatments. We have three aims: Aim 1: We will analyze whole genome sequencing data to define virtually all sequence variants that underlie heritable variations. De novo assemblies will be conducted using linked-reads data for selected high impact strains. Hi-C data (Dovetail Genomics) will be generated to further improve the quality of these assemblies. We will also generate RNA-seq data for key brain regions to obtain mechanistic insights into oxycodone intake. Aim 2: Using the HRDP (both sexes), we will phenotype oral oxycodone SA with a unique behavioral model. Rats will also be tested for sensitivity to pain, social behaviors, and anxiety-like traits - all signs of oxycodone withdrawal. Critically, we estimated the heritability (h2) of oxycodone intake in the range of 0.3 – 0.4. When using n=6/sex, the effective h2 is ~0.8 —sufficient for high precision mapping. Aim 3: We will use systems genetics methods to map and integrate behavioral phenotypes with sequence and transcriptome data. Both forward (QTL) and reverse (PheWAS) genetic methods will be used. We use new linear mixed models to map and test candidate genes with key cofactors using the GeneNetwork2 platform. Finally, we evaluate the translational relevance of candidate genes and biomarkers by comparison to GWAS cohorts and longitudinal reports of addiction in humans. Technical and conceptual advances that underlie this application are: new genomic methods combined with highly diverse rat populations allow us to quickly define novel gene variants that modulate key phases of opiate addiction. It is highly likely that a subset of variants and molecular networks we define will provide key components of a predictive framework linking sequence differences to human opioid addiction and potential treatments. This project uses new systems genetics approaches, open source genomic data and software, and a new type of hybrid rodent mapping panel to precisely define causal linkages between DNA variation and voluntary oxycodone intake.