Genetics of oxycodone intake in a hybrid rat diversity panel.

处方阿片类药物如羟考酮的稳步上升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的重要性，我们设计了一种口服 操作性大鼠自我给药（SA）程序，以模拟大多数人类使用者/滥用者的药物摄入模式 开始使用口服片剂的人。虽然遗传变异在易感性方面发挥着重要作用， 阿片类药物成瘾，非常有限的数据是关于特定的基因和序列变异，易患 阿片类药物成瘾，以及在什么情况下。 鉴于 我们建议使用创新的混合鼠多样性小组 （HRDP），由91个不同的大鼠基因组组成，以确定影响操作性羟考酮的遗传变异 大鼠摄入量。HRDP的独特之处在于：1）它含有与人类相似的高水平遗传多样性 人群; 2）提供了一种控制羟考酮暴露和系统研究基因环境的方法 以及基因与药物的相互作用;以及3）整合多组学“成瘾组”数据：从遗传学到表观基因组学， 大脑连接体与治疗的联系我们有三个目标：目标1：我们将分析全基因组测序数据 来定义几乎所有的序列变异，这些变异是遗传变异的基础。将进行重新组装 使用选定的高冲击应变的连接读数数据。将生成Hi-C数据（Dovetail Genomics）， 进一步提高这些组件的质量。我们还将生成关键大脑区域的RNA-seq数据， 获得对羟考酮摄入的机械见解。目的2：使用HRDP（两性），我们将表型口腔 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，社会行为， 和焦虑症状都是羟考酮戒断症状关键的是，我们估计了遗传力（h2）， 羟考酮摄入量在0.3 - 0.4的范围内。当使用n=6/性别时，有效h2约为0.8 -足以用于高 精确制图目标3：我们将使用系统遗传学方法来绘制和整合行为表型 序列和转录组数据。将使用正向（QTL）和反向（PheWAS）遗传方法。 我们使用新的线性混合模型来映射和测试候选基因与关键辅因子使用GeneNetwork 2 平台最后，我们通过比较候选基因和生物标志物的翻译相关性， GWAS队列和人类成瘾的纵向报告。技术和概念上的进步， 该应用是：新的基因组方法与高度多样化的大鼠种群相结合，使我们能够快速 定义新的基因变异，调节阿片类药物成瘾的关键阶段。很有可能一部分变异体 我们定义的分子网络将提供预测框架连接序列的关键组成部分 人类阿片类药物成瘾和潜在治疗的差异。这个项目使用新的系统遗传学 方法，开源基因组数据和软件，以及一种新型的混合啮齿动物绘图面板， 精确定义DNA变异和自愿羟考酮摄入之间的因果关系。

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.