The Genetic Basis of Opioid Dependence Vulnerablility in a Rodent Model

项目总结 阿片类药物滥用是一个严重的全球性问题，影响到所有社会的健康、社会和经济福利。 阿片类药物使用障碍(OUD)是一种以强迫使用阿片类药物为特征的医学状况，尽管存在不良反应 持续使用和阿片类药物停用后出现戒断综合症的后果。OUD 涉及对阿片类药物的上瘾和依赖。阿片类药物包括处方药，如 吗啡、可待因、羟考酮和氢可酮以及海洛因等非法药物。虽然动物模型 提供严格、方便的手段，精确控制环境背景和药物暴露，并评估 行为、基因组、生化和认知变化，有效利用这些模型作为有效的替代 因为人类的毒瘾仍然具有挑战性。调节人的行为差异的基因变体的身份 实验室动物在很大程度上仍不为人所知，极大地限制了对生理差异和 了解环境对药物滥用的影响，并阻碍将遗传研究结果转化为 人类。这一局限性突出了对健壮动物的完整基因组特征的迫切需要 阿片类药物滥用的模式。这个项目的目标是开发一种研究阿片类药物滥用的杂交动物模型， 与全面的基因组特征相结合，以改进对潜在表型和 与向阿片类药物滥用过渡相关的基因组变化。该项目将吸引一名国际 成瘾科学和行为研究、神经生物学、啮齿动物等领域的多学科专家团队 遗传学、计算基因组学、生物信息学以及高通量计算和数据建模 实现三个具体目标。在目标1中，我们将开发一种经过验证的海洛因自我繁殖动物模型。 具有扩展访问权限的给药，产生与增加的药物一致的行为表型 在戒毒期间，吸食毒品、吸食毒品的动机加快以及寻求毒品的人数增加。 两个不同地理位置的动物(一个在美国，一个在欧洲)将被分成不同的层次 “易受攻击的”“耐药”谱，目标是对所有个体进行基因筛查。在AIM 2，我们将在1,000人中使用测序基因分型(GBS)进行全基因组关联研究(Gwas 从这两个地点采集了动物样本。对最重要的遗传变异的验证将在#年进行 一个由100只动物组成的独立验证队列。在目标3中，我们将评估基因对基因组的影响 通过高通量RNA测序在AIM 2中发现变异体，以评估相关基因的表达 功能基因和发现表达的数量性状基因座(EQTL)。染色体构象捕捉(3C) 将用于将eQTL与其目标基因进行物理链接，从而将因果关系分配给变体及其受调控的基因。

PROJECT SUMMARY Opioid abuse is a serious global problem that affects the health, social and economic welfare of all societies. Opioid use disorder (OUD) is a medical condition characterized by the compulsive use of opioids despite adverse consequences from continued use and the development of a withdrawal syndrome when opioid use ends. OUD involves both addiction to and dependence upon opioids. Opioids include prescription pharmaceuticals such as morphine, codeine, oxycodone and hydrocodone as well as illicit drugs such as heroin. Although animal models provide a rigorous, convenient means to precisely control environmental context and drug exposure, and assess behavioral, genomic, biochemical and cognitive changes, effective utilization of such models as an efficient proxy for human addiction remains challenging. The identities of gene variants that mediate behavioral differences in laboratory animals remain largely unknown, greatly limiting interpretation of physiologic differences and understanding of the environmental effects on drug abuse, and hindering translation of genetic findings to humans. This limitation highlights the urgent need for an integrated genomic characterization of a robust animal model of opioid abuse. The goal of this project is to exploit an outbred animal model of opioid abuse research, coupled with comprehensive genomic characterization, to improve detection of the underlying phenotypic and genomic changes associated with transition to opioid abuse. The project will engage an international multidisciplinary team of experts in the areas of addiction science and behavioral studies, neurobiology, rodent genetics, computational genomics, bioinformatics, and high throughput computing and data modeling to accomplish three Specific Aims. In Aim 1, we will exploit a validated outbred animal model of heroin self- administration with extended access that produces behavioral phenotypes consistent with increased drug consumption, accelerated motivation for drug intake and elevated drug-seeking in periods of drug absence. Animals at two distinct geographical sites (one in the USA and one in Europe) will be stratified across a ‘vulnerable’  ‘resistant’ spectrum with the objective of performing genetic screening on all individuals. In Aim 2, we will carry out a genome-wide association study (GWAS) using genotyping by sequencing (GBS) on 1,000 animals sampled from the two locations. Validation of the most significant genetic variants will be performed in an independent validation cohort of 100 animals. In Aim 3, we will assess the genomic impact of the genetic variants uncovered in Aim 2 via high throughput RNA sequencing to assess gene expression of relevant functional genes and uncover expression quantitative trait loci (eQTLs). Chromosome conformation capture (3C) will be used to physically link an eQTL with its target gene assigning causality to a variant and its regulated gene.