Use of Next-Gen Sequencing to Identify Genetic Variants that Influence compulsiveOxycodone Intake in Outbred Rats

项目摘要 流行病学研究表明，遗传变异性显着贡献（20-60%）的止痛药， 对阿片类药物的反应和对阿片类药物使用障碍的脆弱性。然而，全基因组关联研究 人类的GWASs（GWASs）只是开始识别赋予这种风险的特定基因。一个主要障碍是 阿片类药物使用障碍的研究是复杂的表型和缺乏控制的环境变量。 我们提出了一种互补的方法，利用多学科，高度合作的财团， 将下一代测序与最先进的行为筛查相结合， 多样的非人类动物模型。这项提案的主要目标是确定基因变异， 与更容易强迫使用羟考酮、耐受羟考酮的镇痛作用有关， 羟考酮、戒断诱导的痛觉过敏的发生以及对FDA批准药物的敏感性， 在N/NIH异质储备大鼠中进行GWAS。我们将使用最相关的动物模型， 羟考酮使用障碍（即，静脉内羟考酮自我给药的升级）和高度标准化 强制性羟考酮自我给药的测量与疼痛的纵向评估相结合 阈值。该项目可能会对该领域产生持续而强大的影响，因为它将（1） 描述羟考酮使用从控制到强制的转变及其与痛觉过敏的共病性 在雄性和雌性远系繁殖大鼠中，（2）鉴定与强迫性羟考酮使用相关的基因， 当前药物的功效（例如，丁丙诺啡），以及慢性丁丙诺啡的镇痛/痛觉过敏作用 羟考酮的使用，和（3）通过创建一个包含大脑和血液的储存库， 各种组织保存方案，通过允许 诱导多能干细胞的产生和神经解剖学、分子学、生物化学和 对行为/遗传特征动物进行药理学研究。

Project SummaryAbstract Epidemiological studies suggest that genetic variability significantly contributes (20-60%) to the analgesic response to opioids and vulnerability to opioid use disorder. However, genome-wide association studies (GWASs) in humans have only begun to identify specific genes that confer this risk. One major impediment to studies of opioid use disorder is the complexity of the phenotype and lack of control of environmental variables. We propose a complementary approach that leverages a multidisciplinary, highly collaborative consortium that combines next-generation sequencing with state-of-the-art behavioral screening in a unique, genetically diverse, nonhuman animal model. The primary goal of this proposal is to identify gene variants that are associated with greater vulnerability to compulsive oxycodone use, tolerance to the analgesic effects of oxycodone, development of withdrawal-induced hyperalgesia, and sensitivity to FDA-approved medications by performing a GWAS in N/NIH heterogeneous stock rats. We will use the most relevant animal model of oxycodone use disorder (i.e., escalation of intravenous oxycodone self-administration) and highly standardized measures of compulsive oxycodone self-administration combined with longitudinal assessments of pain thresholds. This project is likely to have a sustained and powerful impact on the field because it will (1) characterize the transition from controlled to compulsive oxycodone use and its comorbidity with hyperalgesia in male and female outbred rats, (2) identify genes associated with compulsive oxycodone use, the preclinical efficacy of current medication (e.g., buprenorphine), and the analgesic/hyperalgesic effects of chronic oxycodone use, and (3) facilitate follow-up studies by creating a repository that contains brain and blood with a variety of tissue preservation protocols that will facilitate follow-up and replicative studies by allowing the generation of induced pluripotent stem cells and neuroanatomical, molecular, biochemical, and pharmacological studies on behaviorally/genetically characterized animals.