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

项目摘要摘要 流行病学研究表明，基因变异对止痛药的作用显著(20-60%)。 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(GWASs)才刚刚开始识别导致这种风险的特定基因。一个主要的障碍是 阿片类药物使用障碍的研究是表型的复杂性和缺乏对环境变量的控制。 我们提出了一种互补的方法，利用一个多学科、高度协作的财团 将下一代测序与最先进的行为筛查相结合，在一种独特的、遗传的 多样化的非人类动物模型。这项提议的主要目标是识别出 与强迫性羟考酮使用的易感性相关，对羟考酮止痛效果的耐受性 羟考酮，戒断诱导的痛觉过敏的发展，以及对FDA批准的药物的敏感性 在N/NIH异种系大鼠上进行大鼠肾间质干细胞移植。我们将使用最相关的动物模型 羟考酮使用障碍(即静脉注射羟考酮自我给药升级)和高度标准化 强迫性羟考酮自我给药与纵向疼痛评估相结合 阈值。这个项目可能会对该领域产生持续而强大的影响，因为它将(1) 从控制使用羟考酮到强制使用羟考酮的转变及其与痛觉过敏的共病 在雄性和雌性杂交大鼠中，(2)确定与强迫性羟考酮使用相关的基因，临床前 当前药物(如丁丙诺啡)的疗效以及慢性前列腺癌的止痛/过敏性作用 羟考酮的使用，以及(3)通过创建包含大脑和血液的储存库来促进后续研究 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、生化和 行为/遗传特征动物的药理学研究。

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.