Association and Function of Opioid Receptor Gene Variants to Substance Dependence

阿片受体基因变异与物质依赖性的关联和功能

• 批准号: 8120382
• 负责人: Huiping Zhang
• 金额: $ 23.61万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8120382
• 关键词: Address; Affect; African American; Alcohols; American; Biological Assay; Cocaine; DNA; DNA Resequencing; Data; Epigenetic Process; European; Future; Genes; Genetic Variation; Heroin; Linkage Disequilibrium; Mediating; Modification; Morphine; Nucleic Acid Regulatory Sequences; Opioid Peptide; Opioid Receptor; Outcome; POMC gene; Peptide Receptor; Pharmaceutical Preparations; Phase; Polymerase Chain Reaction; Prevention strategy; Receptor Gene; Reporter Genes; Rewards; Substance Addiction; Techniques; Testing; Time; Variant; Western Blotting; case control; design; effective therapy; gel mobility shift assay; improved; next generation; receptor binding

Tfiis is a K99 to ROO transition application which seeks further support for studying the association of four opioid receptor genes (0PRM1, 0PRD1, 0PRK1, and 0PRL1) and four opioid peptide genes (POMC, PDYN, PENK, and PNOC) genes and substance dependence (SD) in African Americans (AAs) and European Americans (EAs). In the K99 phase, we identified several opold receptor and peptide gene variants which were significantly associated with SD. However, since only a limited number of variants were selected and tested for their association with SD, the causal variant might have been ignored. In addition, it is unknown whether the identified SD-associated variants are functional or they are just in linkage disequilibrium with the causal variant. Therefore, in the ROO phase, we need to further address these Issues. We propose to (1) analyze the association of both common and rare variants with SD by resequencing the opioid receptor and peptide genes in our cases and controls using the next-generation sequencing technique; (2) examine the function of SD-assoclated opioid receptor and peptide gene variants using several approaches including receptor binding assays, Western blotting, real-time quantitative polymerase chain reaction, allelic expression Imbalance assay, electrophoretic mobility gel shift assay, and liclferase reporter gene assay; and (3) Investigate whether DNA metylatlon levels In the regulatory regions ofthe opioid receptor and peptide genes are significantly different between SD affected cases and healthy controls. The proposed study will improve our understanding about the Influence of opioid receptor and peptide gene variants on SD and determine whether epigenetic modification ofthese genes can Increase vulnerability to SD. In addition, it will generate sufficient data for a future ROI project aimed at (1) establishing a set of opiod receptor and peptide gene markers as predictors of SD; and (2) investigating the contribution of variants In these genes to the outcome of SD treatment.

Tfiis 是一个 K99 到 ROO 的过渡应用程序，旨在为研究四项关联寻求进一步支持 阿片受体基因（0PRM1、0PRD1、0PRK1 和 0PRL1）和四种阿片肽基因（POMC、 PDYN、PENK 和 PNOC）基因与非裔美国人 (AA) 和物质依赖 (SD) 欧洲裔美国人 (EA)。 在K99阶段，我们鉴定了几种opold受体和肽基因变异体，这些变异体显着 与SD有关。然而，由于仅选择并测试了有限数量的变体 与 SD 的关联，因果变异可能被忽略。此外，尚不清楚是否 已识别的 SD 相关变异是功能性的，或者它们只是与因果关系处于连锁不平衡状态 变体。因此，在原产地规则阶段，我们需要进一步解决这些问题。我们建议（1）分析 通过对阿片受体和肽进行重新测序来确定常见和罕见变异与 SD 的关联 使用下一代测序技术对我们的病例和对照中的基因进行分析； (2)检查功能 使用多种方法（包括受体）的 SD 相关阿片受体和肽基因变体 结合测定、蛋白质印迹、实时定量聚合酶链反应、等位基因表达 不平衡测定、电泳迁移率凝胶位移测定、以及Liclferase报告基因测定；和（3） 研究阿片受体和肽基因调控区域中的 DNA 甲基化水平是否 SD 受影响病例和健康对照之间存在显着差异。 拟议的研究将增进我们对阿片受体和肽影响的理解 SD 上的基因变异并确定这些基因的表观遗传修饰是否可以增加 SD 的脆弱性。此外，它将为未来的 ROI 项目生成足够的数据，该项目旨在 (1) 建立一套阿片受体和肽基因标记作为 SD 的预测因子； (2) 调查 这些基因中的变异对 SD 治疗结果的贡献。