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ROO过渡应用的K99 阿片受体基因（0PRM1、0PRD1、0PRK1和0PRL1）和四个阿片类肽基因（POMC，POMC， 非洲裔美国人（AAS）和 欧美人（EA）。 在K99阶段，我们鉴定了几种Opold受体和肽基因变异，它们显着 与SD相关。但是，由于仅选择了有限数量的变体并测试了其 与SD的关联，可能忽略了因果变体。另外，未知是否 已识别的SD相关变体具有功能性，或者只是与因果关系不平衡 变体。因此，在ROO阶段，我们需要进一步解决这些问题。我们建议（1）分析 通过重塑阿片受体和肽，共同变体和稀有变体与SD的关联 在我们的病例和使用下一代测序技术的基因和对照中； （2）检查 使用包括受体在内的几种方法 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，电泳迁移率凝胶移位测定和liclferase报告基因基因测定； （3） 研究阿片受体和肽基因调节区域中的DNA元素水平是否存在 SD受影响的病例和健康对照之间有显着差异。 拟议的研究将提高我们对阿片类药物受体和肽影响的理解 SD上的基因变体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，它将为针对（1）的未来ROI项目生成足够的数据 建立一组OPIOD受体和肽基因标记物作为SD的预测因子； （2）调查 这些基因中变体对SD治疗结果的贡献。