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

基本信息

批准号：
7496083
负责人：
Huiping Zhang
金额：
$ 8.85万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-15 至 2009-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7496083
关键词：
Adverse effects Affect Affinity African American Alcohol dependence Alcohols Alleles American Appendix Binding Binding Sites Biological Assay Buprenorphine Clinical Clinical Trials Code Complex DNA Sequence Diagnosis Disease Early Diagnosis Early Intervention Economics Effectiveness Electrophoretic Mobility Shift Assay Environmental Risk Factor Ethnic group European Family Functional RNA Future Gene Expression Genes Genetic Genetic Models Genetic Variation Genotype Goals Haplotypes Human Individual Lead Luciferases Mediating Mentors Messenger RNA Minor Naloxone Opiate Addiction Opioid Opioid Receptor Outcome Patients Personal Satisfaction Pharmaceutical Preparations Pharmacotherapy Phase Phase I Clinical Trials Phenotype Polymerase Chain Reaction Population Proteins Range Receptor Gene Recruitment Activity Reporter Genes Research Personnel Research Training Rewards Risk Safety Single Nucleotide Polymorphism Structure Substance Addiction Testing Time Training Treatment outcome Variant Western Blotting Withdrawal Work base case control dosage experience genetic association improved interest neuropsychiatry non-opioid analgesic programs psychosocial racial difference receptor receptor binding response transcription factor

项目摘要

DESCRIPTION (provided by applicant): This K99/ROO application seeks support for additional research training which will enable the applicant to become an independent investigator in neuropsychiatric genetics. The training goal will be achieved through a project aimed to identify substance (drug and/or alcohol) dependence (SD)-associated variants in three opioid receptor genes (OPRM1, OPRD1 and OPRK1) and interpret the mechanism of the association. In the mentored phase (year 2007-2009), I will use population- and family-based approaches to study the possible association between opioid receptor gene (OPR) variants and SD in both European Americans (EAs) and African Americans (AAs). Tightly-spaced single nucleotide polymorphisms (SNPs) spanning OPRs will be genotyped and the association between OPR variants and SD will be analyzed by appropriate statistical programs including structured association analysis. Moreover, gene regions of interest will be screened for new variants, which will be further analyzed for their association with SD. In the independent phase (year 2009-2012), I will focus on functional study of those OPR variants which are found to be associated with SD. For coding region variants, functional study will be conducted by receptor binding assay (to see if OPR variants alter receptor affinity) and Western Blotting (to see if OPR variants modulate receptor protein levels). For non-coding region variants, functional study will be performed by four different approaches: (1) real-time quantitative PCR, which examines whether OPR variants result in increased or decreased gene expression (or mRNA) levels; (2) allelic expression imbalance (AEI) assay, which is an alternative approach to examine whether the two alleles of a variant lead to different expression (or mRNA) levels; (3) Electrophoretic mobility shift assay (EMSA), which examines whether OPR variants are located in transcription factor (TF) binding sites and if they change the binding of a TF to its DNA sequence; (4) luciferase reporter gene assay, which examines whether OPR variants regulate gene expression. These four approaches are complementary. The proposed study will improve our understanding about the influence of OPR variants on SD in EAs and AAs. This work will lay the groundwork for a future R01 project aimed to establish a genetic model for prediction of SD with a set of OPR markers, and to investigate the contribution of OPR variants to the outcome of drug or alcohol dependence treatment or to the racial difference in outcomes of treatment. The applicant is mentored by Dr. Joel Gelernter (primary) and Dr. Jeffrey Gruen (co-mentor), who are both excellent supervisors with substantial experience in genetic studies of complex disorders and who have both mentored numerous previous trainees.

描述（由申请人提供）：此K99/ROO申请寻求支持其他研究培训，这将使申请人成为神经精神遗传学的独立研究者。培训目标将通过旨在识别三个阿片类药物受体基因（OPRM1，OPRD1和OPRK1）的物质（药物和/或酒精）依赖性（SD）依赖性（SD）相关的项目实现，并解释了该关联的机制。在指导阶段（2007- 2009年），我将使用基于人群和家庭的方法研究欧美人（EAS）和非裔美国人（AAS）中阿片受体基因（OPR）变体（OPR）变体之间可能的关联。将对跨越OPRS的紧密间隔单核苷酸多态性（SNP）进行基因分型，并且OPR变体和SD之间的关联将通过包括结构化关联分析在内的适当统计程序进行分析。此外，将筛选新变体的感兴趣基因区域，这将进一步分析其与SD的关联。在独立阶段（2009 - 2012年）中，我将重点研究与SD相关的那些OPR变体的功能研究。对于编码区域的变异，功能研究将通过受体结合测定（查看OPR变体改变受体亲和力）和蛋白质印迹（查看OPR变体是否调节受体蛋白水平）来进行功能研究。对于非编码区域的变体，功能研究将通过四种不同的方法进行：（1）实时定量PCR，它检查了OPR变体是导致基因表达（或mRNA）水平升高还是下降；（2）等位基因表达不平衡（AEI）测定，这是检查变体的两个等位基因是否导致不同表达（或mRNA）水平的另一种方法；（3）电泳迁移率转移测定法（EMSA），它检查了OPR变体是否位于转录因子（TF）结合位点以及它们是否改变TF与其DNA序列的结合；（4）荧光素酶报告基因测定法，该测定法检查了OPR变体是否调节基因表达。这四种方法是互补的。拟议的研究将提高我们对OPR变体对EAS和AAS中SD的影响的理解。这项工作将为未来的R01项目奠定基础，旨在建立一个用一组OPR标记来预测SD的遗传模型，并研究OPR变体对药物或酒精依赖治疗结果的贡献或治疗结果对种族差异的贡献。申请人由Joel Gelernter博士（小学）和Jeffrey Gruen博士（Co-Ins-Encortor）指导，他们都是出色的主管，在复杂疾病的遗传研究方面拥有丰富的经验，并且都曾指导过许多以前的学员。