Functional Validation of Candidate Genes Deriving from Convergent Analysis of Gene Expression Profiling in Nicotine Self-Administering Animals and Genome Wide Association Studies in Smokers

背景资料：吸烟行为遗传性的一致证据导致了旨在阐明尼古丁成瘾所涉及的特定遗传因素的遗传研究。解决这个问题的最先进的科学方法包括吸烟者的全基因组关联研究（GWAS）和尼古丁自我给药动物模型的基因表达谱研究。聚合翻译基因组学方法允许将来自动物模型的遗传发现与人类中的GWAS方法相结合，并且已被证明在生成优先候选基因列表方面非常成功。然而，候选基因与给定病理表型的因果关系的证明仍然是新兴遗传学研究领域的最大挑战。目标：在本提案中，我们希望将我们之前在接受长期尼古丁自我给药的动物中获得的大量基因表达谱的遗传学发现与来自德国尼古丁研究项目优先计划（项目P2（Dahmen），P3（Winterr），P9（Brenner），P10（Winterr）和P11（Steinlein））的人类遗传数据集相结合。通过应用聚合翻译基因组学方法，将进行数据整合，并生成整个研究项目的优先候选基因列表。从这个尼古丁成瘾的优先基因列表中，我们将通过重组腺相关病毒（rAAV）介导的基因转移到奖励系统的大脑部位（特别是进入核壳），在尼古丁自我给药和复发的动物模型中功能验证最高命中。携带针对特定基因的cDNA或针对特定基因的短发夹RNA的AAV载体的出现现在首次允许在尼古丁自我施用动物中快速双向操纵基因功能。这两个申请人分别在成瘾行为的动物模型和rAAV介导的基因转移技术方面拥有成熟的专业知识。

Background: Consistent evidence for the heritability of smoking behaviours has led to genetic studies designed to elucidate the specific genetic factors involved in nicotine addiction. The most advanced scientific approaches to address this question include genome-wide association studies (GWAS) in smokers and gene expression profiling studies in animal models of nicotine self-administration. Convergent translational genomics approaches allow integrating genetic findings from animal models with a GWAS approach in humans and have proven to be very successful in generating priority candidate gene lists. However, the demonstration of a causal relationship of a candidate gene with a given pathological phenotype remains the biggest challenge in the emerging field of genetic research. Goals: In this proposal we want to integrate our previous genetic findings from massive gene expression profiling in animals that underwent chronic nicotine self-administration with the human genetic data sets deriving from the German Priority Programme on Nicotine Research Project (Project P2 (Dahmen), P3 (Winterer), P9 (Brenner), P10 (Winterer), and P11 (Steinlein)). By applying a convergent translational genomics approach data integration will be performed and a priority candidate gene list for the entire Research Project will be generated. From this priority gene list for nicotine addiction we will functionally validate the top hits in an animal model of nicotine self-administration and relapse by means of recombinant adeno-associated virus (rAAV)-mediated gene transfer into brain sites of the reward system (especially into the nucleus accumbens shell). The advent of AAV vectors carrying cDNA for, or short hairpin RNA against, specific genes allows now for the first time the rapid bidirectional manipulation of gene function in nicotine self-administering animals. Both applicants have proven expertise in animal models of addictive behaviour and rAAV-mediated gene transfer technology, respectively.