代谢酶基因组eQTL对氯吡格雷活性代谢物生成的影响及其功能评价

氯吡格雷活性代谢物（AM）生成的个体差异是氯吡格雷抗血小板疗效个体差异的关键来源。我们前期研究及发表数据显示，已知的基因变异不足以解释氯吡格雷AM生成的个体差异，这与以往的研究纳入的功能性基因变异数量不完全有关，甚至有一半以上基因表达数量性状位点（eQTL）没有被纳入。我们推测发现原因性eQTL可提高对氯吡格雷AM生成个体差异及临床不良终点事件的预测。本研究拟在以往研究的基础上，针对代谢酶基因组eQTL富集区域，用目标区域捕获测序和贯穿分析等方法发现影响氯吡格雷AM生成的eQTL；然后在冠心病患者中评价因果关系最强的热点区域eQTL对氯吡格雷AM药动学和疗效中间指标的影响，及对临床终点事件的预测作用；进一步用等位基因不平衡表达分析和荧光素酶报告基因技术等评价eQTL对基因表达的调控作用。本研究将有助于深入了解代谢酶基因变异功能，并有助于危险分层，优化抗血小板给药方案，减少临床终点事件。

The individual difference in the formation of clopidogrel active metabolites (AM) is the major source for the individual difference in efficacy of clopidogrel antiplatelet treatment. Published data and our primary data indicated that current known genetic variations were far from sufficient to explain the individual difference in the formation of clopidogrel AM. One of the reasons is that many functional variations were not included, even more than half of expression quantitative trait loci (eQTL) were missed in previous studies. We hypothesize that the discovery of eQTL will improve the predictivity of the individual difference in the formation of clopidogrel AM and occurrence of adverse clinical endpoints. In the proposal, based on previous results, targeted next-generation sequencing and the Network Edge Orienting (NEO) method will be applied to discover causal eQTLs for the formation of clopidogrel AM. The effect of eQTLs in hotspot with the most strong causal relationship with the formation of clopidogrel AM on the pharmacokinetics and antiplatelet efficacy of clopidogrel in patients with coronary artery disease will be evaluated. A risk predictive model with eQTL genotypes will thereafter be developed. Several techniques, including allelic expression imbalance analysis and luciferase Reporter Gene Assays, will further be applied to illustrate the regulation mechanism of eQTL on gene expression. The results of this study will give us insight into the function of genetic variation in metabolic enzyme genes and will help clinic optimize the antiplatelet therapy strategy of clopidogrel in patients with coronary artery disease after percutaneous coronary intervention.