冠心病患者PCI术后支架内再狭窄（ISR）严重影响预后，支架植入后局部炎症所致血管平滑肌（VSMC）增殖是其主要机制，多种药物通过上调AMPK的活性而抑制VSMC增殖，降低ISR的发生风险。本项目前期研究发现AMPKα2亚单位编码基因PRKAA2遗传多态性可降低ISR发生风险的基础上，拟进一步扩大样本验证其与ISR易感性的关系，并应用微小基因和报告基因实验对PRKAA2遗传变异位点进行功能分析；同时应用CRIPSPR/CAS9、免疫共沉淀和质谱分析等技术，查明PRKAA2及其多态性是否通过影响mTOR通路和TET2介导的5-hmC生成，从而影响VSMC的增殖及其对雷帕霉素的反应性，参与ISR的发病机制。该研究旨在查明PRKAA2遗传多态性对冠心病患者PCI术后ISR易感性和雷帕霉素反应性的影响及分子机制，发现ISR新的遗传标志物，为ISR的个体化预防和防治药物的研发提供新思路。

In-stent restenosis (ISR) disease after PCI for patients suffered from coronary heart disease affects the disease prognosis. Proliferation of vascular smooth muscle cells induced by Inflammation caused by stent implantation is the main mechanism for ISR. Several medications are reported to decrease the risk of ISR by promoting the activity of AMPK and thus inhibits VSMC proliferation. In our former studies, we observed that genetic polymorphism in PRKAA2 that encoding the a2 catalytic subunit of AMPK was associated with decreased ISR risk. Based on these previous findings, the project is designed to make clear the association of PRKAA2 polymorphisms with ISR risk by enlarging sample size. And also, we aimed to perform functional analysis of PRKAA2 positive associated polymorphisms by using minigene splicing test and reporter gene assays. By using technologies such as CRIPSPR/Cas9 edition, gene intervention and overexpression, co-immunoprecipitation (CO-IP) and mass spectrometry, we also aimed to explore whether PRKAA2 functions through stimulation phosphorylation of mTOR and TET2, and thus regulating 5-hmC contents in genes involved in the maintenance of VSMC phenotypes, and consequently affecting VSMC proliferation and response to the mTOR inhibitor rapamycin. The project will help identify new genetic markers which could be used to predict the occurrence of ISR after PCI, and provide foundation for the prevention of ISR, as well as provide new idea for new drug development for the prevention of ISR.