DANCR201/miR-216a/端粒酶激活的信号轴调控动脉粥样硬化血管衰老和斑块稳定性的分子与临床研究

血管老化是动脉粥样硬化的重要特征。前期工作通过全转录组测序发现lncRNA DANCR201在冠脉混合斑块组患者低表达，且在老年患者外周血中表达水平显著降低；细胞定位在斑块部位内皮细胞，并发现DANCR201竞争结合miR-216a，调控下游靶点端粒酶活性。基于此，提出科学假设：DANCR201/miR-216a/端粒酶信号轴调控血管老化和斑块进展。本项目拟解决问题1: 通过基因过表达、siRNA阻断技术以及内皮细胞特异敲除DANCR201-/-、miR-216a-/-小鼠模型，明确DANCR201通过ceRNA方式募集miR-216a，调控端粒酶活性，进而影响动脉粥样硬化血管衰老和斑块稳定的分子机制；2：检测冠脉钙化前瞻性队列外周血DANCR201和miR-216a表达与血管内皮功能及斑块特征的关系。这将为阐明lncRNAs调控血管内皮炎性衰老和动脉粥样硬化斑块稳定性的机制提供新靶点。

Endothelial senescence related vessel dysfunction is important risk factor for the development of atherosclerosis and cardiovascular diseases. Recently, long noncoding RNA (lncRNAs) has aroused growing interests in life sciences, but the role of which in atherosclerotic vessel dysfunction remains largely unknown. ..With the use of whole-blood transcriptome sequencing technique, our preliminary data identified a new lncRNA, DANCR201, which showed a significantly lower expression in patients with mixed plaques, compared with those patents with calcification plaques and healthy subjects. Stable expression of DANCR201 by lentivirus in human umbilical vein endothelial cell (HUVECs) significantly improved the cell viability such as cell proliferation, migration and inhibited monocytes adhesion to endothelial cell monolayers. In addition, analysis of miRNA-lncRNA interactions indicated that DANCR201 contains a candidate binding site of miR-216a between 764 and 770 bps. Furthermore, we also found that miR-216a triggered the endothelial senescence and promoted the endothelial inflammation by activating the NF-κB signaling pathway. Based on these interesting preliminary findings, we proposed the hypothesis that DANCR201 may regulate atherosclerotic vascular senescence and plaque instability through NF-κB/telomerase pathway by sponging miR-216a...In the present project, we aimed to (1) investigate whether DANCR201 can regulate the activation of telomerase activity by sponging miR-216a and modulate the endothelial senescence related dysfunctions in vitro and vivo, via the cellular model of lentivirus overexpression, or siRNA technique, as well as the ApoE-/- atherosclerotic model of mice with endothelial cell-specific knockdown of DANCR201-/- and miR-216a-/-；(2) investigate the relationship between circulating DANCR201, miR-216a levels and progression of coronary artery plaques and the risk of future cardiovascular diseases in a perspective cohort of coronary heart disease patients. This study will provide new targets for the diagnosis and treatment of aging-related atherosclerotic diseases.