m6A circZBTB44通过抑制DNA甲基化上调SOCS3水平影响钙化性主动脉瓣膜疾病进程的机制研究

钙化性主动脉瓣疾病（CAVD）是老年人常见的心脏瓣膜病变，其发病机制尚不清楚，临床上缺乏防治药物。前期研究显示白介素37通过抑制炎症和成骨反应延缓CAVD进展。通过对比分析CAVD和非CAVD患者主动脉瓣组织测序结果，发现CAVD患者瓣膜组织中m6A甲基转移酶（METTL3）表达上升，而circZBTB44和SOCS3的表达下降。然而circZBTB44是否能调控CAVD进程以及调控具体机制尚未清楚。本项目拟通过体内外实验，运用RT-PCR、免疫印迹、免疫染色、免疫共沉淀、MeRIP等方法，从人体标本、动物、细胞、基因、分子等层面上验证白介素37通过METTL3靶向调节circZBTB44的m6A修饰，circZBTB44通过抑制DNA甲基化上调SOCS3水平影响CAVD进程的科学假想。该项目将为理解CAVD发病机制和circRNA在CAVD中作用提供理论基础，并为防治该病提供新思路。

Calcified aortic valve disease (CAVD) is a most common valvular heart disease in the elderly.The pathogenesis of CAVD remains unclear, and the preventive and therapeutic drugs are still lacking. Previous studies have shown that interleukin-37 alleviated the progression of CAVD by inhibiting inflammatory and osteogenic responses. By analyzing the sequencing data of aortic valve tissue in CAVD and non-CAVD patients, we found that the expression of Methyltransferase-like protein 3 (METTL3) increased in valve tissue of CAVD patients, while the expression of circZBTB44 and SOCS3 decreased. However, it is unclear whether and how circZBTB44 plays a role in the pathogenesis of CAVD.This project intends to verify the hypotheses that interleukin-37 regulates the m6A modification of circZBTB44 through suppressing METTL3 expression and circZBTB44 up-regulates SOCS3 expression to affect the progression of calcified aortic valve disease through inhibition DNA methylation by using RT-PCR, immunoblotting, immunostaining, co-immunoprecipitation, MeRIP and other methods to perform in vivo and in vitro experiments in human samples, animals, cells, genes, moleculars. The expected results of this project will provide new insights in understanding the pathogenesis of CAVD and the role of circRNA in CAVD, as well as provide new clues for the prevention and treatment of this disease.