m6A甲基化修饰调控变应性鼻炎上皮细胞中TSLP表达的机制研究

变应性鼻炎（AR）是2型免疫应答占优势的呼吸道炎性疾病，上皮源性细胞因子TSLP可促进其发生发展，但TSLP调控机制不明。m6A甲基化作为表观遗传学的重要机制，因具有动态、可逆的特性，成为当前临床转化研究的热点，但TSLP是否受m6A甲基化调控至今未有报道。申请人前期研究结果显示：AR上皮中TSLP、甲基化转移酶METTL3、结合蛋白IGF2BP1基因和蛋白水平表达均较正常对照组高，且TSLP表达水平与后两者表达均呈正相关；此外，敲低METTL3和IGF2BP1，TSLP表达可随之显著降低。据此推测：AR上皮细胞中TSLP因受到m6A甲基化修饰调控，使其表达增多，进而加重AR的2型炎症反应。本课题拟通过体外细胞培养、甲基化RNA免疫共沉淀等技术，通过人体组织、细胞及动物层面研究，进一步揭示m6A甲基化调控TSLP表达在AR中的作用机制，为探寻更理想的AR对因治疗靶点提供理论基础。

Allergic rhinitis (AR) is a respiratory inflammatory disease dominated by type 2 immune response. Epithelial derived cytokine TSLP is closely related to its pathogenesis, but the specific mechanism is unclear. As an important epigenetic mechanism, m6A methylation has become a hot spot of clinical translational research due to its dynamic and reversible characteristics. However, whether TSLP is regulated by m6A methylation has not been reported. Our previous research showed that the gene and protein expression levels of TSLP, methyltransferase METTL3 and binding protein IGF2BP1 in AR epithelium were higher than control group, and TSLP was positively correlated with the METTL3 and IGF2BP1. In addition, TSLP expression was significantly reduced by knocking down METTL3 and IGF2BP1. Therefore, we speculated that TSLP in AR epithelial cells is regulated by m6A methylation, which further aggravates the type 2 inflammatory response in AR. The aim of this study is to further reveal the mechanism of m6A methylation in regulating TSLP expression in AR. We plan to use cell culture, methylation RNA immunoprecipitation study to explore the mechanism of m6A methylation in regulating TSLP expression, and to provide a theoretical basis for exploring a more ideal therapeutic target of AR.