内淋巴囊上皮细胞源外泌体通过转运miR-222/MUC1调控免疫应答在AIED 发病中的作用及机制研究

内淋巴囊（ES）介导免疫紊乱是自身免疫性内耳病（AIED）发病的潜在机理。ES主要由上皮细胞和散在其上或周围的免疫活性细胞组成。提示：上皮细胞和免疫细胞之间存在互作，然而机制尚未阐明。本课题组前期ES表达谱分析发现MUC1是ES上皮细胞中关键基因，而AIED模型鼠miRNAs表达谱分析鉴定miR-222-3p靶向调控MUC1。文献证明MUC1可活化促炎免疫细胞，而外泌体是细胞交流的重要介质。据此，推测ES上皮细胞通过外泌体将MUC1带入免疫细胞，调节炎症因子释放和外周免疫细胞归巢，而外泌体miR-222-3p减少是MUC1增多的潜在原因。为此，本课题首先对ES上皮细胞外泌体进行测序，确定外泌体MUC1和miR-222-3p表达；其次，将外泌体和免疫细胞共培养，确定外泌体对免疫细胞的影响；再次，过表达miR-222-3p和MUC1抑制剂外泌体注射AIED模型鼠，确定其对AIED的治疗作用。

Endolymphatic sac (ES) mediated immune disorder is an underlying pathogenesis of the development of autoimmune inner ear disease (AIED). ES is composed of epithelial cells and immunocompetent cells within or around it, suggesting there may be an interaction between ES epithelial cells and immunocompetent cells, but the interaction mechanisms remain unclear. Our previous study investigated the mRNA expression profile of ES showed MUC1 may be crucial genes in ES epithelial cells, the miRNA expression profile indicated miR-222-3p could regulate MUC1. The literatures demonstrated MUC1 could activate the proinflammatory immune cells, while exosomes are important mediators for cell communication. Accordingly, we speculate ES epithelial cells may carry the MUC1 to immune cells by exosomes and then mediate the release of proinflammatory cytokines and the homing of peripheral immune cells, while the lower exosomal miR-222-3p may be the cause for the increase in MUC1. To prove our hypothesis, we aim to first sequence the exosomes of ES epithelial cells to confirm the expression of MUC1 and miR-222-3p; then, co-culture the exosomes with the immune cells to validate the influence; injection of exosomal miR-222-3p and anti-MUC1 to explore their therapeutic effects for AIED.