不同细胞来源的CHI3L1在嗜酸粒细胞性CRSwNP发病机制中的作用

The pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) is not entirely clear now. Our previous work suggested that mucosal epithelium- and macrophage-derived chitinase 3-like 1 (CHI3L1) may play an important role in eosinophilic CRSwNP. Accordingly, we intend to ①establish eosinophilic CRSwNP mice model with CHI3L1 knockout mice, change CHI3L1 expression in the murine rhinosinal mucosal epithelium and alternatively activated macrophage by using in vivo intranasal CHI3L1 gene transfer technology and macrophage adoptive transfer technology, respectively; observe mucosal inflammatory reaction in eosinophilic CRSwNP by molecular biological, immunological and histopathological techniques; screen the down-stream targets of CHI3L1 in the pathogensis of eosinophilic CRSwNP by gene array technology; ②according to the results of the in vivo study, discover the precise mechanism of CHI3L1 in regulation of rhinosinal mucosa inflammation and the pathogenesis of eosinophilic CRSwNP by further using in vitro cell culture; ③ then confirm the results of in vivo and in vitro study in rhinosinal tissue from eosinophilic CRSwNP patients by using RT-PCR and immunohistology. This project seeks to clarify the precise role of CHI3L1 in mucosal inflammation of eosinophilic CRS, in-depth understanding of CHI3L1 function, and might be helpful for discovering novel therapeutic targets in this disease.

伴鼻息肉的慢性鼻－鼻窦炎（CRSwNP）发病机制目前还不完全清楚。我们前期的研究提示鼻黏膜上皮和巨噬细胞来源的壳多糖酶3样蛋白1（CHI3L1）可能在嗜酸粒细胞性CRSwNP中起到重要作用。据此，我们拟① 利用CHI3L1基因敲出小鼠建立嗜酸粒细胞性CRSwNP模型，采用质粒转染和过继转移，改变CHI3L1在鼻黏膜上皮和巨噬细胞中的表达；观察CHI3L1对嗜酸粒细胞性CRSwNP黏膜炎症的影响；采用基因芯片技术筛选CHI3L1下游分子；② 采用细胞培养和共培养技术，干扰CHI3L1下游分子，在体外研究CHI3L1对鼻-鼻窦黏膜炎症反应调控的精确分子机制；③ 继而在人体CRS组织中验证，确认CHI3L1在嗜酸粒细胞性CRSwNP发病机制的作用。本课题力图揭示CHI3L1在嗜酸粒细胞性CRSwNP黏膜炎症中的具体机制，深入认识CHI3L1的功能，为发现新的治疗CRSwNP药物靶点打下基础。

伴鼻息肉的慢性鼻－鼻窦炎（CRSwNP）发病机制目前还不完全清楚。本项目研究鼻黏膜上皮和巨噬细胞来源的壳多糖酶3样蛋白1（CHI3L1）在嗜酸粒细胞性CRSwNP中的作用。我们成功建立小鼠建立嗜酸粒细胞性CRSwNP模型，采用siRNA技术敲出CHI3L1基因，慢病毒转染和过继转移，改变CHI3L1在鼻黏膜上皮和巨噬细胞中的表达；发现CHI3L1对嗜酸粒细胞性CRSwNP黏膜炎症有促进作用；采用基因芯片技术筛选出CHI3L1下游重要分子AhR； 并在人体CRS组织中确认AhR主要表达于肥大细胞，采用体外肥大细胞培养技术，培养小鼠骨髓来源的肥大细胞，研究CHI3L1下游分子AhR对肥大细胞功能的影响，在体外研究CHI3L1及其下游分子对鼻-鼻窦黏膜炎症反应调控的精确分子机制；本研究项目揭示了CHI3L1在嗜酸粒细胞性CRSwNP黏膜炎症中的具体机制，深入认识了CHI3L1的功能，为发现治疗CRSwNP药物靶点打下基础。