人气道基底干细胞向肺泡上皮细胞转化的wnt信号机制研究

Idiopathic Pulmonary Fibrosis (IPF) is a kind of interstitial lung disease and no effective treatment currently. IPF is associated with dysfunction of stem cells in alveoli. Airway basal cells（BCs）are ideal therapeutic stem cells which can effect as supplement of alveolar stem cells because of their strong plastically. Our previous work has proved that BCs have good effect on IPF patients, but how to accurately induce BCs transforming to alveolar epithelial cells is worth to study. Wnt signaling pathway promotes alveolar development and alveolar epithelial cells transformation in embryonic period, but if it can induce adult BCs transform to alveolar epithelial cells remains unknown. We plan to inhibition/active Wnt signaling pathway by knockdown/overexpression β-catenin, which is a key downstream gene in Wnt signaling in BCs in advance, followed by differentiation experiments in vitro (air-liquid interface system and 3D differentiation system) and in vivo (in mouse model of pulmonary fibrosis) to compare BCs transformation in active/inhibition Wnt signaling BC by techniques such as IHC、animal CT and so on. Next we plan to explore mechanisms of Wnt signaling pathway in BCs to alveolar epithelial cells transformation by techniques such as qPCR、western blot and so on. We hope our experiments could reveal the role of Wnt signaling pathway in regulating of BCs’ transformation, and provide new approach to improve IPF treatment.

特发性肺纤维化（IPF）是常见肺间质疾病，目前无有效疗法。IPF发病与肺泡干细胞功能低下有关。气道基底细胞（BCs）可塑性强，可弥补肺泡干细胞不足，是IPF治疗理想干细胞。本团队前期研究已证实自体BCs移植对IPF有一定治疗作用，但如何精准调控BCs向肺泡上皮细胞分化、提高治疗效率，值得深入研究。Wnt信号通路促进肺泡发育及肺泡上皮再生，其是否可调控成体BCs向肺泡上皮细胞定向分化尚无研究。本课题拟通过沉默/过表达β-catenin基因对BCs内Wnt信号进行抑制/激活预处理，随后在气-液交界培养、3D培养两种体外分化及小鼠肺纤维化模型体内分化条件下，通过免疫组化、影像学等方法比较不同Wnt信号水平对BCs分化命运的影响；并通过qPCR、WB等技术对机制进行探索，从而明确Wnt信号通路在调控BCs上皮细胞方向分化中的具体作用，为提高IPF治疗效率提供新思路。

特发性肺纤维化（IPF）是常见肺间质疾病，目前无有效疗法。IPF发病与肺泡干细胞功能低下有关。气道基底细胞（BCs）可塑性强，可弥补肺泡干细胞不足，是IPF治疗理想干细胞。本团队前期研究已证实自体BCs移植对IPF有一定治疗作用，但如何精准调控BCs向肺泡上皮细胞分化、提高治疗效率，值得深入研究。Wnt信号通路促进肺泡发育及肺泡上皮再生，其是否可调控成体BCs向肺泡上皮细胞定向分化尚无研究。本课题通过比较激活不同Wnt信号通路（经典通路、非经典通路）对BCs增值生长、分化命运的影响，并通过RNA-seq、WB等技术对机制进行探索，从而明确Wnt信号通路在调控BCs上皮细胞方向分化中的具体作用，为提高IPF治疗效率提供新思路。本课题实验结果表明，激活Wnt经典通路及非经典通路均可促进BCs的克隆增殖及上皮分化趋势，Wnt5A激活的非经典途径略优于Wnt3A激活的经典途径，其机制虽尚未完全明确，但为BCs的定向分化研究提供了支持。