Chakraborty Ashesh, Giraldo-Arias Juliana, Merl-Pham Juliane, Dick Elisabeth, Mastalerz Michal, Zöller Marie, Marchi Hannah, Le Gleut Ronan, Hatz Rudolf A, Behr Jürgen, Hilgendorff Anne, Hauck Stefanie M, Staab-Weijnitz Claudia A
Institute of Lung Health and Immunity and Comprehensive Pneumology Center with the CPC-M bioArchive, Ludwig-Maximilians-Universität München and Helmholtz Zentrum München, Member of the German Center for Lung Research (DZL), 85764 Munich, Germany.
Metabolomics and Proteomics Core, Helmholtz Zentrum München, 85764 Neuherberg, Germany.
Int J Mol Sci. 2025 Aug 27;26(17):8324. doi: 10.3390/ijms26178324.
Cholesterol is an essential plasma membrane component, and altered cholesterol metabolism has been linked to cholesterol accumulation in the airways of COPD and cystic fibrosis patients. However, its role in airway epithelial differentiation is not well understood. Tandem mass spectrometry-based proteomic analysis of differentiating primary human bronchial epithelial cells (phBECs) revealed an overall inhibition of the cholesterol biosynthesis pathway. We hypothesized that excess cholesterol impairs the differentiation of phBECs into a fully functional bronchial epithelium. PhBECs were differentiated in the presence of 80 µM cholesterol for 21 days, the main airway cell type populations monitored using qRT-PCR and immunofluorescent stainings, and epithelial barrier integrity was analyzed via transepithelial electrical resistance measurements. Chronic cholesterol exposure led to a significant increase in CC10 secretory cells at the expense of ciliated cells. Pathway enrichment analysis suggested the tumor protein p53 as a master regulator of genes during normal differentiation of phBECs. Chronic cholesterol exposure drastically impaired the nuclear translocation of p53. Our findings suggest that this inhibition underlies the cholesterol-induced expansion of CC10 secretory cell populations at the expense of ciliated cells. In conclusion, we identify cholesterol as an important regulator of normal bronchial epithelial cell differentiation through inhibition of p53 nuclear translocation.
胆固醇是细胞膜的重要组成成分，而胆固醇代谢异常与慢性阻塞性肺疾病（COPD）和囊性纤维化患者气道内胆固醇蓄积有关。然而，其在气道上皮细胞分化中的作用尚不清楚。基于串联质谱的原代人支气管上皮细胞（phBECs）分化蛋白质组学分析显示胆固醇生物合成途径整体受到抑制。我们推测过量胆固醇会损害phBECs向功能完整的支气管上皮细胞分化。将phBECs在含有80µM胆固醇的条件下分化21天，使用qRT-PCR和免疫荧光染色监测主要气道细胞类型群体，并通过跨上皮电阻测量分析上皮屏障完整性。长期暴露于胆固醇导致CC10分泌细胞显著增加，而纤毛细胞数量减少。通路富集分析表明肿瘤蛋白p53是phBECs正常分化过程中基因的主要调节因子。长期暴露于胆固醇会严重损害p53的核转位。我们的研究结果表明，这种抑制作用是胆固醇诱导CC10分泌细胞群体增加并以纤毛细胞为代价的基础。总之，我们通过抑制p53核转位确定胆固醇是正常支气管上皮细胞分化的重要调节因子。
