Chakraborty Ashesh, Zöller Marie, Sardogan Aydan, Klotz Markus, Mastalerz Michal, Marchi Hannah, Meixner Raphael, Hatz Rudolf A, Behr Jürgen, Hilgendorff Anne, Nakayama Misako, Staab-Weijnitz Claudia A
Helmholtz Center Munich German Research Center for Environmental Health, Institute for Lung Health and Immunity and Comprehensive Pneumology Center, Neuherberg, Bayern, Germany.
Helmholtz Center Munich German Research Center for Environmental Health, Institute of Regenerative Biology and Medicine (IRBM), Neuherberg, BY, Germany.
Am J Respir Cell Mol Biol. 2025 Apr 16. doi: 10.1165/rcmb.2024-0117OC.
The human airway epithelium is a primary site of toxicant exposure and crucial in the pathogenesis of acute and chronic lung disease (CLD). In CLD, the airway epithelium is frequently altered and distorted, and its restoration is desirable. The mechanisms underlying human aberrant epithelial regeneration, however, are poorly understood. Importantly, our knowledge about airway epithelial injury and regeneration largely stems from mouse models, yet airways differ considerably between mice and humans. We hypothesized that treatment of differentiated primary human bronchial epithelial cells (phBECs, or HBEC) with polidocanol or naphthalene would allow for studying mechanisms of human airway epithelial injury and regeneration. Injury of differentiated phBECs with 0.04%, but not 0.1% PDOC, resulted in full restoration of a functional epithelium and epithelial barrier integrity as monitored by qRT-PCR analysis, immunofluorescence stainings, and transepithelial electrical resistance measurements. Regeneration was associated with a transient but not parallel increase of p21+ and KRT17+ cells. Providing proof-of-concept, DAPT, an inhibitor of Notch signaling, blunted the restoration of secretory cell types post 0.04% PDOC injury. Differentiation of phBECs in presence of cigarette smoke extract (CSE) or ethanol as first hit significantly impaired the regeneration capacity of phBECs. While naphthalene is known to specifically induce club cell depletion in mouse airways, it failed to do so in phBECs. In conclusion, using fully differentiated phBECs treated with PDOC, we successfully established and thoroughly characterized a human system that will facilitate studies of mechanisms involved in susceptibility to injury as well as human airway repair and regeneration.
人类气道上皮是毒物暴露的主要部位，在急性和慢性肺部疾病（CLD）的发病机制中起关键作用。在CLD中，气道上皮经常发生改变和扭曲，其修复是理想的。然而，人类异常上皮再生的潜在机制尚不清楚。重要的是，我们对气道上皮损伤和再生的了解很大程度上源于小鼠模型，但小鼠和人类的气道有很大差异。我们假设用聚多卡醇或萘处理分化的原代人支气管上皮细胞（phBECs，或HBEC）将有助于研究人类气道上皮损伤和再生的机制。用0.04%而非0.1%的聚多卡醇损伤分化的phBECs，通过qRT-PCR分析、免疫荧光染色和跨上皮电阻测量监测，导致功能性上皮和上皮屏障完整性完全恢复。再生与p21+和KRT17+细胞的短暂但不平行增加有关。作为概念验证，Notch信号抑制剂DAPT抑制了0.04%聚多卡醇损伤后分泌细胞类型的恢复。在香烟烟雾提取物（CSE）或乙醇存在下作为首次打击使phBECs分化，显著损害了phBECs的再生能力。虽然已知萘能特异性诱导小鼠气道中的俱乐部细胞耗竭，但它在phBECs中未能做到这一点。总之，使用经聚多卡醇处理的完全分化的phBECs，我们成功建立并全面表征了一个人类系统，该系统将有助于研究与损伤易感性以及人类气道修复和再生相关的机制。
