Abstract
The differentiated human airway epithelium consists of different cell types forming a polarized and pseudostratified epithelium. This is dramatically altered in chronic obstructive pulmonary disease (COPD), characterized by basal and goblet cell hyperplasia, and squamous cell metaplasia. The effect of cigarette smoke on human bronchial epithelial cell (HBEC) differentiation remains to be elucidated. We analysed whether cigarette smoke extract (CSE) affected primary (p)HBEC differentiation and function. pHBEC were differentiated at the air-liquid interface (ALI) and differentiation was quantified after 7, 14, 21, or 28 days by assessing acetylated tubulin, CC10, or MUC5AC for ciliated, Clara, or goblet cells, respectively. Exposure of differentiating pHBEC to CSE impaired epithelial barrier formation, as assessed by resistance measurements (TEER). Importantly, CSE exposure significantly reduced the number of ciliated cells, while it increased the number of Clara and goblet cells. CSE-dependent cell number changes were reflected by a reduction of acetylated tubulin levels, an increased expression of the basal cell marker KRT14, and increased secretion of CC10, but not by changes in transcript levels of CC10, MUC5AC, or FOXJ1. Our data demonstrate that cigarette smoke specifically alters the cellular composition of the airway epithelium by affecting basal cell differentiation in a post-transcriptional manner.
Highlights
Correspondence and requests for materials should be addressed to Cigarette smoke alters primary human bronchial epithelial cell differentiation at the air-liquid interface
We analysed whether cigarette smoke extract (CSE) affected primary (p)human bronchial epithelial cell (HBEC) differentiation and function. primary human bronchial epithelial cells (pHBECs) were differentiated at the air-liquid interface (ALI) and differentiation was quantified after 7, 14, 21, or 28 days by assessing acetylated tubulin, cell-specific 10 kDa protein (CC10), or MUC5AC for ciliated, Clara, or goblet cells, respectively
In order to examine the effect of cigarette smoke extract (CSE) on basal cell differentiation at the air-liquid interface (ALI), cells were exposed to CSE during the entire differentiation process and analysis was performed after 7, 14, 21, and 28 days of differentiation (Figure 1a)
Summary
Correspondence and requests for materials should be addressed to Cigarette smoke alters primary human bronchial epithelial cell differentiation at the air-liquid interface. The differentiated human airway epithelium consists of different cell types forming a polarized and pseudostratified epithelium This is dramatically altered in chronic obstructive pulmonary disease (COPD), characterized by basal and goblet cell hyperplasia, and squamous cell metaplasia. We and others reported recently that acute exposure to cigarette smoke extract (CSE) impairs barrier function and tight junction organization in vitro, using primary human bronchial epithelial cells (pHBECs) differentiated at the air-liquid interface (ALI)[24,25]. These changes in combination with mucus overproduction and decreased mucociliary clearance promote pathogen colonization and the development of smoking-associated lung disorders, such as COPD and lung cancer. Whether www.nature.com/scientificreports the smoking-associated changes in the airway epithelium are a detrimental effect of cigarette smoke on already differentiated cells, or whether continuous cigarette smoke exposure affects basal/intermediate cell differentiation
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