Human bronchial epithelial cells exposed in vitro to cigarette smoke at the air-liquid interface resemble bronchial epithelium from human smokers

Abstract

Organotypic culture of human primary bronchial epithelial cells is a useful in vitro system to study normal biological processes and lung disease mechanisms, to develop new therapies, and to assess the biological perturbations induced by environmental pollutants. Herein, we investigate whether the perturbations induced by cigarette smoke (CS) and observed in the epithelium of smokers' airways are reproducible in this in vitro system (AIR-100 tissue), which has been shown to recapitulate most of the characteristics of the human bronchial epithelium. Human AIR-100 tissues were exposed to mainstream CS for 7, 14, 21, or 28 min at the air-liquid interface, and we investigated various biological endpoints [e.g., gene expression and microRNA profiles, matrix metalloproteinase 1 (MMP-1) release] at multiple postexposure time points (0.5, 2, 4, 24, 48 h). By performing a Gene Set Enrichment Analysis, we observed a significant enrichment of human smokers' bronchial epithelium gene signatures derived from different public transcriptomics datasets in CS-exposed AIR-100 tissue. Comparison of in vitro microRNA profiles with microRNA data from healthy smokers highlighted various highly translatable microRNAs associated with inflammation or with cell cycle processes that are known to be perturbed by CS in lung tissue. We also found a dose-dependent increase of MMP-1 release by AIR-100 tissue 48 h after CS exposure in agreement with the known effect of CS on this collagenase expression in smokers' tissues. In conclusion, a similar biological perturbation than the one observed in vivo in smokers' airway epithelium could be induced after a single CS exposure of a human organotypic bronchial epithelium-like tissue culture.

Reduced biological effect of e-cigarette aerosol compared to cigarette smoke evaluated in vitro using normalized nicotine dose and RNA-seq-based toxicogenomics

“…Cross-comparisons between transcriptomic studies from Beane et al, Chari et 6,[37][38][39][40] and the work presented here only show a 3-6% gene overlap. This lack of consensus can be attributed at least in part to the design of the studies with cells coming from either the upper or lower respiratory tract, different smoking doses and regimens.…”

Involvement of Toll-Like Receptor 2 and Epidermal Growth Factor Receptor Signaling in Epithelial Expression of Airway Remodeling Factors

“…MMP-1 was found to be highly elevated in patients with asthma, and was shown to be induced from airway epithelial cells by cigarette smoke extract, poly I:C, and respiratory syncytial virus (56)(57)(58). One of the main properties of MMP-1 is a proteolytic ability to degrade fibrillar collagen.…”

3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product

“…For each of the repetitions, three independent exposure runs were performed (each exposure run was conducted with 28 min continuous exposure to the smoke/aerosol). The duration of exposure was chosen based on the previously reported dose-dependent response of matrix metalloproteinase (MMP)1 secretion; the secretion of MMP1 was considered as a control for the responsiveness of organotypic airway cultures to CS (Mathis et al, 2013). Mathis et al reported that the highest concentration of secreted MMP1 was observed in a 3-D bronchial culture that closely mimic the in vivo environments.…”