Epithelial 3D-spheroids as a tool to study air pollutant-induced lung pathology

Cigarette smoke (CS) and air pollutants (AP) activate pathological processes in bronchial epithelial cells resulting in lung function decline which severely impacts human health. Knowledge about the molecular mechanism(s) by which CS and AP induce pathology is limited. Our previous studies in 2D cultures of human bronchial epithelial (BEAS-2B) cells showed that CS exposure activates transforming growth factor-beta 1 (TGF-beta 1) release and signaling. Furthermore, CS exposure reduced the expression of E-cadherin, which was prevented by applying a TGE-beta 1 neutralizing antibody. Exposure of BEAS-2B cells cultured in 2D to diesel exhaust particles (DEP) increased TGE-beta 1 protein expression and reduced the expression of epithelial cell markers, whereas mesenchymal markers are upregulated. Conventional 2D cell culture may, however, not fully reflect the physiology of bronchial epithelial cells in vivo. To simulate the in vivo situation more closely we cultured the bronchial epithelial cells in a 3D environment in the current study. Treatment of epithelial spheroids with TGE-fi resulted in reduced E-cadherin and increased collagen I expression, indicating the activation of epithelial-to-mesenchymal transition (EMT). Similarly, exposure of spheroids to DEP induced and EMT-like phenotype. Collectively, our data indicate AP induces an EMT-like phenotype of BEAS-2B cells in 3D spheroid cultures. This opens new avenues for drug development for the treatment of lung diseases induced by AP. The 3D spheroid cell culture is a novel, innovative and physiologically relevant model for culturing a variety of cells. It is a versatile tool for both high-throughput studies and for identifying molecular mechanisms involved in bronchial epithelial cell (patho)physiology.

Automated summary

Cigarette smoke and air pollutants can cause pathological changes in bronchial epithelial cells, leading to lung function decline. Previous studies have shown that cigarette smoke exposure activates the release and signaling of TGF-beta 1, and reduces the expression of E-cadherin in 2D cell cultures. This study cultured bronchial epithelial cells in a 3D environment and found that cigarette smoke and air pollutants induce an epithelial-to-mesenchymal transition phenotype.