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Anti-fibrotic effects of Nintedanib and Pirfenidone on alveolar epithelial cell function in ex-vivo 3D-Lung Tissue Cultures
Anti-fibrotic effects of Nintedanib and Pirfenidone on alveolar epithelial cell function in ex-vivo 3D-Lung Tissue Cultures
Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disease of unknown cause with restricted therapeutic options. To date, the only approved drugs known to slow disease progression are Nintedanib and Pirfenidone. However, the life expectancy is still limited with or without treatment. Therefore, novel therapeutic options are needed. In this context, a more in- depth understanding of the mechanism of action of Nintedanib and Pirfenidone might help to identify novel therapeutic targets. Furthermore, the potential effect of these drugs on alveolar epithelial cells with being the key driver of the disease can be of particular interest. Therefore, in this study we aim to delineate the potential anti-fibrotic effects of both Nintedanib and Pirfenidone on alveolar epithelial Type II (AT II) cells using primary murine AT II cell culture as well as ex-vivo 3D-Lung Tissue Cultures (3D-LTCs). The Bleomycin-induced lung fibrosis model in mice and the Fibrotic Cocktail (FC), consisting of multiple pro-fibrotic growth factors, in human 3D-LTCs were mainly utilized for this study. Firstly, we demonstrated pro-fibrotic hallmarks to be recapitulated in both murine and human fibrotic ex-vivo 3D-LTCs during culture duration. Nintedanib and Pirfenidone subsequently inhibited fibrotic marker gene and protein expression in murine ex-vivo 3D-LTCs and primary murine AT II cell culture. We showed Nintedanib, but not Pirfenidone, to induce AT II cell marker expression, in particular Surfactant Protein-C (SP-C), in both primary murine AT II cell culture and ex-vivo 3D-LTCs. Our data further indicated that this effect is partly mediated by Nkx2.1 and Hopx, two transcription factors being crucial for AT II transdifferentiation, both being upregulated upon Nintedanib treatment. Interestingly, we were able to recapitulate the effect of Nintedanib on AT II cell marker, but not on fibrotic marker expression in human ex-vivo 3D-LTCs. In summary, we show murine and human ex-vivo 3D-LTCs to be a feasible tool for preclinical drug testing. We further showed differential effects of Nintedanib and Pirfenidone on epithelial cell marker expression possibly contributing to their anti-fibrotic action.
IPF, Nintedanib, Pirfenidone, alveolar epithelial cells, ex-vivo 3D-Lung Tissue Cultures
Buhl, Lara
2021
English
Universitätsbibliothek der Ludwig-Maximilians-Universität München
Buhl, Lara (2021): Anti-fibrotic effects of Nintedanib and Pirfenidone on alveolar epithelial cell function in ex-vivo 3D-Lung Tissue Cultures. Dissertation, LMU München: Faculty of Medicine
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Abstract

Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disease of unknown cause with restricted therapeutic options. To date, the only approved drugs known to slow disease progression are Nintedanib and Pirfenidone. However, the life expectancy is still limited with or without treatment. Therefore, novel therapeutic options are needed. In this context, a more in- depth understanding of the mechanism of action of Nintedanib and Pirfenidone might help to identify novel therapeutic targets. Furthermore, the potential effect of these drugs on alveolar epithelial cells with being the key driver of the disease can be of particular interest. Therefore, in this study we aim to delineate the potential anti-fibrotic effects of both Nintedanib and Pirfenidone on alveolar epithelial Type II (AT II) cells using primary murine AT II cell culture as well as ex-vivo 3D-Lung Tissue Cultures (3D-LTCs). The Bleomycin-induced lung fibrosis model in mice and the Fibrotic Cocktail (FC), consisting of multiple pro-fibrotic growth factors, in human 3D-LTCs were mainly utilized for this study. Firstly, we demonstrated pro-fibrotic hallmarks to be recapitulated in both murine and human fibrotic ex-vivo 3D-LTCs during culture duration. Nintedanib and Pirfenidone subsequently inhibited fibrotic marker gene and protein expression in murine ex-vivo 3D-LTCs and primary murine AT II cell culture. We showed Nintedanib, but not Pirfenidone, to induce AT II cell marker expression, in particular Surfactant Protein-C (SP-C), in both primary murine AT II cell culture and ex-vivo 3D-LTCs. Our data further indicated that this effect is partly mediated by Nkx2.1 and Hopx, two transcription factors being crucial for AT II transdifferentiation, both being upregulated upon Nintedanib treatment. Interestingly, we were able to recapitulate the effect of Nintedanib on AT II cell marker, but not on fibrotic marker expression in human ex-vivo 3D-LTCs. In summary, we show murine and human ex-vivo 3D-LTCs to be a feasible tool for preclinical drug testing. We further showed differential effects of Nintedanib and Pirfenidone on epithelial cell marker expression possibly contributing to their anti-fibrotic action.