Insights into epithelial-mesenchymal transition from cystic fibrosis rat models

• Published in: Journal of cystic fibrosis 2024-09
• Main Authors: Rout-Pitt, Nathan, Boog, Bernadette, McCarron, Alexandra, Reyne, Nicole, Parsons, David, Donnelley, Martin
• Format: Article
• Language: English
• Subjects: Airways; Cystic fibrosis; Epithelial mesenchymal transition; TGFβ1
• ISSN: 1569-1993; 1873-5010; 1873-5010
• DOI: 10.1016/j.jcf.2024.09.003

•Twist1 and Zeb1 gene expression is elevated in untreated Phe508del and KO cells.•Type 1 collagen is elevated in Phe508del and decreased in KO TGFβ1 treated cells.•ROCK inhibitor can normalise EMT changes in Phe508del, but not KO cells.•Further analysis of other mutation classes should be assessed. Molecular pathways contributing to Cystic Fibrosis pathogenesis remain poorly understood. Epithelial-mesenchymal transition (EMT) has been recently observed in CF lungs and certain CFTR mutation classes may be more susceptible than others. No investigations of EMT processes in CF animal models have been reported. The aim of this study was to assess the expression of EMT-related markers in Phe508del and knockout (CFTR-KO) rat lung tissue and tracheal-derived basal epithelial stem cells, to determine whether CFTR dysfunction can produce an EMT state. The expression of EMT-related markers in lung tissue and cultured tracheal basal epithelial stem cells from wildtype (WT), Phe508del, and CFTR-KO rats were assessed using qPCR and Western blots. Cell responses were evaluated in the presence of Rho-associated protein kinase (ROCK) inhibitor Y27632, which blocks EMT-pathways, or after treatment with TGFβ1 to stimulate EMT. Different gene expression profiles were observed between Phe508del and CFTR-KO rat models compared to wild type. There was lower expression of type 1 collagen in KO lungs and primary cell cultures, while Phe508del lungs and cells had higher expression, particularly when treated with TGFβ1. The addition of Y27632 rescued changes in EMT related genes in Phe508del cells but not in KO cells. Our findings show the first evidence of upregulated EMT pathways in the lungs and airway cells of any CF animal model. Differences in the regulation of the EMT genes and proteins in the Phe508del and CFTR-KO cells suggest that the signalling pathways underlying EMT are CFTR mutation dependent.