Ferret model of bleomycin-induced lung injury shares features of human idiopathic pulmonary fibrosis

Wu, S.; Driver, I.; Luo, M.; Miyazaki, H.; Shambhu, S.; Popov, D.; Yang, L.; Wang, J.; Ma, J.; Guo, J.; Atha, J.; Tata, A.; Hoffman, E.; Wang, Y.; Tata, P.; Jensen, M. B.; Engelhardt, J. F.; Sontake, V.; Liu, X.

doi:10.1101/2025.05.08.652970

bioRxiv · 10.1101/2025.05.08.652970

Ferret model of bleomycin-induced lung injury shares features of human idiopathic pulmonary fibrosis

Wu, S.·Driver, I.·Luo, M.·Miyazaki, H.·Shambhu, S.·Popov, D.·Yang, L.·Wang, J.·Ma, J.·Guo, J.·Atha, J.·Tata, A.·Hoffman, E.·Wang, Y.·Tata, P.·Jensen, M. B.·Engelhardt, J. F.·Sontake, V.·Liu, X.

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Abstract

Idiopathic pulmonary fibrosis (IPF) is a debilitating lung disease with limited therapeutic options. The development of effective therapies has been hindered by the lack of models that recapitulate key features of human disease. Here we report a bleomycin-induced ferret PF model characterized by an irreversible decrease in pulmonary compliance and an increase of opacification, accompanied by "honeycomb cyst-like" structures and "proximalization" of distal lung epithelium. Cellular and molecular analysis by single-nucleus RNA sequencing revealed a significant shift in distal lung epithelium towards proximal epithelial phenotype. Importantly, a histopathological pattern of bronchiolization encompassing divergent atypical epithelial cells and KRT17+/TP63+/KRT5low "basaloid-like" cells was present in the distal fibrotic lung lesions. Trajectory analysis revealed AT2 cells transition through multiple cell-states in bleomycin injured ferret lungs, particularly AT2 to KRT8high/KRT7low/SOX4+ and eventually to KRT8high/KRT7high/SFN+/TP63+/KRT5low "basaloid-like" cells. Further, immunofluorescence analyses demonstrated KRT7 and KRT8 populations reside overlaying the ACTA2-positive myofibroblasts in fibrotic foci, implying their pro-fibrogenic activity similar to human IPF lungs. Collectively, our results provide evidence that bleomycin-induced lung injury in ferrets recapitulates pathophysiological, cellular, and molecular features of human IPF, suggesting that they may be a reliable model for understanding mechanisms of IPF pathogenesis and for testing therapeutic strategies for treatment of IPF. Take Home MessageBleomycin-induced acute lung injury in the ferret recapitulates pathophysiological, cellular, and molecular features of human IPF; thus the ferret may be a reliable species for studying mechanisms of IPF pathogenesis and testing therapeutic strategies.

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Wu, S., Driver, I., Luo, M., Miyazaki, H., Shambhu, S., Popov, D., Yang, L., Wang, J., Ma, J., Guo, J., Atha, J., Tata, A., Hoffman, E., Wang, Y., Tata, P., Jensen, M. B., Engelhardt, J. F., Sontake, V., Liu, X.. 2025-05-14. Ferret model of bleomycin-induced lung injury shares features of human idiopathic pulmonary fibrosis. https://doi.org/10.1101/2025.05.08.652970

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