Manon Barthe (1) , Laurie Perdigon (1) , Hanan Osman-Ponchet (1) , Agnès Choppin (2) , Franck Chiapini (2)
Pulmonary fibrosis is a progressive, restrictive lung disease characterized by scarring and stiffening of lung tissue, ultimately leading to respiratory failure. This condition is driven by the excessive proliferation and migration of fibroblasts, specialized cells residing within the lung. These activated fibroblasts deposit excessive amounts of extracellular matrix (ECM) proteins, fundamentally disrupting the lung’s architecture and hindering its ability to function properly. Therefore, targeting fibroblast proliferation and associated ECM deposition represents a logical therapeutic strategy for both delaying disease onset and, potentially, reversal of disease pathology. In this study, we utilized primary human pulmonary fibroblasts to develop a functional screening assay for identifying novel anti-fibrotic drugs. Primary human pulmonary fibroblasts (HPFs) were cultured in Ibidi® two-well inserts and treated with either nintedanib (a tyrosine kinase inhibitor) or pirfenidone (an anti-inflammatory drug), both FDA-approved treatments for pulmonary fibrosis. To assess the impact of these drugs on cell migration, the wounded area of the inserts was imaged using a CytoSMART Omni brightfield live-cell imager, capturing time-lapse images every hour for up to 24 hours. In parallel, separate cultures of HPFs were treated with either TGF-β alone or in combination with nintedanib. Untreated cells served as control. After 24 hours, mRNA expression of fibrosis markers in these cells was measured using RT-qPCR. Our findings revealed that pirfenidone had minimal effect on HPF migration, while nintedanib significantly reduced their migratory capacity. Furthermore, treatment with TGF-β, a known fibrosis inducer, led to increased expression of fibrosis markers, particularly LRRC15, COL1A1, and FN1. Importantly, nintedanib treatment effectively countered this effect by decreasing the expression of TGF-β-induced fibrosis markers. In conclusion, this study established a functional screening assay for identifying potential anti-fibrotic drugs using primary human pulmonary fibroblasts. This assay combines cell migration and gene expression analysis is a simple, versatile, and cost-effective method for the screening of novel anti-fibrotic therapeutic treatments targeting pulmonary fibrosis.
