Molecular characterization of a precision-cut rat lung slice model for the evaluation of antifibrotic drugs

• Published in: American journal of physiology. Lung cellular and molecular physiology Vol. 316; no. 2; pp. L348 - L357
• Main Authors: Huang, Xinqiang, Li, Li, Ammar, Ron, Zhang, Yan, Wang, Yihe, Ravi, Kandasamy, Thompson, John, Jarai, Gabor
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 01.02.2019
• Subjects: Animal models; Animals; Attenuation; Biomarkers; Biomarkers - metabolism; Cell adhesion; Cell adhesion & migration; Clinical trials; Collagen (type I); Collagen Type I - drug effects; Collagen Type I - metabolism; Drug development; Drugs; Effectiveness; Evaluation; Extracellular matrix; Extracellular Matrix - drug effects; Extracellular Matrix - metabolism; Fibrosis; Fibrosis - drug therapy; Gene expression; Gene sequencing; Genes; Growth factors; Hyaluronic acid; Immune response; Immunosuppressive agents; Indoles - pharmacology; Integrins; Lung - drug effects; Lung diseases; Medical research; Mesenchyme; Procollagen; Proteins; Pulmonary fibrosis; Pyridones - pharmacology; Ribonucleic acid; RNA; Signal transduction; Transforming growth factor; Transforming growth factor-b1; model; lung slice; pulmonary fibrosis
• ISSN: 1040-0605; 1522-1504
• DOI: 10.1152/ajplung.00339.2018

The translation of novel pulmonary fibrosis therapies from preclinical models into the clinic represents a major challenge demonstrated by the high attrition rate of compounds that showed efficacy in preclinical models but demonstrated no significant beneficial effects in clinical trials. A precision-cut lung tissue slice (PCLS) contains all major cell types of the lung and preserves the original cell-cell and cell-matrix contacts. It represents a promising ex vivo model to study pulmonary fibrosis. In this study, using RNA sequencing, we demonstrated that transforming growth factor-β1 (TGFβ1) induced robust fibrotic responses in the rat PCLS model, as it changed the expression of genes functionally related to extracellular matrix remodeling, cell adhesion, epithelial-to-mesenchymal transition, and various immune responses. Nintedanib, pirfenidone, and sorafenib each reversed a subset of genes modulated by TGFβ1, and of those genes we identified 229 whose expression was reversed by all three drugs. These genes define a molecular signature characterizing many aspects of pulmonary fibrosis pathology and its attenuation in the rat PCLS fibrosis model. A panel of 12 genes and three secreted biomarkers, including procollagen I, hyaluronic acid, and WNT1-inducible signaling pathway protein 1 were validated as efficacy end points for the evaluation of antifibrotic activity of experimental compounds. Finally, we showed that blockade of α -integrins suppressed TGFβ1-induced fibrotic responses in the rat PCLS fibrosis model. Overall, our results suggest that the TGFβ1-induced rat PCLS fibrosis model may represent a valuable system for target validation and to determine the efficacy of experimental compounds.