Abstract 4134
Background
Numerous clinical trials are currently evaluating anti-transforming growth factor beta (TGF-β) therapy for treating lung cancer patients. However, interrogating stromal reactivity, and enhancing the mechanistic understanding of desmoplasia in relation to TGF-β signaling represents unmet medical needs and may allow for discovery of novel TGF-β associated biomarkers for use in the clinical setting. Cancer associated fibroblasts are major contributors to the desmoplastic reaction (ECM deposition) upon TGF-β stimulation. Applying the “Scar-in-a-jar” (SiaJ) model, we evaluated the impact of TGF-β, and inhibitors, on lung fibroblasts’ expression of different collagens.
Methods
Primary human healthy lung fibroblasts were cultured for up to 15 days in the presence of ficoll and TGF-β, with or without addition of 1nM-10µM ALK-5/type I TGF-β receptor kinase inhibitor (iTGFβ. ELISAs quantified pro-peptides from type I (PINP), type III (PRO-C3) and type VI (PRO-C6) collagen in cell supernatant as surrogate measures of the TGF-β induced ECM deposition. Cytotoxicity (lactate dehydrogenase (LDH) release) and metabolic activity (AlamarBlue) were evaluated.
Results
Stimulating lung fibroblasts with TGF-β induced PINP, PRO-C3 and PRO-C6 increase up to 8-fold compared to TGF-β (p < 0.001). iTGFβ dose-dependently reduced the PINP, PRO-C3 and PRO-C6 increase induced by TGF-β. No cytotoxicity could be detected. The metabolic activity was decreased at 1uM iTGFβ.
Conclusions
The SiaJ model can be used to evaluate the impact of TGF-β, and inhibitors, on lung fibroblasts’ viability and expression of different collagens. The findings suggest that SiaJ together with PINP, PRO-C3 and PRO-C6 can be used as a translational drug screening tool for interrogating the effect of anti-TGF-β therapy on fibroblast activity and the desmoplastic reaction.
Clinical trial identification
Legal entity responsible for the study
Nordic Bioscience A/S.
Funding
Nordic Bioscience A/S.
Editorial Acknowledgement
Disclosure
N.I. Nissen, N. Willumsen, N. Gudmann, S. Rønnow, J. Sand: Employee: Nordic Bioscience A/S. M. Karsdal, D. Leeming: Employee and stocks: Nordic Bioscience A/S.