Evaluation of Pirfenidone and Nintedanib in a Human Lung Model of Fibrogenesis

Abstract

Introduction: Idiopathic pulmonary fibrosis (IPF) is a progressive, fatal lung disease with a poor prognosis and increasing incidence. Pirfenidone and nintedanib are the only approved treatments for IPF but have limited efficacy and their mechanisms of action are poorly understood. Here we have examined the effects of pirfenidone and nintedanib in a human model of lung fibrogenesis, and compared these with the putative anti-fibrotic compounds Lipoxin A4 (LXA4), and senicapoc, a KCa3.1 ion channel blocker.Methods: Early fibrosis was induced in cultured human lung parenchyma using TGFβ1 for 7 days, ± pirfenidone, nintedanib, or LXA4. Pro-fibrotic responses were examined by RT-PCR, immunohistochemistry and soluble collagen secretion.Results: Thirty six out of eighty four IPF and fibrosis-associated genes tested were significantly upregulated by TGFβ1 in human lung parenchyma with a ≥0.5 log2FC (n = 32). Nintedanib (n = 13) reduced the mRNA expression of 14 fibrosis-associated genes including MMPs (MMP1,−4,−13,−14), integrin α2, CXCR4 and PDGFB, but upregulated α-smooth muscle actin (αSMA). Pirfenidone only reduced mRNA expression for MMP3 and −13. Senicapoc (n = 11) previously attenuated the expression of 28 fibrosis-associated genes, including αSMA, several growth factors, collagen type III, and αV/β6 integrins. Pirfenidone and nintedanib significantly inhibited TGFβ1-induced fibroblast proliferation within the tissue, but unlike senicapoc, neither pirfenidone nor nintedanib prevented increases in tissue αSMA expression. LXA4 was ineffective.Conclusions: Pirfenidone and nintedanib demonstrate modest anti-fibrotic effects and provide a benchmark for anti-fibrotic activity of new drugs in human lung tissue. Based on these data, we predict that the KCa3.1 blocker senicapoc will show greater benefit than either of these licensed drugs in IPF.

Effects of anti-fibrotic standard of care drugs on senescent human lung fibroblasts

“…We observed an increase of α-SMA, collagen-1, and fibronectin 1 after the treatment with pirfenidone, suggesting that although evidence shows its anti-fibrotic effect in proliferative fibroblasts (COLLINS; RAGHU, 2019), the same effect is not perceived in senescent fibroblasts. In agreement with our findings, Roach et al (2021) and collaborators found an increased expression of α-SMA, deposition of collagen 1 and collagen-3, and secretion of soluble collagen after treating fibroblasts with nintedanib and pirfenidone (ROACH; CASTELLS; DIXON; MASON et al ., 2021).…”

Nintedanib regulates intestinal smooth muscle hyperplasia and phenotype in vitro and in TNBS colitis in vivo

“…These drugs have been studied in lung fibrosis, where both pulmonary smooth muscle and fibroblasts participate (e.g., Ref. 29 ), and so we also examined their effect on fibroblasts, using the 3T3 fibroblast cell line. Again, FCS-induced growth of 3T3 cells showed a far greater sensitivity to NIN (Fig.…”

Matrix Metalloproteinases and Their Inhibitors in Pulmonary Fibrosis: EMMPRIN/CD147 Comes into Play

“…These observations reveal that IL-1β may reduce local tissue rigidity and provide innate antifibrotic protection that could be important during the early stages of fibrotic processes and lung repair [24]. Other studies have demonstrated that the pan-RTK (receptor tyrosine kinase) inhibitor nintedanib can reduce the mRNA expression for mmp1, 4, 13, and 14 and pirfenidone is able to reduce mRNA expression for mmp3 and 13 [100]. In both cell culture and mouse models of large cell lung carcinoma cell lines (LCC), the overexpression of mmp1 has been described to be necessary for the induction of fibroblast senescence and consequent tumor promotion [101].…”

“…On the contrary, mmp3-null mice were protected from bleomycin-induced lung fibrosis [29]. Recent studies have described that pirfenidone, a drug approved for IPF treatment reduces mRNA expression of mmp3 in a profibrotic model of human lung fibroblasts stimulated by TGF-β1 [100]. In conclusion, it seems to be clear that MMP-3 has an important profibrotic function in the IPF context.…”