Differential effects of Nintedanib and Pirfenidone on lung alveolar epithelial cell function in ex vivo murine and human lung tissue cultures of pulmonary fibrosis

Mareike Lehmann,Lara Buhl,Jürgen Behr,Kathrin Mutze,Melanie Königshoff,Sarah Hermann,Stephan Klee,Anne Hilgendorff,Darcy E Wagner,Michael Lindner,Chiharu Ota,Hani N Alsafadi

doi:10.1186/s12931-018-0876-y

Abstract

BackgroundIdiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease. Repetitive injury and reprogramming of the lung epithelium are thought to be critical drivers of disease progression, contributing to fibroblast activation, extracellular matrix remodeling, and subsequently loss of lung architecture and function. To date, Pirfenidone and Nintedanib are the only approved drugs known to decelerate disease progression, however, if and how these drugs affect lung epithelial cell function, remains largely unexplored.MethodsWe treated murine and human 3D ex vivo lung tissue cultures (3D-LTCs; generated from precision cut lung slices (PCLS)) as well as primary murine alveolar epithelial type II (pmATII) cells with Pirfenidone or Nintedanib. Murine 3D-LTCs or pmATII cells were derived from the bleomycin model of fibrosis. Early fibrotic changes were induced in human 3D-LTCs by a mixture of profibrotic factors. Epithelial and mesenchymal cell function was determined by qPCR, Western blotting, Immunofluorescent staining, and ELISA.ResultsLow μM concentrations of Nintedanib (1 μM) and mM concentrations of Pirfenidone (2.5 mM) reduced fibrotic gene expression including Collagen 1a1 and Fibronectin in murine and human 3D-LTCs as well as pmATII cells. Notably, Nintedanib stabilized expression of distal lung epithelial cell markers, especially Surfactant Protein C in pmATII cells as well as in murine and human 3D-LTCs.ConclusionsPirfenidone and Nintedanib exhibit distinct effects on murine and human epithelial cells, which might contribute to their anti-fibrotic action. Human 3D-LTCs represent a valuable tool to assess anti-fibrotic mechanisms of potential drugs for the treatment of IPF patients.

Highlights

Idiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease
Nintedanib (BIBF-1120) is a multi-tyrosine kinase inhibitor and is known to inhibit the receptor kinases of platelet-derived growth factor (PDGF), fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF), which are all thought to play an important role in the pathogenesis of IPF [16]
As our ultimate goal was to test the effect of the compounds in an ex vivo model of human pulmonary fibrosis, we sought to establish the feasibility of this approach by using murine ex vivo models

Summary

Introduction

Idiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease. Repetitive injury and reprogramming of the lung epithelium are thought to be critical drivers of disease progression, contributing to fibroblast activation, extracellular matrix remodeling, and subsequently loss of lung architecture and function. Alveolar epithelial type (AT) II cells have been described to undergo significant phenotypic and functional changes upon fibrotic lung injury [4, 5], including endoplasmatic reticulum stress [6], mitochondrial dysfunction [7], or senescence [8], these cells have been reported to release a variety of profibrotic mediators, such as Transforming growth factor (TGF)-β [9], IL-1β [10], sphingosine 1-phosphate (S1P) [11], or WNT ligands [12, 13] These studies support the notion that targeting the dysfunctional epithelium might be a promising therapeutic strategy for the treatment of IPF. Pirfenidone has anti-oxidant, anti-fibrotic and anti-inflammatory properties as shown in several in vitro and in vivo studies [22, 23]

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