An ex vivo model to induce early fibrosis-like changes in human precision-cut lung slices.
Idiopathic pulmonary fibrosis (IPF) is a devastating chronic interstitial lung disease (ILD) characterized by lung tissue scarring and high morbidity. Lung epithelial injury, myofibroblast activation, and deranged repair are believed to be key processes involved in disease onset and progression, but the exact molecular mechanisms behind IPF remain unclear. Several drugs have been shown to slow disease progression, but treatments that halt or reverse IPF progression have not been identified. Ex vivo models of human lung have been proposed for drug discovery, one of which is precision-cut lung slices (PCLS). Although PCLS production from IPF explants is possible, IPF explants are rare and typically represent end-stage disease. Here we present a novel model of early fibrosis-like changes in human PCLS derived from patients without ILD/IPF using a combination of profibrotic growth factors and signaling molecules (transforming growth factor-β, tumor necrosis factor-α, platelet-derived growth factor-AB, and lysophosphatidic acid). Fibrotic-like changes of PCLS were qualitatively analyzed by histology and immunofluorescence and quantitatively by water-soluble tetrazolium-1, RT-qPCR, Western blot analysis, and ELISA. PCLS remained viable after 5 days of treatment, and fibrotic gene expression (FN1,SERPINE1,COL1A1,CTGF,MMP7, andACTA2)increased as early as 24 h of treatment, with increases in protein levels at 48 h and increased deposition of extracellular matrix. Alveolar epithelium reprogramming was evident by decreases in surfactant protein C and loss ofHOPXIn summary, using human-derived PCLS, we established a novel ex vivo model that displays characteristics of early fibrosis and could be used to evaluate novel therapies and study early-stage IPF pathomechanisms.
Helmholtz Zentrum Munich, Comprehensive Pneumology Center, Member of the German Center for Lung Research, Munich, Germany.
Hani N
HN
Alsafadi
Helmholtz Zentrum Munich, Comprehensive Pneumology Center, Member of the German Center for Lung Research, Munich, Germany.
Claudia A
CA
Staab-Weijnitz
Helmholtz Zentrum Munich, Comprehensive Pneumology Center, Member of the German Center for Lung Research, Munich, Germany.
Mareike
M
Lehmann
Asklepios Fachkliniken München-Gauting Center of Thoracic Surgery, Gauting, Germany; and.
Michael
M
Lindner
Helmholtz Zentrum Munich, Comprehensive Pneumology Center, Member of the German Center for Lung Research, Munich, Germany.
Britta
B
Peschel
Helmholtz Zentrum Munich, Comprehensive Pneumology Center, Member of the German Center for Lung Research, Munich, Germany.|Division of Pulmonary Sciences and Critical Care Medicine Department of Medicine, University of Colorado Denver, Aurora, Colorado.
Melanie
0000-0001-9414-5128
M
Königshoff
Helmholtz Zentrum Munich, Comprehensive Pneumology Center, Member of the German Center for Lung Research, Munich, Germany; darcy.wagner@helmholtz-muenchen.de.
Darcy E
0000-0003-3794-1309
DE
Wagner
D000368:Aged; D056809:Alveolar Epithelial Cells / Q000378:metabolism / Q000473:pathology; D015415:Biomarkers / Q000378:metabolism; D024042:Collagen Type I / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D006801:Humans; D054990:Idiopathic Pulmonary Fibrosis / Q000473:pathology*; D018836:Inflammation Mediators / Q000378:metabolism; D008168:Lung / Q000473:pathology*; D008297:Male; D008954:Models, Biological; D014022:Tissue Survival; D015854:Up-Regulation
D015415:Biomarkers; D024042:Collagen Type I; D018836:Inflammation Mediators
disease model; ex vivo; fibrosis; idiopathic pulmonary fibrosis; precision-cut lung slices