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An ex vivo model to induce early fibrosis-like changes in human precision-cut lung slices

Alsafadi, Hani N.; Staab-Weijnitz, Claudia A.; Lehmann, Mareike; Lindner, Michael; Peschel, Britta; Königshoff, Melanie and Wagner, Darcy E. LU (2017) In American Journal of Physiology - Lung Cellular and Molecular Physiology 312(6). p.896-902
Abstract

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... (More)

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, and ACTA2) 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 of HOPX. In summary, using humanderived 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.

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author
publishing date
type
Contribution to journal
publication status
published
keywords
Disease model, Ex vivo, Fibrosis, Idiopathic pulmonary fibrosis, Precision-cut lung slices
in
American Journal of Physiology - Lung Cellular and Molecular Physiology
volume
312
issue
6
pages
896 - 902
publisher
American Physiological Society
external identifiers
  • scopus:85020229711
ISSN
1040-0605
DOI
10.1152/ajplung.00084.2017
language
English
LU publication?
no
id
77d1714a-ac9d-43dc-a270-60f77feeaeb8
date added to LUP
2017-08-15 14:33:37
date last changed
2017-09-03 05:26:06
@article{77d1714a-ac9d-43dc-a270-60f77feeaeb8,
  abstract     = {<p>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, and ACTA2) 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 of HOPX. In summary, using humanderived 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.</p>},
  author       = {Alsafadi, Hani N. and Staab-Weijnitz, Claudia A. and Lehmann, Mareike and Lindner, Michael and Peschel, Britta and Königshoff, Melanie and Wagner, Darcy E.},
  issn         = {1040-0605},
  keyword      = {Disease model,Ex vivo,Fibrosis,Idiopathic pulmonary fibrosis,Precision-cut lung slices},
  language     = {eng},
  number       = {6},
  pages        = {896--902},
  publisher    = {American Physiological Society},
  series       = {American Journal of Physiology - Lung Cellular and Molecular Physiology},
  title        = {An ex vivo model to induce early fibrosis-like changes in human precision-cut lung slices},
  url          = {http://dx.doi.org/10.1152/ajplung.00084.2017},
  volume       = {312},
  year         = {2017},
}