Luciferase expression is driven by the promoter of fibroblast activation protein-α in murine pulmonary fibrosis

Biotechnol Lett. 2015 Sep;37(9):1757-63. doi: 10.1007/s10529-015-1855-8. Epub 2015 May 21.

Abstract

Objectives: We used optical imaging of live animals and transgenic technology to develop a pulmonary fibrosis model in mice that can non-invasively and in real-time trace the pulmonary fibrosis process.

Results: Fibroblast activation protein-α (FAPα) is selectively expressed in fibrotic foci of human pulmonary fibrosis. It is not expressed in normal tissue. We confirmed that FAPα is upregulated in fibroblasts of murine pulmonary fibrosis. Moreover, TGF-β1, a central pathological mediator of fibrotic diseases, could promote FAPα expression in mouse embryonic fibroblasts. Luciferase reporter assays showed that 5.4 kb FAPα promoter response activities to TGF-β1 was stronger than of the 2.1 kb promoter. We generated a transgenic mouse line expressing firefly luciferase under the control of the 5.4 kb FAPα gene promoter (FAPα-p-luc). After experimentally inducing murine pulmonary fibrosis, there luminescence appeared in the chests and excised lungs of FAPα-p-luc mice. The intensity of luminescence became stronger with the exacerbation of pulmonary fibrosis.

Conclusion: Fluorescence intensity reflects the degree of pulmonary fibrosis in FAPα-p-luc mice. and this mouse model may be used to investigate molecular mechanisms and drug screening of pulmonary fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bleomycin / pharmacology
  • Cells, Cultured
  • Disease Models, Animal
  • Endopeptidases
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Gelatinases / genetics*
  • Gelatinases / metabolism
  • Humans
  • Luciferases, Firefly / genetics
  • Luciferases, Firefly / metabolism*
  • Luminescent Agents / metabolism*
  • Lung / metabolism*
  • Lung / pathology
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Transgenic
  • Promoter Regions, Genetic
  • Pulmonary Fibrosis / genetics
  • Pulmonary Fibrosis / metabolism*
  • Pulmonary Fibrosis / pathology*
  • Serine Endopeptidases / genetics*
  • Serine Endopeptidases / metabolism
  • Transcriptional Activation
  • Transforming Growth Factor beta1 / metabolism
  • Up-Regulation

Substances

  • Luminescent Agents
  • Membrane Proteins
  • Tgfb1 protein, mouse
  • Transforming Growth Factor beta1
  • Bleomycin
  • Luciferases, Firefly
  • Endopeptidases
  • Serine Endopeptidases
  • fibroblast activation protein alpha
  • Gelatinases