Fibronectin fibrils regulate TGF-β1-induced Epithelial-Mesenchymal Transition

Matrix Biol. 2017 Jul;60-61:157-175. doi: 10.1016/j.matbio.2017.01.001. Epub 2017 Jan 19.

Abstract

Epithelial-Mesenchymal Transition (EMT) is a dynamic process through which epithelial cells transdifferentiate from an epithelial phenotype into a mesenchymal phenotype. Previous studies have demonstrated that both mechanical signaling and soluble growth factor signaling facilitate this process. One possible point of integration for mechanical and growth factor signaling is the extracellular matrix. Here we investigate the role of the extracellular matrix (ECM) protein fibronectin (FN) in this process. We demonstrate that inhibition of FN fibrillogenesis blocks activation of the Transforming Growth Factor-Beta (TGF-β) signaling pathway via Smad2 signaling, decreases cell migration and ultimately leads to inhibition of EMT. Results show that soluble FN, FN fibrils, or increased contractile forces are insufficient to independently induce EMT. We further demonstrate that inhibition of latent TGF-β1 binding to FN fibrils via either a monoclonal blocking antibody against the growth factor binding domain of FN or through use of a FN deletion mutant that lacks the growth factor binding domains of FN blocks EMT progression, indicating a novel role for FN in EMT in which the assembly of FN fibrils serves to localize TGF-β1 signaling to drive EMT.

Keywords: Epithelial-Mesenchymal Transition; Extracellular matrix; Fibronectin; TGF-β1.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / pharmacology
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / pharmacology
  • Biomechanical Phenomena
  • Cell Line, Tumor
  • Cell Movement
  • Cytokines / antagonists & inhibitors
  • Cytokines / chemistry
  • Cytokines / genetics
  • Cytokines / metabolism*
  • Dogs
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Epithelial-Mesenchymal Transition / drug effects*
  • Extracellular Matrix / chemistry
  • Extracellular Matrix / drug effects*
  • Extracellular Matrix / metabolism
  • Fibronectins
  • Gene Expression Regulation
  • Humans
  • Madin Darby Canine Kidney Cells
  • Mutation
  • Protein Binding / drug effects
  • Signal Transduction
  • Smad2 Protein / genetics
  • Smad2 Protein / metabolism
  • Streptococcus pyogenes / chemistry
  • Streptococcus pyogenes / metabolism
  • Transforming Growth Factor beta1 / metabolism
  • Transforming Growth Factor beta1 / pharmacology*

Substances

  • Antibodies, Monoclonal
  • Bacterial Proteins
  • Cytokines
  • FN1 protein, human
  • Fibronectins
  • SMAD2 protein, human
  • Smad2 Protein
  • TGFB1 protein, human
  • Transforming Growth Factor beta1