Abnormal activation of calpain and protein kinase Cα promotes a constitutive release of matrix metalloproteinase 9 in peripheral blood mononuclear cells from cystic fibrosis patients

Arch Biochem Biophys. 2016 Aug 15:604:103-12. doi: 10.1016/j.abb.2016.06.015. Epub 2016 Jun 25.

Abstract

Matrix metalloproteinase 9 (MMP9) is physiologically involved in remodeling the extracellular matrix components but its abnormal release has been observed in several human pathologies. We here report that peripheral blood mononuclear cells (PBMCs), isolated from cystic fibrosis (CF) patients homozygous for F508del-cystic fibrosis transmembrane conductance regulator (CFTR), express constitutively and release at high rate MMP9 due to the alteration in their intracellular Ca(2+) homeostasis. This spontaneous and sustained MMP9 secretion may contribute to the accumulation of this protease in fluids of CF patients. Conversely, in PBMCs isolated from healthy donors, expression and secretion of MMP9 are undetectable but can be evoked, after 12 h of culture, by paracrine stimulation which also promotes an increase in [Ca(2+)]i. We also demonstrate that in both CF and control PBMCs the Ca(2+)-dependent MMP9 secretion is mediated by the concomitant activation of calpain and protein kinase Cα (PKCα), and that MMP9 expression involves extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) phosphorylation. Our results are supported by the fact that either the inhibition of Ca(2+) entry or chelation of [Ca(2+)]i as well as the inhibition of single components of the signaling pathway or the restoration of CFTR activity all promote the reduction of MMP9 secretion.

Keywords: Calcium; Calpain; Cystic fibrosis; MMP9; PBMCs; PKC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Calcium / metabolism
  • Calpain / blood*
  • Cystic Fibrosis / blood*
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics*
  • Enzyme Activation
  • Epithelial Cells / cytology
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Gene Deletion
  • Gene Expression Regulation, Enzymologic
  • Homeostasis
  • Homozygote
  • Humans
  • Leukocytes, Mononuclear / metabolism*
  • Matrix Metalloproteinase 9 / blood*
  • Middle Aged
  • Phosphorylation
  • Predictive Value of Tests
  • Protein Kinase C-alpha / blood*
  • Young Adult

Substances

  • cystic fibrosis transmembrane conductance regulator delta F508
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • PRKCA protein, human
  • Protein Kinase C-alpha
  • Extracellular Signal-Regulated MAP Kinases
  • Calpain
  • MMP9 protein, human
  • Matrix Metalloproteinase 9
  • Calcium