Matrix metalloproteinase-8 promotes vascular smooth muscle cell proliferation and neointima formation

Arterioscler Thromb Vasc Biol. 2014 Jan;34(1):90-8. doi: 10.1161/ATVBAHA.113.301418. Epub 2013 Oct 24.

Abstract

Objective: We investigated the role of matrix metalloproteinase-8 (MMP8) in neointima formation and in vascular smooth muscle cell (VSMC) migration and proliferation.

Approach and results: After carotid artery wire injuring, MMP8(-/-)/apoE(-/-) mice had fewer proliferating cells in neointimal lesions and smaller lesion sizes. Ex vivo assays comparing VSMCs isolated from MMP8 knockout and wild-type mice showed that MMP8 knockout decreased proliferation and migration. Proteomics analysis revealed that a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10) had lower concentrations in MMP8 knockout VSMC culture media than in MMP8 wild-type VSMC culture media. Western blot, flow cytometric, and immunocytochemical analyses showed that MMP8 knockout VSMCs contained more pro-ADAM10 but less mature ADAM10, more N-cadherin, and β-catenin in the plasma membrane but less β-catenin in the nucleus and less cyclin D1. Treatment of MMP8 wild-type VSMCs with an ADAM10 inhibitor, GI254023X, or siRNA knockdown of ADAM10 in MMP8 wild-type VSMCs inhibited proliferation and migration, increased N-cadherin and β-catenin in the plasma membrane, reduced β-catenin in the nucleus, and decreased cyclin D1 expression. Incubation of MMP8 knockout VSMCs with a recombinant ADAM10 rescued the proliferative and migratory ability of MMP8 knockout VSMCs and increased cyclin D1 expression. Furthermore, immunohistochemical analyses showed colocalization of ADAM10 with VSMCs and N-cadherin, and nuclear accumulation of β-catenin in the neointima in apoE(-/-)/MMP8(+/+) mice.

Conclusions: MMP8 enhances VSMC proliferation via an ADAM10, N-cadherin, and β-catenin-mediated pathway and plays an important role in neointima formation.

Keywords: matrix metalloproteinase-8; myocytes, smooth muscle; neointima.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / antagonists & inhibitors
  • ADAM Proteins / genetics
  • ADAM Proteins / metabolism
  • ADAM10 Protein
  • Amyloid Precursor Protein Secretases / antagonists & inhibitors
  • Amyloid Precursor Protein Secretases / genetics
  • Amyloid Precursor Protein Secretases / metabolism
  • Animals
  • Apolipoproteins E / deficiency
  • Apolipoproteins E / genetics
  • Cadherins / metabolism
  • Carotid Artery Injuries / enzymology*
  • Carotid Artery Injuries / genetics
  • Carotid Artery Injuries / pathology*
  • Cell Movement
  • Cell Proliferation* / drug effects
  • Cells, Cultured
  • Culture Media, Conditioned / metabolism
  • Cyclin D1 / metabolism
  • Disease Models, Animal
  • Matrix Metalloproteinase 8 / deficiency
  • Matrix Metalloproteinase 8 / genetics
  • Matrix Metalloproteinase 8 / metabolism*
  • Matrix Metalloproteinase Inhibitors / pharmacology
  • Membrane Proteins / antagonists & inhibitors
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Muscle, Smooth, Vascular / drug effects
  • Muscle, Smooth, Vascular / enzymology*
  • Muscle, Smooth, Vascular / pathology
  • Myocytes, Smooth Muscle / drug effects
  • Myocytes, Smooth Muscle / enzymology*
  • Myocytes, Smooth Muscle / pathology
  • Neointima*
  • Proteomics / methods
  • RNA Interference
  • Time Factors
  • Transfection
  • Wnt Signaling Pathway
  • Wnt1 Protein / metabolism
  • beta Catenin / metabolism

Substances

  • Apolipoproteins E
  • CTNNB1 protein, mouse
  • Cadherins
  • Ccnd1 protein, mouse
  • Cdh2 protein, mouse
  • Culture Media, Conditioned
  • Matrix Metalloproteinase Inhibitors
  • Membrane Proteins
  • Wnt1 Protein
  • Wnt1 protein, mouse
  • beta Catenin
  • Cyclin D1
  • Amyloid Precursor Protein Secretases
  • ADAM Proteins
  • MMP8 protein, mouse
  • Matrix Metalloproteinase 8
  • ADAM10 Protein
  • Adam10 protein, mouse