title: Matrix metalloproteinase-8 promotes vascular smooth muscle cell proliferation and neointima formation.
abstract: OBJECTIVE
  We investigated the role of matrix metalloproteinase-8 (MMP8) in neointima formation and in vascular smooth muscle cell (VSMC) migration and proliferation.
  
  
  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.
authors.affiliation: From William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom (Q. Xiao, F.Z., S.Y.); Department of Cardiology, Peking University People's Hospital, Beijing, China (F.Z.); Department of Pharmacy, University of Naples Federico II, Naples, Italy (G.G., M. Maddaluno, P.M., A.I.); Cardiovascular Division, King's College London BHF Centre, London, United Kingdom (Y.H., Z.Z., Q. Xing, X.Y., M. Mayr, Q. Xu); Clemens Schöpf Institute, Technische Universität Darmstadt, Darmstadt, Germany (B.D., B.S.); Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom (P.M.).
authors.forename: Qingzhong
authors.initials: Q
authors.lastname: Xiao
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Gianluca
authors.initials: G
authors.lastname: Grassia
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Hu
authors.forename: Zhongyi
authors.initials: Z
authors.lastname: Zhang
authors.forename: Qiuru
authors.initials: Q
authors.lastname: Xing
authors.forename: Xiaoke
authors.initials: X
authors.lastname: Yin
authors.forename: Marcella
authors.initials: M
authors.lastname: Maddaluno
authors.forename: Binia
authors.initials: B
authors.lastname: Drung
authors.forename: Boris
authors.initials: B
authors.lastname: Schmidt
authors.forename: Pasquale
authors.initials: P
authors.lastname: Maffia
authors.forename: Armando
authors.initials: A
authors.lastname: Ialenti
authors.forename: Manuel
authors.initials: M
authors.lastname: Mayr
authors.forename: Qingbo
authors.initials: Q
authors.lastname: Xu
authors.forename: Shu
authors.initials: S
authors.lastname: Ye
mesh_terms: D051722:ADAM Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D015820:Cadherins / Q000378:metabolism; D020212:Carotid Artery Injuries / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D002465:Cell Movement; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000378:metabolism; D019938:Cyclin D1 / Q000378:metabolism; D004195:Disease Models, Animal; D020784:Matrix Metalloproteinase 8 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D058426:Neointima; D040901:Proteomics / Q000379:methods; D034622:RNA Interference; D013997:Time Factors; D014162:Transfection; D060449:Wnt Signaling Pathway; D051155:Wnt1 Protein / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
chemical_list: D001057:Apolipoproteins E; C495267:CTNNB1 protein, mouse; D015820:Cadherins; C530105:Ccnd1 protein, mouse; C489083:Cdh2 protein, mouse; D017077:Culture Media, Conditioned; D061965:Matrix Metalloproteinase Inhibitors; D008565:Membrane Proteins; D051155:Wnt1 Protein; C495246:Wnt1 protein, mouse; D051176:beta Catenin; D019938:Cyclin D1; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; C576328:MMP8 protein, mouse; D020784:Matrix Metalloproteinase 8; D000072197:ADAM10 Protein; C491234:Adam10 protein, mouse
keywords: matrix metalloproteinase-8; myocytes, smooth muscle; neointima