title: High urokinase expression contributes to the angiogenic properties of endothelial cells derived from circulating progenitors.
abstract: Endothelial progenitor cells (EPC) display a unique ability to repair vascular injury and promote neovascularization although the underlying molecular mechanisms remain poorly understood. Urokinase-type plasminogen activator (uPA) and its receptor (uPAR) play a critical role in cell migration and angiogenesis by facilitating proteolysis of extracellular matrix. The aim of the present study was to characterize the uPA/uPAR-dependent proteolytic potential of EPC outgrown from human umbilical cord blood and to analyze its contribution to their angiogenic properties in vitro. Cells derived from EPC (EPDC), presenting typical features of late outgrowth endothelial cells, were compared to mature endothelial cells, represented by human umbilical vein endothelial cells (HUVEC). Using quantitative flow cytometry, enzyme-linked immunosorbent assays and zymography, we demonstrated that EPDC displayed higher levels of uPA and uPAR. In conditioned culture media, uPA-dependent proteolytic activity was also found to be significantly increased in EPDC. This activity was paralleled by a higher secretion of pro-metalloproteinase-2 (pro-MMP-2). Inhibition of EPDC-associated uPA by monoclonal antibodies that block either uPA activity or receptor binding, significantly reduced proliferation, migration and capillary like tube formation. Moreover, tumor necrosis factor-alpha and vascular endothelial growth factor, known to be locally secreted in ischemic areas, further increased the proteolytic potential of EPDC by up-regulating uPA and uPAR expression respectively. The EPDC response to these factors was found to be more pronounced than that of HUVEC. In conclusion, these findings indicated that EPDC are characterized by high intrinsic uPA/uPAR-dependent proteolytic potential that could contribute to their invasive and angiogenic behaviour.
authors.affiliation: INSERM U608, Laboratoire d'Hématologie et d'Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille Cedex 5, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Basire
authors.forename: Florence
authors.initials: F
authors.lastname: Sabatier
authors.forename: Sophie
authors.initials: S
authors.lastname: Ravet
authors.forename: Edouard
authors.initials: E
authors.lastname: Lamy
authors.forename: Agnès
authors.initials: A
authors.lastname: Mialhe
authors.forename: Gwladys
authors.initials: G
authors.lastname: Zabouo
authors.forename: Pascale
authors.initials: P
authors.lastname: Paul
authors.forename: Victor
authors.initials: V
authors.lastname: Gurewich
authors.forename: José
authors.initials: J
authors.lastname: Sampol
authors.forename: Françoise
authors.initials: F
authors.lastname: Dignat-George
mesh_terms: D002465:Cell Movement; D049109:Cell Proliferation; D004730:Endothelium, Vascular / Q000473:pathology*; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D007511:Ischemia / Q000473:pathology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D018919:Neovascularization, Physiologic; D011956:Receptors, Cell Surface / Q000096:biosynthesis*; D055293:Receptors, Urokinase Plasminogen Activator; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis* / Q000378:metabolism*; D014945:Wound Healing
chemical_list: C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D020778:Matrix Metalloproteinase 2