High urokinase expression contributes to the angiogenic properties of endothelial cells derived from circulating progenitors

Agnès Basire; Florence Sabatier; Sophie Ravet; Edouard Lamy; Agnès Mialhe; Gwladys Zabouo; Pascale Paul; Victor Gurewich; José Sampol; Françoise Dignat-George

doi:10.1160/TH05-07-0511

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2006; 95(04): 678-688
DOI: 10.1160/TH05-07-0511

Endothelium and Vascular Development

Schattauer GmbH

High urokinase expression contributes to the angiogenic properties of endothelial cells derived from circulating progenitors

Agnès Basire

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

²Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France

,

Florence Sabatier

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

²Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France

,

Sophie Ravet

²Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France

,

Edouard Lamy

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

,

Agnès Mialhe

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

,

Gwladys Zabouo

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

,

Pascale Paul

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

²Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France

,

Victor Gurewich

³Vascular Research Laboratory, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

,

José Sampol

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

,

Françoise Dignat-George

¹INSERM U608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France

²Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France

› Author Affiliations

Abstract

Summary

Endothelial progenitor cells (EPC) displaya 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-dependant proteolytic activity was also found to be significantly increased in EPDC.This activity was paralleled bya 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 factoralpha 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.

Keywords

Endothelial progenitor - urokinase - angiogenesis - proteolysis

Full Text

References

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