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Thromb Haemost 2005; 94(06): 1270-1279
DOI: 10.1160/TH05-07-0499
Endothelium and Vascular Development
Schattauer GmbH

Presence of endothelial progenitor cells, distinct from mature endothelial cells, within human CD146+ blood cells

Bruno Delorme
1   INSERM Unité 608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France
,
Agnès Basire
1   INSERM Unité 608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France
,
Carla Gentile
3   INSERM Unité 602, Groupe Hospitalier Paul Brousse, Villejuif, France
,
Florence Sabatier
1   INSERM Unité 608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France
2   Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France
,
Frédéric Monsonis
4   Beckman Coulter Immunotech Company, Marseille, France
,
Christophe Desouches
5   Service de Chirurgie Plastique, Réparatrice et Esthétique, Centre Hospitalier Universitaire La Conception, Marseille, France
,
Marcel Blot-Chabaud
1   INSERM Unité 608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France
,
Georges Uzan
3   INSERM Unité 602, Groupe Hospitalier Paul Brousse, Villejuif, France
,
José Sampol
1   INSERM Unité 608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France
2   Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France
,
Françoise Dignat-George
1   INSERM Unité 608, Laboratoire d’Hématologie et d’Immunologie, UFR de Pharmacie, Université de la Méditerranée, Marseille, France
2   Laboratoire d’Hématologie, Centre Hospitalier Universitaire La Conception, Marseille, France
› Author AffiliationsFinancial support: Supported by the Institut National de la Santé et de la Recherche Médicale (network Biotherapie No 4CS01H) and in part by research funding from Sanofi-Synthelabo.
Further Information

Publication History

Received 18 July 2005

Accepted after revision 13 October 2005

Publication Date:
07 December 2017 (online)

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Summary

CD146 is an adhesion molecule present on endothelial cells throughout the vascular tree. CD146 is also expressed by circulating endothelial cells (CECs) widely considered to be mature endothelial cells detached from injured vessels. The discovery of circulating endothelial progenitor cells (EPCs) originating from bone marrow prompted us to investigate whether CD146 circulating cells could also contains EPCs. We tested this hypothesis using an approach combining elimination of CECs by an adhesion step, followed by immunomagnetic sorting of remaining CD146+ cells from the non adherent fraction of cord blood mononuclear cells. When cultured under endothelial-promoting conditions, these cells differentiated as late outgrowth endothelial colonies: they grew as a cobblestone monolayer, were uniformly positive for endothelial markers and did not express leukocyte antigens. They highly proliferated and were expanded in long-term culture without alterations of their phenotypic and functional properties (DiI-ac-LDL uptake, wound repair, capillary-like network formation, and TNFα response). Moreover, these cells colonized a Matrigel plug in immunodeficient mice (NOD/SCID). Finally, using 4-color flow cytometry analysis of purified CD34+ cells, we clearly discriminated, CD146+ EPCs (CD146+ CD34+ CD45+ CD133+ or CD117+), and CD146+ CECs (CD146+ CD34+, CD45− CD133− or CD117−), both in cord and adult peripheral blood. The relative proportions of the two CD146+ subsets varied in patients with myocardial infarction as compared to healthy subjects. Our study establishes that, beside CECs, CD146+ circulating cells contain a subpopulation of EPCs with potential use in proangiogenic therapy. In addition, the dual measurement of CD146+ CECs and CD146+ EPCs offers a promising tool for monitoring vascular injury/regeneration processes in clinical situations.

Keywords

CD146 - angiogenesis - myocardial infarction - circulating endothelial cells - progenitor endothelial cells
 

  • References

  • 1 Bardin N. et al S-Endo 1, a pan-endothelial monoclonal antibody recognizing a novel human endothelial antigen. Tissue Antigens 1996; 48: 531-9.
    CrossrefPubMedGoogle Scholar
  • 2 Bardin N. et al Identification of the S-Endo 1 endothelial-associated antigen. Biochem Biophys Res Commun 1996; 218: 210-6.
    CrossrefPubMedGoogle Scholar
  • 3 Bardin N. et al Identification of CD146 as a component of the endothelial junction involved in the control of cell-cell cohesion. Blood 2001; 98: 3677-84.
    CrossrefPubMedGoogle Scholar
  • 4 Anfosso F. et al Activation of human endothelial cells via S-endo-1 antigen (CD146) stimulates the tyrosine phosphorylation of focal adhesion kinase p125(FAK). J Biol Chem 1998; 273: 26852-6.
    CrossrefPubMedGoogle Scholar
  • 5 Anfosso F. et al Outside-in signaling pathway linked to CD146 engagement in human endothelial cells. J Biol Chem 2001; 276: 1564-9.
    CrossrefPubMedGoogle Scholar
  • 6 Blann AD. et al Circulating endothelial cells : biomarker of vascular disease. Thromb Haemost 2005; 93: 228-35.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 George F. et al Demonstration of Rickettsia conorii-induced endothelial injury in vivo by measuring circulating endothelial cells, thrombomodulin, and von Willebrand factor in patients with Mediterranean spotted fever. Blood 1993; 82: 2109-16.
    PubMedGoogle Scholar
  • 8 Quilici J. et al Circulating endothelial cell count as a diagnostic marker for non-ST-elevation acute coronary syndromes. Circulation 2004; 110: 1586-91.
    CrossrefPubMedGoogle Scholar
  • 9 Woywodt A. et al Circulating endothelial cells as markers for ANCA-associated small-vessel vasculitis. Lancet 2003; 361: 206-10.
    CrossrefPubMedGoogle Scholar
  • 10 Solovey A. et al Circulating activated endothelial cells in sickle cell anemia. N Engl J Med 1997; 337: 1584-90.
    CrossrefPubMedGoogle Scholar
  • 11 Popa ER. et al Donor-derived circulating endothelial cells after kidney transplantation. Transplantation 2002; 74: 1320-7.
    CrossrefPubMedGoogle Scholar
  • 12 Dignat-George F, Sampol J. Circulating endothelial cells in vascular disorders: new insights into an old concept. Eur J Haematol 2000; 65: 215-20.
    CrossrefPubMedGoogle Scholar
  • 13 Rajagopalan S. et al Endothelial cell apoptosis in systemic lupus erythematosus: a common pathway for abnormal vascular function and thrombosis propensity. Blood 2004; 103: 3677-83.
    CrossrefPubMedGoogle Scholar
  • 14 Clancy R. et al Circulating activated endothelial cells in systemic lupus erythematosus: further evidence for diffuse vasculopathy. Arthritis Rheum 2001; 44: 1203-8.
    CrossrefPubMedGoogle Scholar
  • 15 Beerepoot LV. et al Increased levels of viable circulating endothelial cells are an indicator of progressive disease in cancer patients. Ann Oncol 2004; 15: 139-45.
    CrossrefPubMedGoogle Scholar
  • 16 Rafii S, Lyden D. Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. Nat Med 2003; 9: 702-12.
    CrossrefPubMedGoogle Scholar
  • 17 Asahara T. et al Isolation of putative progenitor endothelial cells for angiogenesis. Science 1997; 275: 964-7.
    CrossrefPubMedGoogle Scholar
  • 18 Lin Y. et al Origins of circulating endothelial cells and endothelial outgrowth from blood. J Clin Invest 2000; 105: 71-7.
    CrossrefPubMedGoogle Scholar
  • 19 Gulati R. et al Diverse origin and function of cells with endothelial phenotype obtained from adult human blood. Circ Res 2003; 93: 1023-5.
    CrossrefPubMedGoogle Scholar
  • 20 Hur J, Yoon CH, Kim HS. et al Characterization of two types of endothelial progenitor cells and their different contributions to neovasculogenesis. Arterioscler Thromb Vasc Biol 2004; 24: 288-93.
    CrossrefPubMedGoogle Scholar
  • 21 Hill JM, Zalos G, Halcox JP. et al Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. N Engl J Med 2003; 348: 593-600.
    CrossrefPubMedGoogle Scholar
  • 22 Choi JH. et al Decreased number and impaired angiogenic function of endothelial progenitor cells in patients with chronic renal failure. Arterioscler Thromb Vasc Biol 2004; 24: 1246-52.
    CrossrefPubMedGoogle Scholar
  • 23 Vasa M. et al Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease. Circ Res 2001; 89: E1-7.
    CrossrefPubMedGoogle Scholar
  • 24 Woywodt A. et al Circulating endothelial cells as a marker of endothelial damage in allogeneic hematopoietic stem cell transplantation. Blood 2004; 103: 3603-5.
    CrossrefPubMedGoogle Scholar
  • 25 Burger PE. et al Fibroblast growth factor receptor-1 is expressed by endothelial progenitor cells. Blood 2002; 100: 3527-35.
    CrossrefPubMedGoogle Scholar
  • 26 Rehman J. et al Peripheral blood “endothelial progenitor cells” are derived from monocyte/macrophages and secrete angiogenic growth factors. Circulation 2003; 107: 1164-9.
    CrossrefPubMedGoogle Scholar
  • 27 Schmeisser A. et al Monocytes coexpress endothelial and macrophagocytic lineage markers and form cord-like structures in Matrigel under angiogenic conditions. Cardiovasc Res 2001; 49: 671-80.
    CrossrefPubMedGoogle Scholar
  • 28 Miraglia S. et al A novel five-transmembrane hematopoietic stem cell antigen: isolation, characterization, and molecular cloning. Blood 1997; 90: 5013-21.
    Google Scholar
  • 29 Kalka C. et al Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci U S A 2000; 97: 3422-7.
    CrossrefPubMedGoogle Scholar
  • 30 Kocher AA. et al Neovascularization of ischemic myocardium by human bone-marrow-derived angioblasts prevents cardiomyocyte apoptosis, reduces remodeling and improves cardiac function. Nat Med 2001; 7: 430-6.
    CrossrefPubMedGoogle Scholar
  • 31 Tateishi-Yuyama E. et al Therapeutic angiogenesis for patients with limb ischaemia by autologous transplantation of bone-marrow cells: a pilot study and a randomised controlled trial. Lancet 2002; 360: 427-35.
    CrossrefPubMedGoogle Scholar
  • 32 Assmus B. et al Transplantation of progenitor cells and regeneration enhancement in acute myocardial infarction. Circulation 2002; 106: 3009-17.
    CrossrefPubMedGoogle Scholar
  • 33 Kudo FA. et al Autologous transplantation of peripheral blood endothelial progenitor cells (CD34+) for therapeutic angiogenesis in patients with critical limb ischemia. Int Angiol 2003; 22: 344-8.
    PubMedGoogle Scholar
  • 34 Peichev M, Naiyer AJ, Pereira D. et al Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors. Blood 2000; 95: 952-8.
    PubMedGoogle Scholar
  • 35 Gehling UM. et al In vitro differentiation of endothelial cells from AC133-positive progenitor cells. Blood 2000; 95: 3106-12.
    PubMedGoogle Scholar
  • 36 Bompais H. et al Human endothelial cells derived from circulating progenitors display specific functional properties compared with mature vessel wall endothelial cells. Blood 2004; 103: 2577-84.
    CrossrefPubMedGoogle Scholar
  • 37 Pelosi E. et al Identification of the hemangioblast in postnatal life. Blood 2002; 100: 3203-8.
    CrossrefPubMedGoogle Scholar
  • 38 Schatteman GC, Awad O. Hemangioblasts, angioblasts, and adult endothelial cell progenitors. Anat Rec 2004; 276A: 13-21.
    CrossrefPubMedGoogle Scholar
  • 39 Grant MB. et al Adult hematopoietic stem cells provide functional hemangioblast activity during retinal neovascularization. Nat Med 2002; 8: 607-12.
    CrossrefPubMedGoogle Scholar
  • 40 Filshie RJ. et al MUC18, a member of the immunoglobulin superfamily, is expressed on bone marrow fibroblasts and a subset of hematological malignancies. Leukemia 1998; 12: 414-21.
    CrossrefPubMedGoogle Scholar
  • 41 Ingram DA. et al Identification of a novel hierarchy of endothelial progenitor cells using peripheral and umbilical cord blood. Blood 2004; 104: 2752-60.
    CrossrefPubMedGoogle Scholar
  • 42 Alais S. et al HEMCAM/CD146 downregulates cell surface expression of β1 integrins. J Cell Sci 2001; 114: 1847-59.
    PubMedGoogle Scholar
  • 43 Solovey AN. et al Identification and functional assessment of endothelial P1H12. J Lab Clin Med 2001; 138: 322-31.
    CrossrefPubMedGoogle Scholar
  • 44 Yan X. et al A novel anti-CD146 monoclonal antibody, AA98, inhibits angiogenesis and tumor growth. Blood 2003; 102: 184-91.
    CrossrefPubMedGoogle Scholar
  • 45 Shintani S. et al Mobilization of endothelial progenitor cells in patients with acute myocardial infarction. Circulation 2001; 103: 2776-9.
    CrossrefPubMedGoogle Scholar
  • 46 Tepper OM. et al Adult vasculogenesis occurs through the in situ recruitment, proliferation and tubulization of circulating bone marrow-derived cells. Blood 2004; 105: 1068-77.
    CrossrefPubMedGoogle Scholar