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Mechanistic study of the proangiogenic effect of osteoprotegerin

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Abstract

Osteoprotegerin (OPG), a soluble tumour necrosis factor receptor superfamily member, inhibits RANKL-mediated osteoclastogenesis. We have previously reported that OPG enhances the proangiogenic properties of endothelial colony-forming cells (ECFCs) in vitro, and promotes vasculogenesis in vivo. Here we investigated how OPG promotes neovascularisation. Proteomic experiments showed that OPG pretreatment affected ECFCs protein expression in two ways, 23 spots being down-regulated and 6 upregulated. These spots corresponded to proteins involved in cell motility, adhesion, signal transduction and apoptosis. In keeping with these proteomic results, we found that OPG induced ECFCs adhesion to activated endothelium in shear stress conditions, promoting intermediate but not focal adhesion to fibronectin and collagen. Treatment with OPG induced a reorganization of the ECFCs cytoskeleton, with the emergence of cell protrusions characteristic of a migratory phenotype. These effects correlated with decreased FAK phosphorylation and enhanced integrin αVβ3 expression. OPG drastically reduced caspase-3/7 activities and maintained ECFCs viability after 48 h of treatment. All these effects were significantly attenuated by ECFCs incubation with the CXCR4 antagonist AMD-3100, and by prior heparan sulphate proteoglycan disruption. The proangiogenic properties of OPG appeared to be mediated by the proteoglycan syndecan-1, although OPG 1-194 lacking its heparin-binding domain still had pro-vasculogenic effects in vitro and in vivo. These results suggest that OPG may interact with ECFCs by binding to HSPGs/syndecan-1, thereby induce an anti-adhesive effect and promoting ECFCs migration through a SDF-1/CXCR4 dependent pathway.

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Acknowledgments

We thank C. Martin and the technicians from the IMTCE animal facilities (Paris Descartes University). We are also indebted to the nursing services of Hôpital des Diaconnesses (Paris) and Hôpital des Instructions et des Armées de Begin (Saint Mandé) for providing umbilical cord blood samples. This work was supported in part by research grants from Institut Français de la Recherche sur la Mer (08/5210807). Z. Benslimane-Ahmim received grants from Groupe d’Etude et de Recherches sur l’Hémostase (GEHT). CNRS pays the salary of C. Boisson-Vidal.

Conflict of interest

  The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Université Paris Descartes, Sorbonne Paris Cité, Paris, France

    Zahia Benslimane-Ahmim, Anna Lokajczyk, Françoise Grelac, Isabelle Galy-Fauroux, Anne-Marie Fischer & Catherine Boisson-Vidal

  2. INSERM, UMR-S765, Paris, France

    Zahia Benslimane-Ahmim, Anna Lokajczyk, Françoise Grelac, Isabelle Galy-Fauroux, Anne-Marie Fischer & Catherine Boisson-Vidal

  3. LBPS, UFR SMBH Léonard de Vinci, CNRS CSPBAT, Bobigny, France

    Florence Poirier & Didier Lutomski

  4. Université Paris 13, Sorbonne Paris Cité, 74 Rue Marcel Cachin, Bobigny, France

    Florence Poirier, Amel Mohamedi & Didier Lutomski

  5. Université Paris-Sud–IFR141 IPSIT–Trans-Prot-5, rue Jean-Baptiste Clément, 92296, Chatenay-Malabry, France

    Claudine Delomenie

  6. Haematology Department, AP-HP, Hôpital European Georges Pompidou, Paris, France

    Anne-Marie Fischer

  7. Equipe LIGUE 2012, INSERM, UMR-S 957, Nantes, France

    Dominique Heymann

  8. Université de Nantes, Nantes Atlantique University, Nantes, France

    Dominique Heymann

  9. CHU Hôtel Dieu, Nantes, France

    Dominique Heymann

  10. Faculté de Pharmacie 4 av. de l’Observatoire, INSERM, U765, 75006, Paris, France

    Zahia Benslimane-Ahmim

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  1. Zahia Benslimane-Ahmim
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  2. Florence Poirier
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Correspondence to Zahia Benslimane-Ahmim.

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Zahia Benslimane-Ahmim and Florence Poirier contributed equally to this study.

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Benslimane-Ahmim, Z., Poirier, F., Delomenie, C. et al. Mechanistic study of the proangiogenic effect of osteoprotegerin. Angiogenesis 16, 575–593 (2013). https://doi.org/10.1007/s10456-013-9337-x

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