Treprostinil indirectly regulates endothelial colony forming cell angiogenic properties by increasing VEGF-A produced by mesenchymal stem cells

David M. Smadja; Marilyne Levy; Lan Huang; Elisa Rossi; Adeline Blandinières; Dominique Israel-Biet; Pascale Gaussem; Joyce Bischoff

doi:10.1160/TH14-11-0907

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2015; 114(04): 735-747
DOI: 10.1160/TH14-11-0907

Endothelium and Angiogenesis

Schattauer GmbH

Treprostinil indirectly regulates endothelial colony forming cell angiogenic properties by increasing VEGF-A produced by mesenchymal stem cells

David M. Smadja

¹Vascular Biology Program and Department of Surgery, Boston Children’s Hospital and Harvard Medical School, Boston, Massachusetts, USA

²Paris Descartes University, Sorbonne Paris Cite, Paris, France

³AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France

⁴Inserm UMR-S1140, Faculté de Pharmacie, Paris, France

,

Marilyne Levy

⁵AP-HP, Hôpital Necker-Enfants Malades, Centre de Référence Malformations Cardiaques Congénitales Complexes (M3C), Paris, France

,

Lan Huang

¹Vascular Biology Program and Department of Surgery, Boston Children’s Hospital and Harvard Medical School, Boston, Massachusetts, USA

,

Elisa Rossi

³AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France

⁴Inserm UMR-S1140, Faculté de Pharmacie, Paris, France

,

Adeline Blandinières

²Paris Descartes University, Sorbonne Paris Cite, Paris, France

³AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France

⁴Inserm UMR-S1140, Faculté de Pharmacie, Paris, France

,

Dominique Israel-Biet

²Paris Descartes University, Sorbonne Paris Cite, Paris, France

⁴Inserm UMR-S1140, Faculté de Pharmacie, Paris, France

⁶AP-HP, Hôpital Européen Georges Pompidou, Pneumology Department, Paris, France

,

Pascale Gaussem

²Paris Descartes University, Sorbonne Paris Cite, Paris, France

³AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France

⁴Inserm UMR-S1140, Faculté de Pharmacie, Paris, France

,

Joyce Bischoff

¹Vascular Biology Program and Department of Surgery, Boston Children’s Hospital and Harvard Medical School, Boston, Massachusetts, USA

› Author Affiliations

Abstract

Summary

Pulmonary vasodilators and prostacyclin therapy in particular, have markedly improved the outcome of patients with pulmonary hypertension (PH). Endothelial dysfunction is a key feature of PH, and we previously reported that treprostinil therapy increases number and proliferative potential of endothelial colony forming cells (ECFC) isolated from PH patients’ blood. In the present study, the objective was to determine how treprostinil contributes to the proangiogenic functions of ECFC. We examined the effect of treprostinil on ECFC obtained from cord blood in terms of colony numbers, proliferative and clonogenic properties in vitro, as well as in vivo vasculogenic properties. Surprisingly, treprostinil inhibited viability of cultured ECFC but did not modify their clonogenic properties or the endothelial differentiation potential from cord blood stem cells. Treprostinil treatment significantly increased the vessel-forming ability of ECFC combined with mesenchymal stem cells (MSC) in Matrigel implanted in nude mice. In vitro, ECFC proliferation was stimulated by conditioned media from treprostinil-pretreated MSC, and this effect was inhibited either by the use of VEGF-A blocking antibodies or siRNA VEGF-A in MSC. Silencing VEGF-A gene in MSC also blocked the pro-angiogenic effect of treprostinil in vivo. In conclusion, increased VEGF-A produced by MSC can account for the increased vessel formation observed during treprostinil treatment. The clinical relevance of these data was confirmed by the high level of VEGF-A detected in plasma from patients with paediatric PH who had been treated with treprostinil. Moreover, our results suggest that VEGF-A level in patients could be a surrogate biomarker of treprostinil efficacy.

Keywords

Stem cells - vascular remodelling - vasculogenesis - endothelial progenitor cells - prostacyclin analogues

Full Text

References

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