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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
1   Vascular Biology Program and Department of Surgery, Boston Children’s Hospital and Harvard Medical School, Boston, Massachusetts, USA
2   Paris Descartes University, Sorbonne Paris Cite, Paris, France
3   AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France
4   Inserm UMR-S1140, Faculté de Pharmacie, Paris, France
,
Marilyne Levy
5   AP-HP, Hôpital Necker-Enfants Malades, Centre de Référence Malformations Cardiaques Congénitales Complexes (M3C), Paris, France
,
Lan Huang
1   Vascular Biology Program and Department of Surgery, Boston Children’s Hospital and Harvard Medical School, Boston, Massachusetts, USA
,
Elisa Rossi
3   AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France
4   Inserm UMR-S1140, Faculté de Pharmacie, Paris, France
,
Adeline Blandinières
2   Paris Descartes University, Sorbonne Paris Cite, Paris, France
3   AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France
4   Inserm UMR-S1140, Faculté de Pharmacie, Paris, France
,
Dominique Israel-Biet
2   Paris Descartes University, Sorbonne Paris Cite, Paris, France
4   Inserm UMR-S1140, Faculté de Pharmacie, Paris, France
6   AP-HP, Hôpital Européen Georges Pompidou, Pneumology Department, Paris, France
,
Pascale Gaussem
2   Paris Descartes University, Sorbonne Paris Cite, Paris, France
3   AP-HP, Hôpital Européen Georges Pompidou, Hematology Department, Paris, France
4   Inserm UMR-S1140, Faculté de Pharmacie, Paris, France
,
Joyce Bischoff
1   Vascular Biology Program and Department of Surgery, Boston Children’s Hospital and Harvard Medical School, Boston, Massachusetts, USA
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Publikationsverlauf

Received: 01. November 2014

Accepted after major revision: 26. April 2015

Publikationsdatum:
29. November 2017 (online)

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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

 

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