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Heparin affin regulatory peptide binds to vascular endothelial growth factor (VEGF) and inhibits VEGF-induced angiogenesis

Oncogene volume 23, pages 1745–1753 (2004)Cite this article

Abstract

Heparin affin regulatory peptide (HARP) is an heparin-binding molecule involved in the regulation of cell proliferation and differentiation. Here, we report that HARP inhibited the biological activity induced by the 165-amino-acid form of vascular endothelial growth factor (VEGF165) on human umbilical vein endothelial cells. Endothelial-cell proliferation induced by VEGF165 showed about 50% inhibition in the presence of HARP in a concentration of 3 nM. In similar range of concentrations, HARP blocked tube formation induced by VEGF165 in three-dimensional angiogenesis assay. In vivo studies showed that HARP inhibited the VEGF165-induced Matrigel™ infiltration of endothelial cells. We then investigated the mechanisms of this inhibition and shown that HARP inhibited the binding of 125I-VEGF165 to the VEGF receptors of endothelial cells. Additional studies using VEGF soluble receptors indicated that binding of 125I-VEGF165 to kinase insert domain-containing receptor and neuropilin receptor was inhibited by HARP, but conversely the binding of 125I-VEGF165 to fms-like tyrosine kinase I receptor was unaffected. A competitive affinity-binding assay demonstrated that HARP interacted directly with VEGF165 with a dissociation coefficient of 1.38 nM. Binding assay using deletion mutants of HARP revealed that the thrombospondin type-1 repeats domains were involved in this interaction. These data demonstrate for the first time that the angiogenic factor HARP can also negatively regulates the angiogenic activity of VEGF165.

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Abbreviations

HARP:

heparin affin regulatory peptide

VEGF165:

165-amino-acid form of vascular endothelial growth factor

HUVECs:

human umbilical vein endothelial cells

TSR:

thrombospondin repeat

PBS:

phosphate buffer saline

DSS:

disuccinimidyl suberate

TSP-1:

thrombospondin 1

flt-1:

fms-like tyrosine kinase-1

KDR:

kinase insert domain containing receptor, NP-1 neuropilin-1

GST:

glutathione S-transferase

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Acknowledgements

This work was supported by grants from four non-profit sources, namely, the Association pour la Recherche sur le Cancer (No. 4248), the Ministère de l'Education Nationale (DRED), the CNRS, and the Research Committee of the University of Patras. We thank Dr D Caruelle for helpful discussions and critically reading this manuscript. MH received a grant from a non-profit organization, the Association pour la Recherche sur la Polyarthrite.

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  1. Contributed equally to this work

Authors and Affiliations

  1. Laboratoire de Recherche sur la Croissance Cellulaire, la Réparation et la Régénération Tissulaires (CRRET), FRE CNRS 2412, Université Paris XII-Val de Marne, Avenue du Général de Gaulle, Créteil, 94010, France

    Mélanie Héroult, Isabelle Bernard-Pierrot, Jean Delbé, Yamina Hamma-Kourbali, Denis Barritault & José Courty

  2. Division of Genetics, Department of Biology, Cell and Developmental Biology, University of Patras, Patras, GR-26504, Greece

    Panagiotis Katsoris

  3. Department of Pharmacy, Laboratory of Molecular Pharmacology, University of Patras, Patras, 26504, GR, Greece

    Evangelia Papadimitriou

  4. Laboratoire de Biologie Moléculaire Eucaryote, CNRS UPR 9006, 118 Route de Narbonne, Toulouse, 31062, France

    Jean Plouet

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  1. Mélanie Héroult
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Correspondence to José Courty.

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Héroult, M., Bernard-Pierrot, I., Delbé, J. et al. Heparin affin regulatory peptide binds to vascular endothelial growth factor (VEGF) and inhibits VEGF-induced angiogenesis. Oncogene 23, 1745–1753 (2004). https://doi.org/10.1038/sj.onc.1206879

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