Clinical & Experimental Metastasis

, Volume 25, Issue 1, pp 53–64

A novel αvβ3-blocking disintegrin containing the RGD motive, DisBa-01, inhibits bFGF-induced angiogenesis and melanoma metastasis

Authors

  • Oscar H. P. Ramos
    • Dep. Ciências FisiológicasUniversidade Federal de São Carlos
  • Alexandre Kauskot
    • INSERM, U 553
    • Université Paris 7-Denis Diderot, Faculté de Médecine, IFR 105 -Saint Louis-Institut Universitaire d’Hématologie
  • Márcia R. Cominetti
    • Dep. Ciências FisiológicasUniversidade Federal de São Carlos
  • Iga Bechyne
    • INSERM, U 553
    • Université Paris 7-Denis Diderot, Faculté de Médecine, IFR 105 -Saint Louis-Institut Universitaire d’Hématologie
  • Carmen L. Salla Pontes
    • Dep. Ciências FisiológicasUniversidade Federal de São Carlos
  • Fabrice Chareyre
    • Université Paris 7-Denis Diderot, Faculté de Médecine, IFR 105 -Saint Louis-Institut Universitaire d’Hématologie
    • INSERM, U 674
  • Jan Manent
    • Université Paris 7-Denis Diderot, Faculté de Médecine, IFR 105 -Saint Louis-Institut Universitaire d’Hématologie
    • INSERM, U 674
  • Roger Vassy
    • Laboratoire de Pharmacologie Clinique et ExperimentaleUniversité Paris 13, CNRS UMR 7033
  • Marco Giovannini
    • Université Paris 7-Denis Diderot, Faculté de Médecine, IFR 105 -Saint Louis-Institut Universitaire d’Hématologie
    • INSERM, U 674
  • Chantal Legrand
    • INSERM, U 553
    • Université Paris 7-Denis Diderot, Faculté de Médecine, IFR 105 -Saint Louis-Institut Universitaire d’Hématologie
  • Heloisa S. Selistre-de-Araujo
    • Dep. Ciências FisiológicasUniversidade Federal de São Carlos
  • Michel Crépin
    • INSERM, U 553
    • Laboratoire de Pharmacologie Clinique et ExperimentaleUniversité Paris 13, CNRS UMR 7033
    • INSERM, U 553
    • Université Paris 7-Denis Diderot, Faculté de Médecine, IFR 105 -Saint Louis-Institut Universitaire d’Hématologie
    • Laboratoire d’Immunologie, Centre de Recherche du Centre Hospitalier de l’Université de Montréal-Saint-Luc, INSERM U743
Research Paper

DOI: 10.1007/s10585-007-9101-y

Cite this article as:
Ramos, O.H.P., Kauskot, A., Cominetti, M.R. et al. Clin Exp Metastasis (2008) 25: 53. doi:10.1007/s10585-007-9101-y

Abstract

The integrin αvβ3 is involved in multiple aspects of malignant cancer, including tumor angiogenesis and metastasis, which makes the receptor a key target for the development of anti-cancer therapies. We report here on the production, the characterization and the in vivo anti-angiogenic and anti-metastatic properties of a novel αvβ3-binding disintegrin, DisBa-01, isolated from a cDNA library made with RNAs from the venom gland of Bothrops alternatus. The 11,637 Da-recombinant monomeric form of DisBa-01 displayed an RGD motif and interacted with purified αvβ3 integrin in surface plasmon resonance studies, in a dose-dependent and cation sensitive manner. A three-dimensional molecular model of DisBa-01 in complex with αvβ3 predicted a large surface of contacts with the β3 subunit. DisBa-01 inhibited the adhesion of αvβ3-expressing human microvascular endothelial cell line-1 (HMEC-1) and murine melanoma cell line B16F10 to vitronectin (IC50 = 555 nM and 225 nM, respectively), and transiently inhibited their proliferation without direct cell toxicity, but did not affect the binding nor the proliferation of a human breast cancer-derived cell line (MDA-MB-231) not expressing αvβ3. In vivo, DisBa-01 dose-dependently decreased bFGF-induced angiogenesis in a matrigel plug assay in athymic nude mice (IC50 = 83 nM). When injected intravenously to C57BL/6 mice together with B16F10 melanoma cells, DisBa-01 time- and dose-dependently inhibited lung metastasis monitored by bioluminescent imaging. We conclude that DisBa-01 is a potent new inhibitor of αvβ3-dependent adherence mechanisms involved in neo-vascularization and tumor metastasis processes.

Keywords

Disintegrinαvβ3AngiogenesisMetastasisBioluminescent imaging

Abbreviations

BFGF

Basic fibroblast growth factor

ECM

Extracellular matrix

FBS

Foetal bovine serum

FITC

Fluorescein isothiocyanate

GFP

Green fluorescent protein

HMEC-1

Human microvascular endothelial cell line-1

IPTG

Isopropyl thio-β-d-galactopyranoside

MD

Molecular dynamics

MIDAS

Metal-ion-dependent adhesion site

MS

Mass spectroscopy

PCR

Polymerase chain reaction

PDB

Protein data base

RGD

Arginine-glycine-aspartic acid

RU

Resonance units

SVMP

Snake venom metalloproteinase

Vn

Vitronectin

Copyright information

© Springer Science+Business Media B.V. 2007