A novel alpha(v)beta (3)-blocking disintegrin containing the RGD motive, DisBa-01, inhibits bFGF-induced angiogenesis and melanoma metastasis.
The integrin alpha(v)beta(3) 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 alpha(v)beta(3)-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 alpha(v)beta(3) 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 alpha(v)beta(3) predicted a large surface of contacts with the beta(3) subunit. DisBa-01 inhibited the adhesion of alpha(v)beta(3)-expressing human microvascular endothelial cell line-1 (HMEC-1) and murine melanoma cell line B16F10 to vitronectin (IC(50) = 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 alpha(v)beta(3). In vivo, DisBa-01 dose-dependently decreased bFGF-induced angiogenesis in a matrigel plug assay in athymic nude mice (IC(50) = 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 alpha(v)beta(3)-dependent adherence mechanisms involved in neo-vascularization and tumor metastasis processes.
Ramos
Oscar H P
OH
Dep. Ciências Fisiológicas, Universidade Federal de São Carlos, São Carlos, SP, Brasil.
Kauskot
Alexandre
A
Cominetti
Márcia R
MR
Bechyne
Iga
I
Salla Pontes
Carmen L
CL
Chareyre
Fabrice
F
Manent
Jan
J
Vassy
Roger
R
Giovannini
Marco
M
Legrand
Chantal
C
Selistre-de-Araujo
Heloisa S
HS
Crépin
Michel
M
Bonnefoy
Arnaud
A
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D001483:Base Sequence; D017837:Bothrops; D002448:Cell Adhesion / Q000187:drug effects; D003001:Cloning, Molecular; D003435:Crotalid Venoms / Q000737:chemistry / Q000494:pharmacology*; D019483:Disintegrins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D005346:Fibroblast Growth Factors / Q000378:metabolism; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000037:antagonists & inhibitors* / Q000187:drug effects; D008546:Melanoma, Experimental / Q000188:drug therapy*; D051379:Mice; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000188:drug therapy*; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
D000970:Antineoplastic Agents; D003435:Crotalid Venoms; D019483:Disintegrins; D039302:Integrin alphaVbeta3; D011994:Recombinant Proteins; D005346:Fibroblast Growth Factors