Dissecting the role of matrix metalloproteinases (MMP) and integrin alpha(v)beta3 in angiogenesis in vitro: absence of hemopexin C domain bioactivity, but membrane-Type 1-MMP and alpha(v)beta3 are critical

Matrix metalloproteinase (MMP)-2 and its hemopexin C domain autolytic fragment (also called PEX) have been proposed to be crucial for angiogenesis. Here, we have investigated the dependency of in vitro angiogenesis on MMP-mediated extracellular proteolysis and integrin alpha(v)beta3-mediated cell ad...

Full description

Saved in:
Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 2005-10, Vol.65 (20), p.9377-9387
Main Authors: Nisato, Riccardo E, Hosseini, Ghamartaj, Sirrenberg, Christian, Butler, Georgina S, Crabbe, Thomas, Docherty, Andrew J P, Wiesner, Matthias, Murphy, Gillian, Overall, Christopher M, Goodman, Simon L, Pepper, Michael S
Format: Article
Language:English
Subjects:
Animals
Cattle
Cytokines - pharmacology
Endothelial Cells - enzymology
Endothelial Cells - metabolism
Enzyme Activation
Fibroblast Growth Factor 2 - pharmacology
Hemopexin - metabolism
Hemopexin - pharmacology
Humans
Integrin alphaVbeta3 - metabolism
Matrix Metalloproteinase 2 - biosynthesis
Matrix Metalloproteinase 2 - genetics
Matrix Metalloproteinase 2 - metabolism
Matrix Metalloproteinase 2 - pharmacology
Matrix Metalloproteinases, Membrane-Associated
Metalloendopeptidases - antagonists & inhibitors
Metalloendopeptidases - biosynthesis
Metalloendopeptidases - metabolism
Neovascularization, Physiologic - physiology
Oligopeptides - pharmacology
Protein Structure, Tertiary
RNA, Messenger - biosynthesis
RNA, Messenger - genetics
Vascular Endothelial Growth Factor A - pharmacology
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Matrix metalloproteinase (MMP)-2 and its hemopexin C domain autolytic fragment (also called PEX) have been proposed to be crucial for angiogenesis. Here, we have investigated the dependency of in vitro angiogenesis on MMP-mediated extracellular proteolysis and integrin alpha(v)beta3-mediated cell adhesion in a three-dimensional collagen I model. The hydroxamate-based synthetic inhibitors BB94, CT1399, and CT1847 inhibited endothelial cell invasion, as did neutralizing anti-membrane-type 1-MMP (MT1-MMP) antibodies and tissue inhibitor of MMP (TIMP)-2 and TIMP-3 but not TIMP-1. This confirmed the pivotal importance of MT1-MMP over other MMPs in this model. Invasion was also inhibited by a nonpeptidic antagonist of integrin alpha(v)beta3, EMD 361276. Although PEX strongly inhibited pro-MMP-2 activation, when contaminating lipopolysaccharide was neutralized, PEX neither affected angiogenesis nor bound integrin alpha(v)beta(3). Moreover, no specific binding of pro-MMP-2 to integrin alpha(v)beta3 was found, whereas only one out of four independently prepared enzymatically active MMP-2 preparations could bind integrin alpha(v)beta3 , and this in a PEX-independent manner. Likewise, integrin alpha(v)beta3 -expressing cells did not bind MMP-2-coated surfaces. Hence, these findings show that endothelial cell invasion of collagen I gels is MT1-MMP and alpha(v)beta3 - dependent but MMP-2 independent and does not support a role for PEX in alpha(v)beta3 integrin binding or in modulating angiogenesis in this system.
ISSN:0008-5472
1538-7445