Suppression of angiogenesis by lentiviral delivery of PEX, a noncatalytic fragment of matrix metalloproteinase 2

Abstract

Modulation of the balance between pro-and antiangiogenic factors holds great promise for the treatment of a broad spectrum of human disease ranging from ischemic heart disease to cancer. This requires both the identification of angiogenic regulators and their efficient delivery to target organs. Here, we demonstrate the use of a noncatalytic fragment of matrix metalloproteinase 2 (termed PEX) delivered by lentiviral vectors in different angiogenesis models. Transduction of human endothelial cells with PEX virus suppressed endothelial invasion and formation of capillary-like structures without affecting chemotaxis in vitro. Lentiviral delivery of PEX blocked basic fibroblast growth factor-induced matrix metalloproteinase 2 activation and angiogenesis on chicken chorioallantoic membranes. PEX expression also inhibited tumor-induced angiogenesis and tumor growth in a nude mouse model. Thus, our study shows that lentiviral vectors can deliver sufficient quantities of antiangiogenic substances to achieve therapeutic effects in vivo.

Full-length ADAMTS-1 and the ADAMTS-1 fragments display pro- and antimetastatic activity, respectively

“…These results are consistent with recent studies in cancer research that have demonstrated the pro-and antitumor activity, respectively, of particular intact proteins and their proteolytic fragments (O'Reilly et al, 1999;Pfeifer et al, 2000;Yi and Ruoslahti, 2001;Maeshima et al, 2002). How can auto-proteolytic cleavage convert a protumor factor into an antitumor agent?…”

A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA

“…Lentiviral Vector Production. Recombinant lentiviruses were produced by transient transfection in 293T cells using the calciumphosphate method as described (7,9,10). Infectious lentiviruses were harvested at 48 and 72 h posttransfection and filtered through 0.22-m-pore cellulose acetate filters as described (7,9,10).…”

Gelatinase-mediated migration and invasion of cancer cells

“…In some models, the cell surface activity of MMP-2 was found to be dependent on the α V β 3 integrin interaction and this interaction was necessary for tumor angiogenesis (Brooks et al, 1998;Brooks et al, 1996). Delivery of the MMP-2 C-terminal domain as a recombinant protein or via viral infection also potently suppressed angiogenesis (Pfeifer et al, 2000). The C-terminal domain of MMP-2 appears to be a naturally occuring proteolytic fragment and an inhibitor of pericellular MMP-2 activity (Bello et al, 2001;Brooks et al, 1998).…”