A urokinase receptor-derived peptide inhibiting VEGF-dependent directional migration and vascular sprouting.
The receptor for the urokinase-type plasminogen activator (uPAR) is a widely recognized master regulator of cell migration, and uPAR₈₈₋₉₂ is the minimal sequence required to induce cell motility. We previously showed that soluble forms of uPAR elicit angiogenic responses through their uPAR₈₈₋₉₂ chemotactic sequence and that the synthetic peptide SRSRY exerts similar effects. By a drug design approach, based on the conformational analysis of the uPAR₈₈₋₉₂ sequence, we developed peptides (pERERY, RERY, and RERF) that potently inhibit signaling triggered by uPAR₈₈₋₉₂. In this study, we present evidence that these peptides are endowed also with a clear-cut antiangiogenic activity, although to different extents. The most active, RERF, prevents tube formation by human endothelial cells exposed to SRSRY. RERF also inhibits VEGF-triggered endothelial cell migration and cord-like formation in a dose-dependent manner, starting in the femtomolar range. RERF prevents F-actin polymerization, recruitment of αvβ3 integrin at focal adhesions, and αvβ3/VEGFR2 complex formation in endothelial cells exposed to VEGF. At molecular level, the inhibitory effect of RERF on VEGF signaling is shown by the decreased amount of phospho-FAK and phospho-Akt in VEGF-treated cells. In vivo, RERF prevents VEGF-dependent capillary sprouts originating from the host vessels that invaded angioreactors implanted in mice and neovascularization induced by subcorneal implantation of pellets containing VEGF in rabbits. Consistently, RERF reduced the growth and vascularization rate of tumors formed by HT1080 cells injected subcutaneously in the flanks of nude mice, indicating that RERF is a promising therapeutic agent for the control of diseases fuelled by excessive angiogenesis such as cancer.
Corresponding Author: Maria Vincenza Carriero, Department of Experimental Oncology-National Cancer Institute of Naples, via M. Semmola, Naples 80131, Italy. m.carriero@istitutotumori.na.it.
Katia
K
Bifulco
Immacolata
I
Longanesi-Cattani
Eleonora
E
Liguori
Claudio
C
Arra
Domenica
D
Rea
Maria Teresa
MT
Masucci
Mario
M
De Rosa
Vincenzo
V
Pavone
Maria Patrizia
MP
Stoppelli
Maria Vincenza
MV
Carriero
D020533:Angiogenesis Inhibitors / Q000008:administration & dosage; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D060527:Corneal Keratocytes / Q000187:drug effects; D015195:Drug Design; D015972:Gene Expression Regulation, Neoplastic; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D010455:Peptides / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000737:chemistry; D011817:Rabbits; D055293:Receptors, Urokinase Plasminogen Activator / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics*; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D020533:Angiogenesis Inhibitors; D010455:Peptides; D055293:Receptors, Urokinase Plasminogen Activator; D042461:Vascular Endothelial Growth Factor A