title: Structure-based design of an urokinase-type plasminogen activator receptor-derived peptide inhibiting cell migration and lung metastasis.
abstract: The urokinase-type plasminogen activator receptor (uPAR) plays a central role in sustaining the malignant phenotype and promoting tumor metastasis. The Ser(88)-Arg-Ser-Arg-Tyr(92) is the minimum chemotactic sequence of uPAR required to induce the same intracellular signaling as its ligand uPA. Here, we describe the generation of new peptide inhibitors of cell migration and invasion derived from SRSRY by a drug design approach. Ac-Arg-Glu-Arg-Phe-NH(2) (i.e., RERF), which adopts a turned structure in solution, was selected for its ability to potently prevent SRSRY-directed cell migration. Fluorescein-RERF associates with very high affinity to RBL-2H3 rat basophilic leukemia cells expressing the human formyl peptide receptor (FPR). Accordingly, femtomolar concentrations of RERF prevent agonist-dependent internalization of FPR and inhibit N-formyl-Met-Leu-Phe-dependent migration in a dose-dependent manner. In the absence of FPR, fluorescein-RERF binds to cell surface at picomolar concentrations in an alphav integrin-dependent manner. The involvement of vitronectin receptor is further supported by the findings that 100 pmol/L RERF selectively inhibits vitronectin-dependent RBL-2H3 cell migration and prevents SRSRY-triggered uPAR/alphav association. Furthermore, RERF reduces the speed of wound closure and the extent of Matrigel invasion by human fibrosarcoma HT1080 cells without affecting cell proliferation. Finally, a 3- to 5-fold reduction of lung metastasis number and size in nude mice following i.v. injection of green fluorescent protein-expressing HT1080 cells in the presence of 3.32 mg/kg RERF is observed. Our findings indicate that RERF effectively prevents malignant cell invasion in vivo with no signs of toxicity and may represent a promising prototype drug for anticancer therapy.
authors.affiliation: Department of Experimental Oncology, National Cancer Institute of Naples, Via M. Semmola, 80131 Naples, Italy. mariolinacarriero@yahoo.it
authors.forename: Maria Vincenza
authors.initials: MV
authors.lastname: Carriero
authors.forename: Immacolata
authors.initials: I
authors.lastname: Longanesi-Cattani
authors.forename: Katia
authors.initials: K
authors.lastname: Bifulco
authors.forename: Ornella
authors.initials: O
authors.lastname: Maglio
authors.forename: Liliana
authors.initials: L
authors.lastname: Lista
authors.forename: Antonio
authors.initials: A
authors.lastname: Barbieri
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Votta
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Masucci
authors.forename: Claudio
authors.initials: C
authors.lastname: Arra
authors.forename: Renato
authors.initials: R
authors.lastname: Franco
authors.forename: Mario
authors.initials: M
authors.lastname: De Rosa
authors.forename: Maria Patrizia
authors.initials: MP
authors.lastname: Stoppelli
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Pavone
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000187:drug effects*; D005260:Female; D005354:Fibrosarcoma / Q000473:pathology; D006801:Humans; D047468:Immunoprecipitation; D008175:Lung Neoplasms / Q000556:secondary*; D051379:Mice; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D009362:Neoplasm Metastasis / Q000517:prevention & control*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000494:pharmacology*; D011487:Protein Conformation; D051381:Rats; D055293:Receptors, Urokinase Plasminogen Activator / Q000737:chemistry*
chemical_list: D010446:Peptide Fragments; D055293:Receptors, Urokinase Plasminogen Activator