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Antiangiogenic activity of a neutralizing human single-chain antibody fragment against fibroblast growth factor receptor 1.
Mol Cancer Ther 2010; 9(12):3244-53MC

Abstract

Fibroblast growth factor receptor-1 (FGFR-1) transduces proangiogenic and proliferative signals in human cancers. Thus, FGFR-1 may represent a target for the development of antiangiogenic/antineoplastic therapies. We screened a human single-chain fragment variable (scFv) antibody phage display library against the extracellular domain of the FGFR-1-IIIc isoform that harbors the FGF binding site. Several phages were isolated and tested for specificity and sensitivity, and the most promising antibody fragment RR-C2 was characterized for its biochemical and biological properties. ScFv RR-C2 specifically recognizes FGFR-1α and FGFR-1β isoforms in ELISA, Western blotting, and surface plasmon resonance analysis with a K(d) value of 300 and 144 nmol/L for the 2 receptor isoforms, respectively. The antibody fragment also recognizes FGFR-1 when the receptor is exposed on the cell surface, thus preventing the formation of the ternary complex among FGFR-1, its ligand FGF2, and cell surface heparan sulfate proteoglycans. Accordingly, scFv RR-C2 specifically inhibits FGF2-mediated mitogenic activity in endothelial cells of human, bovine, and murine origin in a nanomolar range of concentrations. Also, the antibody fragment prevents FGF2-triggered sprouting of both human umbilical vein endothelial cell spheroids and of murine endothelium from aortic rings. Finally, the antibody fragment hampers the angiogenic activity exerted both by FGF2 in the chick embryo chorioallantoic membrane assay and by S115 mouse mammary tumor cells in the Matrigel plug assay. Taken together, the data show that scFv RR-C2 recognizes and neutralizes FGFR-1 activity in different animal species, including humans, thus representing a novel tool for the development of antiangiogenic/antineoplastic therapies.

Authors+Show Affiliations

Unit of General Pathology and Immunology, University of Brescia, Viale Europa 11, 25123 Brescia, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

20940322

Citation

Ronca, Roberto, et al. "Antiangiogenic Activity of a Neutralizing Human Single-chain Antibody Fragment Against Fibroblast Growth Factor Receptor 1." Molecular Cancer Therapeutics, vol. 9, no. 12, 2010, pp. 3244-53.
Ronca R, Benzoni P, Leali D, et al. Antiangiogenic activity of a neutralizing human single-chain antibody fragment against fibroblast growth factor receptor 1. Mol Cancer Ther. 2010;9(12):3244-53.
Ronca, R., Benzoni, P., Leali, D., Urbinati, C., Belleri, M., Corsini, M., ... Dell'Era, P. (2010). Antiangiogenic activity of a neutralizing human single-chain antibody fragment against fibroblast growth factor receptor 1. Molecular Cancer Therapeutics, 9(12), pp. 3244-53. doi:10.1158/1535-7163.MCT-10-0417.
Ronca R, et al. Antiangiogenic Activity of a Neutralizing Human Single-chain Antibody Fragment Against Fibroblast Growth Factor Receptor 1. Mol Cancer Ther. 2010;9(12):3244-53. PubMed PMID: 20940322.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Antiangiogenic activity of a neutralizing human single-chain antibody fragment against fibroblast growth factor receptor 1. AU - Ronca,Roberto, AU - Benzoni,Patrizia, AU - Leali,Daria, AU - Urbinati,Chiara, AU - Belleri,Mirella, AU - Corsini,Michela, AU - Alessi,Patrizia, AU - Coltrini,Daniela, AU - Calza,Stefano, AU - Presta,Marco, AU - Dell'Era,Patrizia, Y1 - 2010/10/12/ PY - 2010/10/14/entrez PY - 2010/10/14/pubmed PY - 2011/3/29/medline SP - 3244 EP - 53 JF - Molecular cancer therapeutics JO - Mol. Cancer Ther. VL - 9 IS - 12 N2 - Fibroblast growth factor receptor-1 (FGFR-1) transduces proangiogenic and proliferative signals in human cancers. Thus, FGFR-1 may represent a target for the development of antiangiogenic/antineoplastic therapies. We screened a human single-chain fragment variable (scFv) antibody phage display library against the extracellular domain of the FGFR-1-IIIc isoform that harbors the FGF binding site. Several phages were isolated and tested for specificity and sensitivity, and the most promising antibody fragment RR-C2 was characterized for its biochemical and biological properties. ScFv RR-C2 specifically recognizes FGFR-1α and FGFR-1β isoforms in ELISA, Western blotting, and surface plasmon resonance analysis with a K(d) value of 300 and 144 nmol/L for the 2 receptor isoforms, respectively. The antibody fragment also recognizes FGFR-1 when the receptor is exposed on the cell surface, thus preventing the formation of the ternary complex among FGFR-1, its ligand FGF2, and cell surface heparan sulfate proteoglycans. Accordingly, scFv RR-C2 specifically inhibits FGF2-mediated mitogenic activity in endothelial cells of human, bovine, and murine origin in a nanomolar range of concentrations. Also, the antibody fragment prevents FGF2-triggered sprouting of both human umbilical vein endothelial cell spheroids and of murine endothelium from aortic rings. Finally, the antibody fragment hampers the angiogenic activity exerted both by FGF2 in the chick embryo chorioallantoic membrane assay and by S115 mouse mammary tumor cells in the Matrigel plug assay. Taken together, the data show that scFv RR-C2 recognizes and neutralizes FGFR-1 activity in different animal species, including humans, thus representing a novel tool for the development of antiangiogenic/antineoplastic therapies. SN - 1538-8514 UR - https://www.unboundmedicine.com/medline/citation/20940322/Antiangiogenic_activity_of_a_neutralizing_human_single_chain_antibody_fragment_against_fibroblast_growth_factor_receptor_1_ L2 - http://mct.aacrjournals.org/cgi/pmidlookup?view=long&pmid=20940322 DB - PRIME DP - Unbound Medicine ER -