title: Design of a variant of vascular endothelial growth factor-A (VEGF-A) antagonizing KDR/Flk-1 and Flt-1.
abstract: Because of its central role in pathological angiogenesis, vascular endothelial growth factor (VEGF) has become a major target for anti-angiogenic therapies. We report here the construction of a heterodimeric antagonistic VEGF variant (HD-VEGF). In this antagonist, binding domains for the VEGF-receptors KDR/Flk-1 and Flt-1 are present at one pole of the dimer, whereas the other pole carries domain swap mutations, which prevent binding to either receptor. As HD-VEGF can only bind to monomeric receptors, it does not lead to signal transduction. Moreover, it antagonizes VEGF and possibly other members of the VEGF family, which are KDR/Flk-1 and Flt-1 ligands. We show here that HD-VEGF is a potent inhibitor of VEGF-mediated proliferation and tissue factor induction in endothelial cell cultures, requiring only a 20-fold and a 4-fold excess, respectively, to block the activity of wtVEGF completely. A 4-fold excess of HD-VEGF over wtVEGF was also sufficient to abrogate vascular permeability as determined in the Miles assay in vivo. Furthermore, HD-VEGF inhibited fetal bone angiogenesis in an ex vivo assay. Thus, HD-VEGF blocks KDR- and Flt-1-mediated VEGF activities that are crucial in the angiogenic process and is therefore a promising, multipotent compound in the treatment of angiogenesis-related diseases.
authors.affiliation: Department of Pathology, University Medical Center Nijmegen, The Netherlands. w.leenders@pathol.azn.nl
authors.forename: William
authors.initials: W
authors.lastname: Leenders
authors.forename: Nicolette
authors.initials: N
authors.lastname: Lubsen
authors.forename: Margarethe
authors.initials: M
authors.lastname: van Altena
authors.forename: Matthias
authors.initials: M
authors.lastname: Clauss
authors.forename: Martine
authors.initials: M
authors.lastname: Deckers
authors.forename: Clemens
authors.initials: C
authors.lastname: Löwik
authors.forename: Georg
authors.initials: G
authors.lastname: Breier
authors.forename: Dirk
authors.initials: D
authors.lastname: Ruiter
authors.forename: Robert
authors.initials: R
authors.lastname: de Waal
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005314:Embryonic and Fetal Development; D016228:Endothelial Growth Factors / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006168:Guinea Pigs; D008682:Metatarsal Bones / Q000098:blood supply / Q000187:drug effects / Q000196:embryology; D051379:Mice; D008969:Molecular Sequence Data; D018995:Myosin Heavy Chains; D018919:Neovascularization, Physiologic / Q000187:drug effects; D024501:Nonmuscle Myosin Type IIB; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000037:antagonists & inhibitors* / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor; D042461:Vascular Endothelial Growth Factor A; D040281:Vascular Endothelial Growth Factor Receptor-1
chemical_list: D016228:Endothelial Growth Factors; D016326:Extracellular Matrix Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D017978:Receptors, Growth Factor; D042461:Vascular Endothelial Growth Factor A; C501163:Flt1 protein, mouse; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor; D040281:Vascular Endothelial Growth Factor Receptor-1; D024501:Nonmuscle Myosin Type IIB; C103815:nonmuscle myosin type IIB heavy chain; D018995:Myosin Heavy Chains