title: The promotion of endothelial cell attachment and spreading using FNIII10 fused to VEGF-A165.
abstract: Synergy in the downstream signaling pathways of the vascular endothelial growth factor receptor-2 (VEGFR-2) and the integrin αvβ3 is critical for blood vessel formation. Thus, agents that activate both receptors could possess efficient pro-angiogenic potential. Here, we created a fibrin-binding bi-functional protein (FNIII10-VEGF) consisting of the 10th type III domain of fibronectin (FNIII10) fused to a plasmin-resistant VEGF-A165 mutant (VEGF) that potentiated angiogenic processes when compared to the effect of the separate molecules. FNIII10-VEGF was able to bind both VEGFR-2 and integrin αvβ3. Intriguingly, cell attachment and spreading to immobilized FNIII10-VEGF was significantly enhanced compared to individual FNIII10 or VEGF proteins. Delivery of immobilized FNIII10-VEGF by covalent linkage to a fibrin matrix significantly enhanced the angiogenic response in an in vivo wound healing assay compared to soluble VEGF. Unexpectedly, the angiogenic response to fibrin-immobilized FNIII10-VEGF was reduced in comparison to the pro-angiogenic effect of fibrin-immobilized VEGF. Collectively, findings of this study corroborate a critical role for a subtle balance of the integrin-VEGF interplay in angiogenesis and provide insight in how engineered growth factors in concert with biomaterial matrices may offer a potent molecular/material approach to harness these interactions for therapeutic angiogenesis.
authors.affiliation: Department of Dermatology, University of Cologne, Cologne, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Traub
authors.forename: Jessica
authors.initials: J
authors.lastname: Morgner
authors.forename: Mikaël M
authors.initials: MM
authors.lastname: Martino
authors.forename: Stefan
authors.initials: S
authors.lastname: Höning
authors.forename: Melody A
authors.initials: MA
authors.lastname: Swartz
authors.forename: Sara A
authors.initials: SA
authors.lastname: Wickström
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Hubbell
authors.forename: Sabine A
authors.initials: SA
authors.lastname: Eming
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D003921:Diabetes Mellitus, Experimental / Q000473:pathology; D004789:Enzyme Activation / Q000187:drug effects; D005337:Fibrin / Q000378:metabolism; D005353:Fibronectins / Q000737:chemistry* / Q000494:pharmacology*; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D008297:Male; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*; D012995:Solubility; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000378:metabolism
chemical_list: D005353:Fibronectins; D055767:Immobilized Proteins; D039302:Integrin alphaVbeta3; D011993:Recombinant Fusion Proteins; D042461:Vascular Endothelial Growth Factor A; D005337:Fibrin; D040301:Vascular Endothelial Growth Factor Receptor-2; D020830:rac1 GTP-Binding Protein