ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF Signaling
Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) is the main mediator of angiogenic signaling in endothelial cells and a primary responder to VEGF. VEGF dependent VEGFR-2 activation regulates endothelial cell migration and proliferation, as well as vessel permeability. VEGF is presented as an...
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doaj-46dae29a4a8e447997888bcfe90f5d052020-11-25T00:49:16ZengMDPI AGInternational Journal of Molecular Sciences1422-00672018-05-01195133410.3390/ijms19051334ijms19051334ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF SignalingKurt Ballmer-Hofer0Caroline A.C. Hyde1Thomas Schleier2Dragana Avramovic3Laboratory of Biomolecular Research, Paul Scherrer Institut, 5232 Villigen, SwitzerlandLaboratory of Biomolecular Research, Paul Scherrer Institut, 5232 Villigen, SwitzerlandLaboratory of Biomolecular Research, Paul Scherrer Institut, 5232 Villigen, SwitzerlandLaboratory of Biomolecular Research, Paul Scherrer Institut, 5232 Villigen, SwitzerlandVascular Endothelial Growth Factor Receptor 2 (VEGFR-2) is the main mediator of angiogenic signaling in endothelial cells and a primary responder to VEGF. VEGF dependent VEGFR-2 activation regulates endothelial cell migration and proliferation, as well as vessel permeability. VEGF is presented as an antiparallel homodimer, and its binding to VEGFR-2 brings two receptors in close proximity. Downstream signaling is triggered by receptor dimerization, kinase activation, and receptor internalization. Our aim was to further investigate allosteric inhibition using binders targeting extracellular subdomains 4–7 of VEGFR-2 as an alternative to existing anti-angiogenic therapies, which rely on neutralizing VEGF or blocking of the ligand-binding site on the receptor. We applied phage display technology to produce single chain antibody fragments (scFvs) targeting VEGFR-2. Selected antibody fragments were characterized using biophysical and biological assays. We characterized several antibody fragments, which exert their inhibitory effect of VEGFR-2 independent of ligand binding. These reagents led to rapid clearance of VEGFR-2 from the cell surface without kinase activation, followed by an increase in intracellular receptor-positive vesicles, suggesting receptor internalization. Our highly specific VEGFR-2 binders thus represent novel tools for anti-angiogenic therapy and diagnostic applications.http://www.mdpi.com/1422-0067/19/5/1334angiogenesisVEGFR-2VEGFscFvinhibitionreceptor downregulation |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kurt Ballmer-Hofer Caroline A.C. Hyde Thomas Schleier Dragana Avramovic |
spellingShingle |
Kurt Ballmer-Hofer Caroline A.C. Hyde Thomas Schleier Dragana Avramovic ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF Signaling International Journal of Molecular Sciences angiogenesis VEGFR-2 VEGF scFv inhibition receptor downregulation |
author_facet |
Kurt Ballmer-Hofer Caroline A.C. Hyde Thomas Schleier Dragana Avramovic |
author_sort |
Kurt Ballmer-Hofer |
title |
ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF Signaling |
title_short |
ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF Signaling |
title_full |
ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF Signaling |
title_fullStr |
ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF Signaling |
title_full_unstemmed |
ScFvs as Allosteric Inhibitors of VEGFR-2: Novel Tools to Harness VEGF Signaling |
title_sort |
scfvs as allosteric inhibitors of vegfr-2: novel tools to harness vegf signaling |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2018-05-01 |
description |
Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) is the main mediator of angiogenic signaling in endothelial cells and a primary responder to VEGF. VEGF dependent VEGFR-2 activation regulates endothelial cell migration and proliferation, as well as vessel permeability. VEGF is presented as an antiparallel homodimer, and its binding to VEGFR-2 brings two receptors in close proximity. Downstream signaling is triggered by receptor dimerization, kinase activation, and receptor internalization. Our aim was to further investigate allosteric inhibition using binders targeting extracellular subdomains 4–7 of VEGFR-2 as an alternative to existing anti-angiogenic therapies, which rely on neutralizing VEGF or blocking of the ligand-binding site on the receptor. We applied phage display technology to produce single chain antibody fragments (scFvs) targeting VEGFR-2. Selected antibody fragments were characterized using biophysical and biological assays. We characterized several antibody fragments, which exert their inhibitory effect of VEGFR-2 independent of ligand binding. These reagents led to rapid clearance of VEGFR-2 from the cell surface without kinase activation, followed by an increase in intracellular receptor-positive vesicles, suggesting receptor internalization. Our highly specific VEGFR-2 binders thus represent novel tools for anti-angiogenic therapy and diagnostic applications. |
topic |
angiogenesis VEGFR-2 VEGF scFv inhibition receptor downregulation |
url |
http://www.mdpi.com/1422-0067/19/5/1334 |
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