Engineered ligand-based VEGFR antagonists with increased receptor binding affinity more effectively inhibit angiogenesis.

Bioengineering & Translational Medicine
Shiven KapurJennifer R Cochran

Abstract

Pathologic angiogenesis is mediated by the coordinated action of the vascular endothelial growth factor (VEGF)/vascular endothelial growth factor receptor 2 (VEGFR2) signaling axis, along with crosstalk contributed by other receptors, notably αvβ3 integrin. We build on earlier work demonstrating that point mutations can be introduced into the homodimeric VEGF ligand to convert it into an antagonist through disruption of binding to one copy of VEGFR2. This inhibitor has limited potency, however, due to loss of avidity effects from bivalent VEGFR2 binding. Here, we used yeast surface display to engineer a variant with VEGFR2 binding affinity approximately 40-fold higher than the parental antagonist, and 14-fold higher than the natural bivalent VEGF ligand. Increased VEGFR2 binding affinity correlated with the ability to more effectively inhibit VEGF-mediated signaling, both in vitro and in vivo, as measured using VEGFR2 phosphorylation and Matrigel implantation assays. High affinity mutations found in this variant were then incorporated into a dual-specific antagonist that we previously designed to simultaneously bind to and inhibit VEGFR2 and αvβ3 integrin. The resulting dual-specific protein bound to human and murine endothelia...Continue Reading

References

Feb 4, 1997·Proceedings of the National Academy of Sciences of the United States of America·J L ArbiserJ Folkman
Jun 1, 1997·Nature Biotechnology·E T Boder, K D Wittrup
May 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·G SiemeisterG Martiny-Baron
Aug 2, 2002·The Journal of Biological Chemistry·Thomas P BoesenTorben Halkier
Aug 15, 2002·Proceedings of the National Academy of Sciences of the United States of America·Jocelyn HolashJohn S Rudge
Aug 15, 2002·Proceedings of the National Academy of Sciences of the United States of America·Eugene S KimDarrell J Yamashiro
Jun 5, 2003·Nature Medicine·Napoleone FerraraJennifer LeCouter
May 25, 2004·Expert Opinion on Pharmacotherapy·Shikha Nanda, Joan M Bathon
Aug 4, 2004·Methods in Enzymology·David W ColbyK Dane Wittrup
Aug 6, 2004·Endocrine Reviews·Napoleone Ferrara
Oct 13, 2006·The Journal of Experimental Medicine·Ganapati H MahabeleshwarTatiana V Byzova
Jan 11, 2007·Proceedings of the National Academy of Sciences of the United States of America·Michael I DorrellMartin Friedlander
Apr 5, 2007·Nature Protocols·Ginger ChaoK Dane Wittrup
Jul 21, 2007·Circulation Research·Ganapati H MahabeleshwarTatiana V Byzova
Oct 20, 2007·Nature Protocols·Michael S RogersRobert J D'Amato
Feb 22, 2008·Cell Cycle·Ganapati H MahabeleshwarTatiana V Byzova
Jul 25, 2008·Nature Reviews. Cancer·Gabriele Bergers, Douglas Hanahan
Oct 2, 2008·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Junhang ZhangEric Wahlberg
Dec 3, 2008·Cell Biochemistry and Biophysics·Payaningal R SomanathTatiana V Byzova
Mar 10, 2009·Angiogenesis·Payaningal R SomanathTatiana V Byzova
Nov 3, 2009·European Journal of Ophthalmology·Hailton B OliveiraJin-Hong Chang
May 8, 2010·Trends in Biotechnology·Angel M CuestaLuis Alvarez-Vallina
Dec 15, 2010·Journal of Angiogenesis Research·Domenico Ribatti
Jun 3, 2011·The Journal of Clinical Investigation·Masahiro MurakamiMichael Simons
Aug 10, 2011·Proceedings of the National Academy of Sciences of the United States of America·Niv PapoJennifer R Cochran
Apr 17, 2013·Current Opinion in Biotechnology·Mihalis S KariolisJennifer R Cochran
Sep 5, 2013·Cold Spring Harbor Perspectives in Biology·Michael JeltschKari Alitalo
Jun 21, 2014·Cell Death and Differentiation·J LemkeH Walczak
Sep 23, 2014·Nature Chemical Biology·Mihalis S KariolisJennifer R Cochran
Feb 1, 2015·Molecular Immunology·Christoph SpiessPaul J Carter

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Citations

Jan 10, 2018·Bioengineering & Translational Medicine·Pankaj Karande

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Methods Mentioned

BETA
PCR
flow cytometry
circular dichroism
size exclusion chromatography
protein assay

Software Mentioned

FlowJo
Zen Blue
KaleidaGraph
CellQuest

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