Structure-Guided Molecular Engineering of a Vascular Endothelial Growth Factor Antagonist to Treat Retinal Diseases
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Cellular and Molecular Bioengineering ( 2020) https://doi.org/10.1007/s12195-020-00641-0
2020 CMBE Young Innovators issue
Structure-Guided Molecular Engineering of a Vascular Endothelial Growth Factor Antagonist to Treat Retinal Diseases RAKEEB KURESHI,1 ANGELA ZHU,2 JIKUI SHEN,3,4 STEPHANY Y. TZENG,1,5,6 LEILANI R. ASTRAB,1,2 PAUL R. SARGUNAS,2 JORDAN J. GREEN,1,2,3,5,6,7,8 PETER A. CAMPOCHIARO,3,4 and JAMIE B. SPANGLER 1,2,5 1
Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 2Department of Chemical & Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA; 3Department of Ophthalmology, The Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 4Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 5Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 6Insititute for Nanobiotechnology, Johns Hopkins University, Baltimore, MD, USA; 7Department of Materials Science & Engineering, Johns Hopkins University, Baltimore, MD, USA; and 8 Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA (Received 14 February 2020; accepted 22 July 2020) Associate Editor Scott Simon oversaw the review of this article.
Abstract Background—Ocular neovascularization is a hallmark of retinal diseases including neovascular age-related macular degeneration and diabetic retinopathy, two leading causes of blindness in adults. Neovascularization is driven by the interaction of soluble vascular endothelial growth factor (VEGF) ligands with transmembrane VEGF receptors (VEGFR), and inhibition of the VEGF pathway has shown tremendous clinical promise. However, anti-VEGF therapies require invasive intravitreal injections at frequent intervals and high doses, and many patients show incomplete responses to current drugs due to the lack of sustained VEGF signaling suppression. Methods—We synthesized insights from structural biology with molecular engineering technologies to engineer an antiVEGF antagonist protein. Starting from the clinically approved decoy receptor protein aflibercept, we strategically designed a yeast-displayed mutagenic library of variants and isolated clones with superior VEGF affinity compared to the clinical drug. Our lead engineered protein was expressed in the choroidal space of rat eyes via nonviral gene delivery. Results—Using a structure-informed directed evolution approach, we identified multiple promising anti-VEGF antagonist proteins with improved target affinity. Improvements were primarily mediated through reduction in dissociation rate, and structurally significant convergent sequence mutations were identified. Nonviral gene transfer of our engineered antagonist protein demonstrated robust and
Address correspondence to Jamie B. Spangler, Department of Chemical & Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA. Electronic mail: [email protected]
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