Jes K Klarlund1, Jake D Callaghan1, Nicholas A Stella1,2, Regis P Kowalski1,2, Nancy A McNamara3,4, Robert M Q Shanks1,2. 1. Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania. 2. Charles T. Campbell Laboratory of Ophthalmic Microbiology, University of Pittsburgh, Pittsburgh, Pennsylvania. 3. School of Optometry, University of California, Berkeley, Berkeley, California. 4. Department of Anatomy, University of California, San Francisco, San Francisco, California.
Abstract
Purpose: The combined activity of the tear film and blinking is remarkably efficient at removal of foreign materials from the ocular surface. This has prevented the use of certain classes of drugs for the treatment of ocular surface problems. We propose that the use of peptide and protein domains that bind to moieties on the cornea could be used to deliver therapeutics by anchoring the drugs on the ocular surface long enough to provide therapeutic effects. Methods: In this study, we evaluated 4 different collagen binding domains fused to bacterial β-galactosidase for delivery of a reporter protein to collagen I and collagen IV-coated plates, rabbit corneas, and Herpes simplex virus (HSV-1) infected mouse corneas. Results: All 4 domains bound to collagen I and IV in vitro, whereas only a 10 amino acid (AA) sequence from bovine von Willebrand factor (vWF) and a 215 AA collagen binding domain from the bacterial protein ColH efficiently bound to abraded rabbit corneas. To test binding to corneas in a clinically relevant model, we assessed binding of the vWF collagen binding peptide fusions to HSV-1 infected mouse corneas. We observed that the vWF derived peptide mediated attachment to infected corneas, whereas the reporter protein without a collagen binding domain did not bind. Conclusions: Moving forward, the vWF collagen binding peptide could be used as an anchor to deliver therapeutics to prevent scarring and vision loss from damaged corneal surfaces due to disease and inflammation.
Purpose: The combined activity of the tear film and blinking is remarkably efficient at removal of foreign materials from the ocular surface. This has prevented the use of certain classes of drugs for the treatment of ocular surface problems. We propose that the use of peptide and protein domains that bind to moieties on the cornea could be used to deliver therapeutics by anchoring the drugs on the ocular surface long enough to provide therapeutic effects. Methods: In this study, we evaluated 4 different collagen binding domains fused to bacterial β-galactosidase for delivery of a reporter protein to collagen I and collagen IV-coated plates, rabbit corneas, and Herpes simplex virus (HSV-1)infectedmouse corneas. Results: All 4 domains bound to collagen I and IV in vitro, whereas only a 10 amino acid (AA) sequence from bovinevon Willebrand factor (vWF) and a 215 AA collagen binding domain from the bacterial protein ColH efficiently bound to abraded rabbit corneas. To test binding to corneas in a clinically relevant model, we assessed binding of the vWF collagen binding peptide fusions to HSV-1 infectedmouse corneas. We observed that the vWF derived peptide mediated attachment to infected corneas, whereas the reporter protein without a collagen binding domain did not bind. Conclusions: Moving forward, the vWF collagen binding peptide could be used as an anchor to deliver therapeutics to prevent scarring and vision loss from damaged corneal surfaces due to disease and inflammation.
Authors: Carla S Medeiros; Gustavo K Marino; Marcony R Santhiago; Steven E Wilson Journal: Invest Ophthalmol Vis Sci Date: 2018-08-01 Impact factor: 4.799