Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
Charles T. Campbell Laboratory of Ophthalmic Microbiology, University of Pittsburgh, Pittsburgh, Pennsylvania.
J Ocul Pharmacol Ther. 2019 Nov;35(9):491-496. doi: 10.1089/jop.2019.0065. Epub 2019 Oct 10.
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. 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. All 4 domains bound to collagen I and IV , 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. 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.
泪膜和眨眼的联合活动在从眼表面清除异物方面非常有效。这阻止了某些类别的药物用于治疗眼表面问题。我们提出，使用与角膜上的部分结合的肽和蛋白质结构域可以通过将药物锚定在眼表面上足够长的时间来提供治疗效果，从而用于输送治疗剂。在这项研究中，我们评估了融合到细菌β-半乳糖苷酶中的 4 种不同的胶原结合结构域，以将报告蛋白递送到涂覆有 I 型和 IV 型胶原的平板、兔角膜和单纯疱疹病毒（HSV-1）感染的鼠角膜。所有 4 种结构域均与 I 型和 IV 型胶原结合，而仅来自牛血管性血友病因子（vWF）的 10 个氨基酸（AA）序列和来自细菌蛋白 ColH 的 215 个 AA 胶原结合结构域有效地与磨损的兔角膜结合。为了在临床相关模型中测试对角膜的结合，我们评估了 vWF 胶原结合肽融合物与 HSV-1 感染的鼠角膜的结合。我们观察到 vWF 衍生的肽介导与感染的角膜附着，而没有胶原结合结构域的报告蛋白则没有附着。向前推进，vWF 胶原结合肽可用作锚定物，以输送治疗剂，以防止因疾病和炎症导致的角膜表面受损而引起的瘢痕形成和视力丧失。
