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A phage display-based approach to investigate abnormal neovessels of the retina

Staniszewska, Magdalena and Gu, Xiaolin and Romano, Carmelo and Kazlauskas, Andrius (2012) A phage display-based approach to investigate abnormal neovessels of the retina. Investigative Ophthalmalogy & Visual Sciences, 53 (8). pp. 4371-4379. ISSN 0146-0404

Abstract

PURPOSE. The goal of this project was to identify markers of abnormal neovascularization of the retinal vasculature, which is quintessential of pathological angiogenesis that occurs in blinding diseases such as proliferative diabetic retinopathy.
METHODS. Abnormal retinal neovascularization was induced in rat pups by subjecting them to the 50/10 oxygen-induced retinopathy (OIR) protocol, which involves fluctuating levels of ambient oxygen. A peptide library (that was displayed on phage) was positively and negatively screened over the surface of retinas isolated from experimental and control rats, respectively. Binding of phage to retinal vessels was evaluated by confocal microscopy of retinal flat mounts decorated with fluorescently labeled phage. The topography of the inner limiting membrane was studied by scanning electron microscopy.
RESULTS. Screening a library of peptides displayed on phage over the surface of OIR retinas resulted in isolation of a particular phage (SH phage) that distinguished between abnormal neovessels and the normal vasculature. As expected, the recognition of abnormal neovessels relied on the unique peptide insert of SH phage. Abnormal neovessels consisted of at least 3 cell types that were present in the following order of descending abundance: endothelial>pericytes>macrophage/microglia. SH phage recognized both endothelial cells and macrophage/microglia. Finally, SH phage decorated abnormal neovessels at an early stage of their genesis.
CONCLUSIONS. Abnormal development of neovessels is associated with early expression of distinct epitopes on the surface of cells within the pathological vasculature. Screening phage display libraries is one approach to detect such changes, and the resulting phage are potential imaging tools and/or drug delivery vehicles.

Item Type: Article
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Additional Information: This project was a research collaboration between investigators at Schepens and Alcon, supported by Alcon. Phage display was used to identify sequences that would bind specifically to pre-retinal neovascular tufts in OIR model retinas, and not to control retinas. The success in identifying such a phage is described in this manuscript. Alcon filed a patent on the phage and sequence. However, after more than a two years of trying to convert the identified sequence into a LMW agent displaying high affinity and specificity for pathological neovessels, the effort was abandoned as low likelihood of succeeding. Ownership of and responsibility for the patent was transferred to Schepens.
Keywords: neovascularization; retina; diabetes; retinopathy; phage display; oxygen-induced retinopathy
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Date Deposited: 13 Oct 2015 13:14
Last Modified: 13 Oct 2015 13:14
URI: https://oak.novartis.com/id/eprint/6600

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