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COMP-Ang1 stabilizes hyperglycemic disruption of blood-retinal barrier phenotype in human retinal microvascular endothelial cells

Rochfort, Keith D. ORCID: 0000-0003-1198-5952, Carroll, Lara S., Barabas, Peter ORCID: 0000-0002-8166-3476, Curtis, Timothy M. ORCID: 0000-0003-1543-2781, Ambati, Balamurali, Barron, Niall ORCID: 0000-0003-4602-326X and Cummins, Phil (2019) COMP-Ang1 stabilizes hyperglycemic disruption of blood-retinal barrier phenotype in human retinal microvascular endothelial cells. Investigative Ophthalmology & Visual Science, 60 (10). pp. 3547-3555. ISSN 1552-5783

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Abstract

PURPOSE. Current treatments for diabetic retinopathy (DR) have considerable limitations, underpinning the need for new therapeutic options. In this article, the ability of an engineered angiopoietin-1 variant (COMP-Ang1) to ameliorate the injurious effects of hyperglycemia on barrier integrity in a human retinal microvascular endothelial cell (HRMvEC) model is comprehensively investigated. METHODS. Confluent HRMvECs were treated (0–72 hours) with D-glucose (5 or 30 mM) in the absence and presence of COMP-Ang1 (10–200 ng/mL). L-glucose (30 mM) was used as osmotic control. Posttreatment, intact cell monolayers were monitored for permeability to FITC-dextran 40 kDa. Cells were also harvested for analysis of interendothelial junction targets by RT-qPCR and Western blotting. The impact of receptor tyrosine kinase Tie2 gene silencing on COMP-Ang1 efficacy was also evaluated. RESULTS. Treatment with 30 mM D-glucose (but not L-glucose) demonstrated a time-dependent elevation in the mean rate of FITC-dextran diffusion across intact HRMvEC monolayers, in parallel with significant reductions in mRNA/protein levels of occludin, claudin-5, ZO-1, and VE-Cadherin. These effects were all attenuated by COMP-Ang1 in a concentration-dependent fashion, with 200 ng/mL recovering barrier function by ~88%, and recovering reduced interendothelial junction protein levels by more than 50%. Finally, Tie2 knockdown by small interfering RNA silencing blocked the ability of COMP-Ang1 to mitigate against hyperglycemia-induced permeabilization of HRMvECs and depletion of junctional expression levels. CONCLUSIONS. In summary, this article presents a reproducible in vitro cell study that quantifies the concentration-dependent efficacy of COMP-Ang1 to mitigate the injurious effects of hyperglycemic challenge on HRMvEC barrier properties via Tie2-mediated signaling.

Item Type:Article (Published)
Refereed:Yes
Uncontrolled Keywords:endothelial; diabetic retinopathy; COMP-Ang1; Tie2; blood-retinal barrier
Subjects:Biological Sciences > Biotechnology
Biological Sciences > Cell biology
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Publisher:Association for Research in Vision and Ophthalmology (ARVO)
Official URL:https://dx.doi.org/10.1167/iovs.19-27644
Copyright Information:© 2019 The Authors. Open Access (CC BY-NC-ND 4.0)
ID Code:27863
Deposited On:14 Oct 2022 12:44 by Thomas Murtagh . Last Modified 14 Oct 2022 13:13

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