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Diabetologia

pp 1–13 | Cite as

Therapeutic regulation of VE-cadherin with a novel oligonucleotide drug for diabetic eye complications using retinopathy mouse models

  • Ka Ka Ting
  • Yang Zhao
  • Weiyong Shen
  • Paul Coleman
  • Michelle Yam
  • Tailoi Chan-Ling
  • Jia Li
  • Thorleif Moller
  • Mark Gillies
  • Mathew A. Vadas
  • Jennifer R. Gamble
Article

Abstract

Aims/hypothesis

A major feature of diabetic retinopathy is breakdown of the blood–retinal barrier, resulting in macular oedema. We have developed a novel oligonucleotide-based drug, CD5-2, that specifically increases expression of the key junctional protein involved in barrier integrity in endothelial cells, vascular-endothelial-specific cadherin (VE-cadherin). CD5-2 prevents the mRNA silencing by the pro-angiogenic microRNA, miR-27a. CD5-2 was evaluated in animal models of ocular neovascularisation and vascular leak to determine its potential efficacy for diabetic retinopathy.

Methods

CD5-2 was tested in three mouse models of retinal dysfunction: conditional Müller cell depletion, streptozotocin-induced diabetes and oxygen-induced retinopathy. Vascular permeability in the Müller cell-knockout model was assessed by fluorescein angiography. The Evans Blue leakage method was used to determine vascular permeability in streptozotocin- and oxygen-induced retinopathy models. The effects of CD5-2 on retinal neovascularisation, inter-endothelial junctions and pericyte coverage in streptozotocin- and oxygen-induced retinopathy models were determined by staining for isolectin-B4, VE-cadherin and neural/glial antigen 2 (NG2). Blockmir CD5-2 localisation in diseased retina was determined using fluorescent in situ hybridisation. The effects of CD5-2 on VE-cadherin expression and in diabetic retinopathy-associated pathways, such as the transforming growth factor beta (TGF-β) and wingless/integrated (WNT) pathway, were confirmed using western blot of lysates from HUVECs, a mouse brain endothelial cell line and a VE-cadherin null mouse endothelial cell line.

Results

CD5-2 penetrated the vasculature of the eye in the oxygen-induced retinopathy model. Treatment of diseased mice with CD5-2 resulted in reduced vascular leak in all three animal models, enhanced expression of VE-cadherin in the microvessels of the eye and improved pericyte coverage of the retinal vasculature in streptozotocin-induced diabetic models and oxygen-induced retinopathy models. Further, CD5-2 reduced the activation of retinal microglial cells in the streptozotocin-induced diabetic model. The positive effects of CD5-2 seen in vivo were further confirmed in vitro by increased protein expression of VE-cadherin, SMAD2/3 activity, and platelet-derived growth factor B (PDGF-B).

Conclusions/interpretation

CD5-2 has therapeutic potential for individuals with vascular-leak-associated retinal diseases based on its ease of delivery and its ability to reverse vascular dysfunction and inflammatory aspects in three animal models of retinopathy.

Keywords

Blood–retinal barrier Diabetic retinopathy Microglia MicroRNA Müller cells Neovascularisation Oligonucleotide Pericytes VE-cadherin 

Abbreviations

BBB

Blood–brain barrier

BRB

Blood–retinal barrier

DMO

Diabetic macular oedema

eNOS

Endothelial nitric oxide synthase

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

GCL

Ganglion cell layer

INL

Inner nuclear layer

KO

Knockout

LRP6

LDL receptor-related protein 6

miRNA

MicroRNA

NG2

Neural/glial antigen 2

OIR

Oxygen-induced retinopathy

ONL

Outer nuclear layer

P7/12/14/17

7/12/14/17 days postnatal

PDGF-B

Platelet-derived growth factor B

PDGFR-β

Platelet-derived growth factor receptor β

PDR

Proliferative diabetic retinopathy

SMAD

Smad family member

STZ

Streptozotocin

VE-cadherin

Vascular-endothelial-specific cadherin

VEC-null

VE-cadherin null

VEGF

Vascular endothelial growth factor

WNT

Wingless/integrated

Notes

Acknowledgements

We give special thanks to J. Hunter (Centenary Institute, Sydney, Australia) for preparing the human endothelial cells and to the imaging and animal facility staff at the Centenary Institute for their technical assistance. We thank E. Dejana (Italian Foundation for Cancer Research [FIRC], Institute of Molecular Oncology [IFOM], Milan, Italy) for supplying the VEC-null endothelial cells and G. Grau (Department of Pathology, Sydney Medical School, University of Sydney, Sydney, NSW, Australia) for the mouse brain endothelial cells.

Contribution statement

KKT designed the study, acquired and analysed the data, drafted and approved the final version of the manuscript. JRG and MAV contributed to the conception of the study, and drafted and approved the final version of the manuscript. MG and TM contributed to the conception and design of the study. TC-L was involved in interpretation of the data. WYS, MY, YZ, JL and PC acquired data, revised the article’s intellectual content and approved the final version. MG, TM and TC-L revised the article’s intellectual content and approved final version. KKT had full access to all the data, excluding those relating to the Müller cell transgenic model in this study. KKT and JRG are responsible for the integrity of this study.

Funding

This research was supported by grants from the National Health and Medical Research Council (NHMRC) of Australia (no. 571408). JRG holds the Wenkart Chair of the Endothelium at the Centenary Institute and the Sydney Medical School, University of Sydney.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2018_4770_MOESM1_ESM.pdf (15.9 mb)
ESM (PDF 16291 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Ka Ka Ting
    • 1
  • Yang Zhao
    • 1
  • Weiyong Shen
    • 2
  • Paul Coleman
    • 1
  • Michelle Yam
    • 2
  • Tailoi Chan-Ling
    • 3
  • Jia Li
    • 1
  • Thorleif Moller
    • 4
  • Mark Gillies
    • 2
  • Mathew A. Vadas
    • 1
  • Jennifer R. Gamble
    • 1
  1. 1.Centre for the Endothelium Vascular Biology Program Centenary InstituteThe University of SydneyNewtownAustralia
  2. 2.Save Sight InstituteThe University of SydneySydneyAustralia
  3. 3.Discipline of Anatomy and Histology, School of Medical Sciences Bosch InstituteThe University of SydneySydneyAustralia
  4. 4.Ranger Biotechnologies A/SÅrslevDenmark

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