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Nucleoside diphosphate kinase B deficiency causes a diabetes-like vascular pathology via up-regulation of endothelial angiopoietin-2 in the retina

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Abstract

Aims

Nucleoside diphosphate kinase B (NDPKB) is capable of maintaining the cellular nucleotide triphosphate pools. It might therefore supply UTP for the formation of UDP-GlcNAc from glucose. As NDPKB contributes to vascular dysfunction, we speculate that NDPKB might play a role in microangiopathies, such as diabetic retinopathy (DR). Therefore, we investigated the impact of NDPKB on retinal vascular damage using NDPKB−/− mice during development of DR and its possible mechanisms.

Methods

Pericyte loss and acellular capillary (AC) formation were assessed in streptozotocin-induced diabetic NDPKB−/− and wild-type (WT) mice. Expression of angiopoietin-2 (Ang2) and protein N-acetylglucosamine modification (GlcNAcylation) were assessed by western blot and/or immunofluorescence in the diabetic retinas as well as in endothelial cells depleted of NDPKB by siRNA and stimulated with high glucose.

Results

Similar to diabetic WT retinas, non-diabetic NDPKB−/− retinas showed a significant decrease in pericyte coverage in comparison with non-diabetic WT retinas. Hyperglycemia further aggravates pericyte loss in diabetic NDPKB−/− retinas. AC formation was detected in the diabetic NDPKB−/− retinas. Similar to hyperglycemia, NDPKB deficiency induced Ang2 expression and protein GlcNAcylation that were not further altered in the diabetic retinas. In cultured endothelial cells, stimulation with high glucose and NDPKB depletion comparably increased Ang2 expression and protein GlcNAcylation.

Conclusions

Our data identify NDPKB as a protective factor in the retina, which controls Ang2 expression and the hexosamine pathway. NDPKB-deficient mice are a suitable model for studying mechanisms underlying diabetic retinal vascular damage.

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Acknowledgments

The authors thank Doris Baltus, Heike Rauscher and Kristina Stephan-Schnatz for excellent technical support. This study was supported by grants from the European Foundation for the Study of Diabetes (TW, YF) and the Deutsche Forschungsgemeinschaft (SFB TR23, TP B6 (TW) and A9 (SWS), and GRK 1874 SP2 (TW, YF)).

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animal rights disclosure

Regarding HUVEC isolation, all procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2008 (5). Regarding animal studies, all applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Informed consent disclosure

Informed consent was obtained from all patients for being included in the study.

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    Authors and Affiliations

    1. Institute for Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University of Heidelberg, Mybachstr. 14, 68169, Mannheim, Germany

      Yi Qiu, Di Zhao, Vicki-Marie Butenschön, Thomas Wieland & Yuxi Feng

    2. Division of Experimental Dermatology, Department of Dermatology, Venereology, and Allergology, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany

      Alexander T. Bauer & Stefan W. Schneider

    3. Division of Nephrology, New York University Langone Medical Center, 560 1st Ave, New York, NY, 10016, USA

      Edward Y. Skolnik

    4. 5th Medical Clinic, Medical Faculty Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany

      Hans-Peter Hammes

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    1. Yi Qiu
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    Correspondence to Yuxi Feng.

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    Yi Qiu and Di Zhao have contributed equally to this article.

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    592_2015_752_MOESM1_ESM.jpg

    Supplemental Fig. 1: localization of Ang2 in Weibel-Palade bodies in endothelial cells. Weibel-Palade-bodies were visualized by staining with von Willebrand factor (vWF) (JPEG 103 kb)

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    Qiu, Y., Zhao, D., Butenschön, VM. et al. Nucleoside diphosphate kinase B deficiency causes a diabetes-like vascular pathology via up-regulation of endothelial angiopoietin-2 in the retina. Acta Diabetol 53, 81–89 (2016). https://doi.org/10.1007/s00592-015-0752-x

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