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eNOS-induced vascular barrier disruption in retinopathy by c-Src activation and tyrosine phosphorylation of VE-cadherin

  1. Takeshi Ninchoji  Is a corresponding author
  2. Dominic T Love
  3. Ross O Smith
  4. Marie Hedlund
  5. Dietmar Vestweber
  6. William C Sessa
  7. Lena Claesson-Welsh  Is a corresponding author
  1. Uppsala University, Sweden
  2. Max Planck Institute for Molecular Biomedicine, Germany
  3. Yale School of Medicine, United States
Research Article
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Cite this article as: eLife 2021;10:e64944 doi: 10.7554/eLife.64944

Abstract

Background: Hypoxia and consequent production of vascular endothelial growth factor A (VEGFA) promote blood vessel leakiness and edema in ocular diseases. Anti-VEGFA therapeutics may aggravate hypoxia; therefore, therapy development is needed.

Methods: Oxygen-induced retinopathy was used as a model to test the role of nitric oxide (NO) in pathological neovascularization and vessel permeability. Suppression of NO formation was achieved chemically using L-NMMA, or genetically, in endothelial NO synthase (eNOS) serine to alanine (S1176A) mutant mice.

Results: Suppression of NO formation resulted in reduced retinal neoangiogenesis. Remaining vascular tufts exhibited reduced vascular leakage through stabilized endothelial adherens junctions, manifested as reduced phosphorylation of vascular endothelial (VE)-cadherin Y685 in a c-Src-dependent manner. Treatment with a single dose of L-NMMA in established retinopathy restored the vascular barrier and prevented leakage.

Conclusion: We conclude that NO destabilizes adherens junctions, resulting in vascular hyperpermeability, by converging with the VEGFA/VEGFR2/c-Src/VE-cadherin pathway.

Funding: This study was supported by the Swedish Cancer foundation (19 0119 Pj 01 H), the Swedish Research Council (2020-01349), the Knut and Alice Wallenberg foundation (KAW 2020.0057) and a Fondation Leducq Transatlantic Network of Excellence Grant in Neurovascular Disease (17 CVD 03). KAW also supported LCW with a Wallenberg Scholar grant (2015.0275). WCS was supported by Grants R35 HL139945, P01 HL1070205, AHA MERIT Award.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 1-5. Source data files have also been deposited with Dryad: https://doi.org/10.5061/dryad.x69p8czhv

The following data sets were generated

Article and author information

Author details

  1. Takeshi Ninchoji

    Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    For correspondence
    nincho830@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
  2. Dominic T Love

    Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  3. Ross O Smith

    Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4239-3204
  4. Marie Hedlund

    Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  5. Dietmar Vestweber

    Department of Vascular Cell Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3517-732X
  6. William C Sessa

    Pharmacology, Yale School of Medicine, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5759-1938
  7. Lena Claesson-Welsh

    Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    For correspondence
    lena.welsh@igp.uu.se
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4275-2000

Funding

Swedish Cancer Foundation (19 0119 Pj 01 H)

  • Lena Claesson-Welsh

Vetenskapsrådet (2020-01349)

  • Lena Claesson-Welsh

Knut och Alice Wallenbergs Stiftelse (KAW 2015.0030)

  • Lena Claesson-Welsh

Knut och Alice Wallenbergs Stiftelse (KAW.0275)

  • Lena Claesson-Welsh

Fondation Leducq (17 CVD 03)

  • Lena Claesson-Welsh

National Institutes of Health (R35 HL 139945)

  • William C Sessa

National Institutes of Health (P01HL 1070205)

  • William C Sessa

American Heart Association (Merit Award)

  • William C Sessa

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: Mouse husbandry and oxygen-induced retinopathy (OIR) challenge took place at Uppsala University,and the University board of animal experimentation approved all animal work for the studies (ethical permit 5.2.18-8927/16). Animal handling was in accordance to the ARVO statement for the Use of Animals in Ophthalmologic and Vision Research. Professional animal care was provided and overseen by University veterinarians. Every effort was made to minimize suffering of the animal.

Reviewing Editor

  1. Gou Young Koh, Institute of Basic Science and Korea Advanced Institute of Science and Technology (KAIST), Korea (South), Republic of

Publication history

  1. Received: November 16, 2020
  2. Accepted: April 22, 2021
  3. Accepted Manuscript published: April 28, 2021 (version 1)
  4. Version of Record published: April 30, 2021 (version 2)

Copyright

© 2021, Ninchoji et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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