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Akt1 promotes stimuli-induced endothelial-barrier protection through FoxO-mediated tight-junction protein turnover

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Abstract

Vascular permeability regulated by the vascular endothelial growth factor (VEGF) through endothelial-barrier junctions is essential for inflammation. Mechanisms regulating vascular permeability remain elusive. Although ‘Akt’ and ‘Src’ have been implicated in the endothelial-barrier regulation, it is puzzling how both agents that protect and disrupt the endothelial-barrier activate these kinases to reciprocally regulate vascular permeability. To delineate the role of Akt1 in endothelial-barrier regulation, we created endothelial-specific, tamoxifen-inducible Akt1 knockout mice and stable ShRNA-mediated Akt1 knockdown in human microvascular endothelial cells. Akt1 loss leads to decreased basal and angiopoietin1-induced endothelial-barrier resistance, and enhanced VEGF-induced endothelial-barrier breakdown. Endothelial Akt1 deficiency resulted in enhanced VEGF-induced vascular leakage in mice ears, which was rescued upon re-expression with Adeno-myrAkt1. Furthermore, co-treatment with angiopoietin1 reversed VEGF-induced vascular leakage in an Akt1-dependent manner. Mechanistically, our study revealed that while VEGF-induced short-term vascular permeability is independent of Akt1, its recovery is reliant on Akt1 and FoxO-mediated claudin expression. Pharmacological inhibition of FoxO transcription factors rescued the defective endothelial barrier due to Akt1 deficiency. Here we provide novel insights on the endothelial-barrier protective role of VEGF in the long term and the importance of Akt1-FoxO signaling on tight-junction stabilization and prevention of vascular leakage through claudin expression.

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Abbreviations

VEGF:

Vascular endothelial growth factor

Ang-1:

Angiopoietin-1

VECad-Cre-Ak1:

Vascular endothelial cadherin-cre recombinase-Akt1 knockdown

HMEC:

Human microvascular endothelial cells

FoxO:

Forkhead box protein O

ECIS:

Electric cell-substrate impedance sensing

WT:

Wild type

GFP:

Green fluorescent protein

eNOS:

Endothelial nitric oxide synthase

AJ:

Adherens junction

TJ:

Tight junction

Zo:

Zona occludens

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Acknowledgments

Funds were provided by the National Institutes of Health Grant (R01HL103952) to PRS. This material is the result of work supported with resources and the use of facilities at the Charlie Norwood VAMC, Augusta, GA. The funders had no role in the study design, data collection, analysis and decision to publish. Preparation of the manuscript and the contents do not represent the views of the Department of Veterans Affairs or the United States Government.

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Authors and Affiliations

  1. Clinical and Experimental Therapeutics, College of Pharmacy, University of Georgia and Charlie Norwood VA Medical Center, Augusta, GA, USA

    Fei Gao, Sandeep Artham, Harika Sabbineni, Ahmad Al-Azayzih & Payaningal R. Somanath

  2. Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Fei Gao

  3. College of Pharmacy, Jordan University of Science and Technology, Irbid, Jordan

    Ahmad Al-Azayzih

  4. Department of Biochemistry and Molecular Genetics, College of Medicine, University of Illinois at Chicago, Chicago, IL, USA

    Xiao-Ding Peng & Nissim Hay

  5. Max Plank Institute of Molecular Biomedicine, Röntgenstraße 20, Münster, Germany

    Ralf H. Adams

  6. Department of Molecular Cardiology, Joseph J. Jacob’s Center for Thrombosis and Vascular Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA

    Tatiana V. Byzova

  7. Department of Medicine and Vascular Biology Center, Augusta University, HM1200, Augusta, GA, 30912, USA

    Payaningal R. Somanath

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  1. Fei Gao
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Correspondence to Payaningal R. Somanath.

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Gao, F., Artham, S., Sabbineni, H. et al. Akt1 promotes stimuli-induced endothelial-barrier protection through FoxO-mediated tight-junction protein turnover. Cell. Mol. Life Sci. 73, 3917–3933 (2016). https://doi.org/10.1007/s00018-016-2232-z

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  • DOI: https://doi.org/10.1007/s00018-016-2232-z

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