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Bidirectional signalling between EphA2 and ephrinA1 increases tubular cell attachment, laminin secretion and modulates erythropoietin expression after renal hypoxic injury

  • Molecular and cellular mechanisms of disease
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Pflügers Archiv - European Journal of Physiology Aims and scope Submit manuscript

Abstract

Acute kidney injury (AKI) is common in hospitalized patients and has a poor prognosis, the severity of AKI being linked to progression to chronic kidney disease. This stresses the need to search for protective mechanisms during the acute phase. We investigated kidney repair after hypoxic injury using a rat model of renal artery branch ligation, which led to an oxygen gradient vertical to the corticomedullary axis. Three distinct zones were observed: tubular necrosis, infarction border zone and preserved normal tissue. EphA2 is a receptor tyrosine kinase with pivotal roles in cell architecture, migration and survival, upon juxtacrine contact with its membrane-bound ligand EphrinA1. Following hypoxia, EphA2 was up-regulated in cortical and medullary tubular cells, while EphrinA1 was up-regulated in interstitial cells adjacent to peritubular capillaries. Moreover, erythropoietin (EPO) messenger RNA (mRNA) was strongly expressed in the border zone of infarcted kidney within the first 6 h. To gain more insight into the biological impact of EphA2 and EphrinA1 up-regulation, we activated the signalling pathways in vitro using recombinant EphrinA1/Fc or EphA2/Fc proteins. Stimulation of EphA2 forward signalling in the proximal tubular cell line HK2 increased cell attachment and laminin secretion at the baso-lateral side. Conversely, activation of reverse signalling through EphrinA1 expressed by Hep3B cells promoted EPO production at both the transcriptional and protein level. Strikingly, in co-culture experiments, juxtacrine contact between EphA2 expressing MDCK and EphrinA1 expressing Hep3B was sufficient to induce a significant up-regulation of EPO mRNA production in the latter cells, even in the absence of hypoxic conditions. The synergistic effects of EphA2 and hypoxia led to a 15–20-fold increase of EPO expression. Collectively, our results suggest an important role of EphA2/EphrinA1 signalling in kidney repair after hypoxic injury through stimulation of (i) tubular cell attachment, (ii) secretion of basal membrane proteins and (iii) EPO production. These findings could thus pave the way to new therapeutic approaches.

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Acknowledgments

We would like to thank T. Schiegg for her enthusiastic help in the in vitro experiments. We thank C. Rosenberger (Center for Cardiovascular Research, Charité Berlin) for his precious advices during the set-up of the animal experiments, and B.M. Frey and F.J. Frey (Division of Nephrology and Hypertension, Inselspital, University of Bern, Switzerland) for their support and fruitful inputs. This project has been supported by Grant 31003A-105871 of the Swiss National Science Foundation to UHD, and Grant 34-482 of the Novartis Foundation for Biomedical Research to KF and UHD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors declare no financial conflicts of interest.

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    Authors and Affiliations

    1. Division of Nephrology, Hypertension and Clinical Pharmacology, Inselspital, University of Bern Medical School, CH-3010, Bern, Switzerland

      Stéphane Rodriguez, Stefan Rudloff, Katrin Franziska Koenig, Swapna Karthik & Uyen Huynh-Do

    2. Institute of Physiology, University of Zurich, Zurich, Switzerland

      David Hoogewijs

    3. Present address: U917 Inserm unit, University of Rennes, Rennes, France

      Stéphane Rodriguez

    4. Present address: Division of Nephrology, University Hospital of Basel, Basel, Switzerland

      Katrin Franziska Koenig

    5. Present address: Institute of Physiology, University of Duisburg-Essen, Essen, Germany

      David Hoogewijs

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    1. Stéphane Rodriguez
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    Stéphane Rodriguez and Stefan Rudloff contributed equally to this work.

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    Rodriguez, S., Rudloff, S., Koenig, K.F. et al. Bidirectional signalling between EphA2 and ephrinA1 increases tubular cell attachment, laminin secretion and modulates erythropoietin expression after renal hypoxic injury. Pflugers Arch - Eur J Physiol 468, 1433–1448 (2016). https://doi.org/10.1007/s00424-016-1838-1

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