Marlies Sachs1, Sebastian Wetzel2, Julia Reichelt1, Wiebke Sachs1, Lisa Schebsdat1, Stephanie Zielinski1, Lisa Seipold2, Lukas Heintz1, Stephan A Müller3,4, Oliver Kretz5, Maja Lindenmeyer5, Thorsten Wiech6, Tobias B Huber5, Renate Lüllmann-Rauch7, Stefan F Lichtenthaler3,4,8, Paul Saftig9, Catherine Meyer-Schwesinger10. 1. Institute of Cellular and Integrative Physiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 2. Institute of Biochemistry, Christian-Albrechts University Kiel, Kiel, Germany. 3. German Center for Neurodegenerative Diseases (DZNE), Munich, Germany. 4. Neuroproteomics, School of Medicine, University Hospital rechts der Isar, Technical University of Munich, Munich, Germany. 5. III. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 6. Nephropathology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany. 7. Institute of Anatomy, Christian-Albrechts University Kiel, Kiel, Germany. 8. Munich Cluster for Systems Neurology (SyNergy), Munich, Germany. 9. Institute of Biochemistry, Christian-Albrechts University Kiel, Kiel, Germany psaftig@biochem.uni-kiel.de c.meyer-schwesinger@uke.uni-hamburg.de. 10. Institute of Cellular and Integrative Physiology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany psaftig@biochem.uni-kiel.de c.meyer-schwesinger@uke.uni-hamburg.de.
Abstract
BACKGROUND: Podocytes embrace the glomerular capillaries with foot processes, which are interconnected by a specialized adherens junction to ultimately form the filtration barrier. Altered adhesion and loss are common features of podocyte injury, which could be mediated by shedding of cell-adhesion molecules through the regulated activity of cell surface-expressed proteases. A Disintegrin and Metalloproteinase 10 (ADAM10) is such a protease known to mediate ectodomain shedding of adhesion molecules, among others. Here we evaluate the involvement of ADAM10 in the process of antibody-induced podocyte injury. METHODS: Membrane proteomics, immunoblotting, high-resolution microscopy, and immunogold electron microscopy were used to analyze human and murine podocyte ADAM10 expression in health and kidney injury. The functionality of ADAM10 ectodomain shedding for podocyte development and injury was analyzed, in vitro and in vivo, in the anti-podocyte nephritis (APN) model in podocyte-specific, ADAM10-deficient mice. RESULTS: ADAM10 is selectively localized at foot processes of murine podocytes and its expression is dispensable for podocyte development. Podocyte ADAM10 expression is induced in the setting of antibody-mediated injury in humans and mice. Podocyte ADAM10 deficiency attenuates the clinical course of APN and preserves the morphologic integrity of podocytes, despite subepithelial immune-deposit formation. Functionally, ADAM10-related ectodomain shedding results in cleavage of the cell-adhesion proteins N- and P-cadherin, thus decreasing their injury-related surface levels. This favors podocyte loss and the activation of downstream signaling events through the Wnt signaling pathway in an ADAM10-dependent manner. CONCLUSIONS: ADAM10-mediated ectodomain shedding of injury-related cadherins drives podocyte injury.
BACKGROUND: Podocytes embrace the glomerular capillaries with foot processes, which are interconnected by a specialized adherens junction to ultimately form the filtration barrier. Altered adhesion and loss are common features of podocyte injury, which could be mediated by shedding of cell-adhesion molecules through the regulated activity of cell surface-expressed proteases. A Disintegrin and Metalloproteinase 10 (ADAM10) is such a protease known to mediate ectodomain shedding of adhesion molecules, among others. Here we evaluate the involvement of ADAM10 in the process of antibody-induced podocyte injury. METHODS: Membrane proteomics, immunoblotting, high-resolution microscopy, and immunogold electron microscopy were used to analyze human and murine podocyte ADAM10 expression in health and kidney injury. The functionality of ADAM10 ectodomain shedding for podocyte development and injury was analyzed, in vitro and in vivo, in the anti-podocyte nephritis (APN) model in podocyte-specific, ADAM10-deficient mice. RESULTS: ADAM10 is selectively localized at foot processes of murine podocytes and its expression is dispensable for podocyte development. Podocyte ADAM10 expression is induced in the setting of antibody-mediated injury in humans and mice. Podocyte ADAM10 deficiency attenuates the clinical course of APN and preserves the morphologic integrity of podocytes, despite subepithelial immune-deposit formation. Functionally, ADAM10-related ectodomain shedding results in cleavage of the cell-adhesion proteins N- and P-cadherin, thus decreasing their injury-related surface levels. This favors podocyte loss and the activation of downstream signaling events through the Wnt signaling pathway in an ADAM10-dependent manner. CONCLUSIONS: ADAM10-mediated ectodomain shedding of injury-related cadherins drives podocyte injury.
Authors: Nicola M Tomas; Laurence H Beck; Catherine Meyer-Schwesinger; Barbara Seitz-Polski; Hong Ma; Gunther Zahner; Guillaume Dolla; Elion Hoxha; Udo Helmchen; Anne-Sophie Dabert-Gay; Delphine Debayle; Michael Merchant; Jon Klein; David J Salant; Rolf A K Stahl; Gérard Lambeau Journal: N Engl J Med Date: 2014-11-13 Impact factor: 91.245
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Authors: Laurence H Beck; Ramon G B Bonegio; Gérard Lambeau; David M Beck; David W Powell; Timothy D Cummins; Jon B Klein; David J Salant Journal: N Engl J Med Date: 2009-07-02 Impact factor: 91.245
Authors: Juan Antonio Vizcaíno; Attila Csordas; Noemi del-Toro; José A Dianes; Johannes Griss; Ilias Lavidas; Gerhard Mayer; Yasset Perez-Riverol; Florian Reisinger; Tobias Ternent; Qing-Wei Xu; Rui Wang; Henning Hermjakob Journal: Nucleic Acids Res Date: 2015-11-02 Impact factor: 16.971