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Liver cell death and anemia in Wilson disease involve acid sphingomyelinase and ceramide

Nature Medicine volume 13, pages 164–170 (2007)Cite this article

Abstract

Wilson disease is caused by accumulation of Cu2+ in cells, which results in liver cirrhosis and, occasionally, anemia. Here, we show that Cu2+ triggers hepatocyte apoptosis through activation of acid sphingomyelinase (Asm) and release of ceramide. Genetic deficiency or pharmacological inhibition of Asm prevented Cu2+-induced hepatocyte apoptosis and protected rats, genetically prone to develop Wilson disease, from acute hepatocyte death, liver failure and early death. Cu2+ induced the secretion of activated Asm from leukocytes, leading to ceramide release in and phosphatidylserine exposure on erythrocytes, events also prevented by inhibition of Asm. Phosphatidylserine exposure resulted in immediate clearance of affected erythrocytes from the blood in mice. Accordingly, individuals with Wilson disease showed elevated plasma levels of Asm, and displayed a constitutive increase of ceramide- and phosphatidylserine-positive erythrocytes. Our data suggest a previously unidentified mechanism for liver cirrhosis and anemia in Wilson disease.

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Figure 1: Cu2+ induces ceramide formation in hepatocytes through activation of Asm.
Figure 2: Cu2+-induced apoptosis of hepatocytes requires Asm.
Figure 3: Cu2+ induces ceramide formation in human erythrocytes.
Figure 4: Cu2+ triggers anemia via Asm.
Figure 5: Asm activity, ceramide and phosphatidylserine exposure in individuals with Wilson disease.
Figure 6: Inhibition of Asm delays Wilson disease in genetically prone rats.

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Acknowledgements

The authors acknowledge the technical assistance of B. Wilker, S. Keitsch, E. Faber and W. Gfrörer and preparation of the manuscript by T. Loch and S. Moyrer. Goat antibodies to Asm were provided by K. Sandhoff (University of Bonn). This study was supported by the Deutsche Forschungsgemeinschaft (DFG) grants La 315/4-3, La 315/6-1 (to F.L.) and La 315/13-1 (to F.L. and T.W.), the Bundesministerium für Bildung, Wissenschaft, Forschung und Technologie (Center for Interdisciplinary Clinical Research) 01 KS 9602 and IZKF-Promotionskolleg “Molekulare Medizin # 1547” (to F.L., T.W., P.A.L. and D.S.K.), Wilhelm Sander-Stiftung (to T.W.), an Interne Forschungsförderung Essen grant (to M.S.) and DFG grant GU 335/10-3 (to E.G.).

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Author notes

  1. Thomas Wieder

    Present address: Present address: Department of Dermatology, University of Tübingen, 72076 Tübingen, Germany.,

  2. Erich Gulbins and Florian Lang: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Physiology University of Tübingen, Tübingen, 72076, Germany

    Philipp A Lang, Jan P Nicolay, Daniela S Kempe, Adrian Lupescu, Saisudha Koka, Kerstin Eisele, Barbara A Klarl, Stephan M Huber, Thomas Wieder & Florian Lang

  2. Institute of Molecular Biology, University of Duisburg-Essen, Essen, 45122, Germany

    Marcus Schenck & Erich Gulbins

  3. Department of Urology, University of Duisburg-Essen, Essen, 45122, Germany

    Marcus Schenck & Herbert Rübben

  4. Institute of Pathology and Neuropathology, University of Duisburg-Essen, Essen, 45122, Germany

    Jan Ulrich Becker & Kurt W Schmid

  5. Department of Internal Medicine, University Clinic, University of Duisburg-Essen, Essen, 45122, Germany

    Klaus Mann

  6. Department of Occupational and Social Medicine, University of Tübingen, Tübingen, 72076, Germany

    Sibylle Hildenbrand

  7. Department of Neurology, University of Düsseldorf, Düsseldorf, 40225, Germany

    Harald Hefter

  8. Department of Gastroenterology, Hepatology and Infectiology, University of Düsseldorf, Düsseldorf, 40225, Germany

    Andreas Erhardt & Dieter Häussinger

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Contributions

P.A.L., F.L. and E.G. initiated the studies. P.A.L. investigated the effects of copper on Annexin V binding to human erythrocytes and HepG2 cells. M.S. performed the rat experiments and analyzed the results. J.P.N. analyzed Annexin V binding to patient erythrocytes. J.U.B. and K.W.S. did the histology studies. D.S.K. performed ceramide measurents in HepG2 cells. A.L. performed Ca2+ measurements in HepG2 cells. S.K. and S.M.H. studied in vivo clearance of circulating erythrocytes in rats. K.E. and B.A.K. performed ceramide measurements in human erythrocytes. H.R. supervised rat experiments. K.M. measured bilirubin and amitriptyline in rat plasma. S.H. made the copper determinations. H.H., A.E. and D.H. provided blood and data from individuals with Wilson disease. T.W. supervised part of the laboratory work in Tübingen and performed statistical analysis. E.G. measured ASM activity and ceramide, did the mice experiments, analyzed the data and supervised the group in Essen. F.L. supervised the group in Tübingen. F.L. and E.G. designed the study and wrote the manuscript.

Corresponding authors

Correspondence to Erich Gulbins or Florian Lang.

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The authors declare no competing financial interests.

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Lang, P., Schenck, M., Nicolay, J. et al. Liver cell death and anemia in Wilson disease involve acid sphingomyelinase and ceramide. Nat Med 13, 164–170 (2007). https://doi.org/10.1038/nm1539

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