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Nexilin mutations destabilize cardiac Z-disks and lead to dilated cardiomyopathy

Nature Medicine volume 15, pages 1281–1288 (2009)Cite this article

Abstract

Z-disks, the mechanical integration sites of heart and skeletal muscle cells, link anchorage of myofilaments to force reception and processing. The key molecules that enable the Z-disk to persistently withstand the extreme mechanical forces during muscle contraction have not yet been identified. Here we isolated nexilin (encoded by NEXN) as a novel Z-disk protein. Loss of nexilin in zebrafish led to perturbed Z-disk stability and heart failure. To evaluate the role of nexilin in human heart failure, we performed a genetic association study on individuals with dilated cardiomyopathy and found several mutations in NEXN associated with the disease. Nexilin mutation carriers showed the same cardiac Z-disk pathology as observed in nexilin-deficient zebrafish. Expression in zebrafish of nexilin proteins encoded by NEXN mutant alleles induced Z-disk damage and heart failure, demonstrating a dominant-negative effect and confirming the disease-causing nature of these mutations. Increasing mechanical strain aggravated Z-disk damage in nexilin-deficient skeletal muscle, implying a unique role of nexilin in protecting Z-disks from mechanical trauma.

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Figure 1: Nexilin is found at heart and skeletal muscle Z-disks.
Figure 2: Inactivation of nexn leads to dilated cardiomyopathy in zebrafish.
Figure 3: NEXN mutations are associated with dilated cardiomyopathy in humans.
Figure 4: Mutant NEXN has a dominant-negative effect on zebrafish hearts.
Figure 5: Nexilin has a unique role in stabilizing cardiac Z-disks.

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Acknowledgements

We thank S. Marquart, S. Weber, S. Kolb, H. Hosser, O. Mücke and S. Manthey as well as S. Wrobel for their excellent technical support. We thank B. Jurmann for human subject work-up and evaluation of clinical data and N. Frey (University of Heidelberg) for providing the calsarcin-specific antibody. We thank R. Knöll and L. Thierfelder for critical discussion of patient data. This work was supported by Deutsche Forschungsgemeinschaft Ro2173/1-1, Ro2173/2-1, Ro2173/2-2 and Ro2173/3-1 (W.R.), Bundesministerium für Bildung und Forschung 01GS0108, 01GS0420 and 01GS0836 (W.R., B.M., D.W. and H.A.K.) and the Postdoc-Fellowship YIA of the medical faculty of the University of Heidelberg (T.D.). We are indebted to all probands for their participation. We thank M.C. Fishman for critical discussion of the manuscript.

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Author notes

  1. David Hassel, Tillman Dahme and Jeanette Erdmann: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine III, University of Heidelberg, Heidelberg, Germany

    David Hassel, Tillman Dahme, Benjamin Meder, Steffen Just, Alexander Hess, Philipp Ehlermann, Dieter Weichenhan, Hugo A Katus & Wolfgang Rottbauer

  2. Medizinische Klinik II, Universität zu Lübeck, Germany

    Jeanette Erdmann, Henrike Liptau & Heribert Schunkert

  3. Department of Genetic Epidemiology of Vascular Diseases, Leibniz-Institute for Arteriosclerosis Research at the University of Münster, Münster, Germany

    Andreas Huge & Monika Stoll

  4. Department of Biochemistry, Biocenter at the University of Würzburg, Würzburg, Germany

    Matthias Grimmler

  5. German Heart Center Berlin, Berlin, Germany

    Roland Hetzer & Vera Regitz-Zagrosek

  6. Center for Cardiovascular Research, Charité, Berlin, Germany

    Vera Regitz-Zagrosek

  7. Department of Human Genetics, University of Heidelberg, Heidelberg, Germany

    Christine Fischer

  8. Cologne Center for Genomics, University of Cologne, Cologne, Germany

    Peter Nürnberg

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  1. David Hassel
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Contributions

D.H. started the project and performed the bioinformatic screen, zebrafish and human expression analysis, subcellular localization by immunofluorescence and transmission electron microscopy, zebrafish knockdown experiments, human microsatellite marker analysis, overexpression of human mutations in zebrafish and zebrafish skeletal muscle pacing experiments. T.D. conducted the zebrafish nexn knockdown characterization, subcellular localization by immunofluorescence and transmission electron microscopy, human expression analysis, human microsatellite marker analysis, overexpression of human mutations in zebrafish and zebrafish skeletal muscle pacing experiments and wrote the manuscript. J.E. initiated and performed the human genetics association study and the microsatellite marker analysis in NEXN-G650del mutation carriers. B.M. performed the human genetics association study. A. Huge performed the statistical analysis of the whole-genome SNP screen by microarray. M.S. conducted and interpreted the statistical analysis of the whole-genome microarray. S.J. contributed to the zebrafish experiments. A. Hess and P.E. analyzed the family structures of NEXN mutation carriers. D.W. performed the human genetics association study. M.G. conducted the immunoprecpitation experiments. H.L. contributed to the human genetics association study. R.H., V.R.-Z., H.S. and H.A.K. contributed subjects to the human genetics study. C.F. analyzed the human genetics data. P.N. conducted the whole-genome microarray screen and contributed the control group for the microarray screen. W.R. supervised the project, analyzed the data and wrote the manuscript.

Corresponding author

Correspondence to Wolfgang Rottbauer.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–4 (PDF 709 kb)

Supplementary Movie 1

Loss of nexn leads to DCM in zebrafish. The hearts of MO-nexn and MO-ctrl injected zebrafish embryos at 48 hpf and 72 hpf are shown. The cardiac ventricles (v) and atria (a) are indicated. (AVI 24694 kb)

Supplementary Movie 2

Expression of the human NEXN mutations in zebrafish demonstrate a dominant-negative mode of action. (AVI 13936 kb)

Injection of wild-type NEXN (NEXN wt) has no effect and zebrafish cardiac function. In contrast, injection of equal amounts of NEXN-G650del, NEXN-Y652C, or NEXN-P611T leads to DCM in zebrafish. The cardiac ventricles (v) and atria (a) are indicated.

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Hassel, D., Dahme, T., Erdmann, J. et al. Nexilin mutations destabilize cardiac Z-disks and lead to dilated cardiomyopathy. Nat Med 15, 1281–1288 (2009). https://doi.org/10.1038/nm.2037

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