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RBM20, a gene for hereditary cardiomyopathy, regulates titin splicing

Nature Medicine volume 18, pages 766–773 (2012)Cite this article

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Abstract

Alternative splicing has a major role in cardiac adaptive responses, as exemplified by the isoform switch of the sarcomeric protein titin, which adjusts ventricular filling. By positional cloning using a previously characterized rat strain with altered titin mRNA splicing, we identified a loss-of-function mutation in the gene encoding RNA binding motif protein 20 (Rbm20) as the underlying cause of pathological titin isoform expression. The phenotype of Rbm20-deficient rats resembled the pathology seen in individuals with dilated cardiomyopathy caused by RBM20 mutations. Deep sequencing of the human and rat cardiac transcriptome revealed an RBM20-dependent regulation of alternative splicing. In addition to titin (TTN), we identified a set of 30 genes with conserved splicing regulation between humans and rats. This network is enriched for genes that have previously been linked to cardiomyopathy, ion homeostasis and sarcomere biology. Our studies emphasize the key role of post-transcriptional regulation in cardiac function and provide mechanistic insights into the pathogenesis of human heart failure.

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Figure 1: Mapping of the titin splice defect and validation of RBM20 as the affected gene.
Figure 2: Analysis of Rbm20 expression and function.
Figure 3: Signs of cardiomyopathy with arrhythmia and sudden death in Rbm20-deficient rats.
Figure 4: Identification of a novel and functionally relevant RBM20 mutation in an individual with severe cardiomyopathy and arrhythmia.
Figure 5: Characterization of RBM20-dependent isoform expression.
Figure 6: Alignment of orthologous rat and human exons to compare RBM20-dependent isoform expression.

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Acknowledgements

We are grateful to B. Goldbrich, O. Hummel, S. Lubitz, S. Makino, G. Patone, S. Blachut, R. Plehm, S. Probst, S. Schmidt and M. Wehle for expert technical assistance. R. Hetzer (Deutsches Herzzentrum Berlin) generously provided human cardiac tissue. Mammalian expression vectors for PTBP1 and HuD were gifts from R. Darnell, The Rockefeller University. This work was supported by the US National Institutes of Health grants HL77196 (M.L.G.) and HL075431 (C.A.M.), the Deutsche Forschungsgemeinschaft, Bonn, Germany and the European Research Council grant StG282078 (M.G.), the German Ministry of Science and Education (Nationales Genomforschungsnetz, NGFN-Plus Heart Failure Network) and EURATRANS (HEALTH-F4-2010-241504) (N.H.) and the American Heart Association (P.T.E.).

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Author notes

  1. Wei Guo and Sebastian Schafer: These authors contributed equally to this work.

Authors and Affiliations

  1. Muscle Biology Laboratory, University of Wisconsin Madison, Madison, Wisconsin, USA

    Wei Guo, Marion L Greaser, Shijun Li & Amanda M Parrish

  2. Cardiovascular and Metabolic Sciences, Max Delbrück Center for Molecular Medicine, Berlin, Germany

    Sebastian Schafer, Henrike Maatz, Herbert Schulz, Kathrin Saar & Norbert Hubner

  3. Neuromuscular and Cardiovascular Cell Biology, Max Delbrück Center for Molecular Medicine, Berlin, Germany

    Michael H Radke, Martin Liss, Thirupugal Govindarajan, Vita Dauksaite, Padmanabhan Vakeel & Michael Gotthardt

  4. Cardiovascular Molecular Genetics, Max Delbrück Center for Molecular Medicine, Berlin, Germany

    Sabine Klaassen, Brenda Gerull & Ludwig Thierfelder

  5. Institute of Gender in Medicine and Center for Cardiovascular Research, Charité, University Medicine Berlin, Berlin, Germany

    Vera Regitz-Zagrosek

  6. Department of Medicine, University of Wisconsin Madison, Madison, Wisconsin, USA

    Timothy A Hacker & Kurt W Saupe

  7. Cardiology Division, Massachusetts General Hospital, Charlestown, Massachusetts, USA

    G William Dec, Patrick T Ellinor & Calum A MacRae

  8. Universitätskilikum Benjamin Franklin, Charité, University Medicine Berlin, Berlin, Germany

    Bastian Spallek & Robert Fischer

  9. Department of Cardiology (Campus Virchow Klinikum), Charité University Medicine Berlin, Berlin, Germany

    Andreas Perrot & Cemil Özcelik

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  1. Wei Guo
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Contributions

M.L.G., N.H. and M.G. designed the research. W.G., S.S., M.H.R., M.L., T.G., H.M., H.S., S.L., A.M.P., V.D., P.V., S.K., B.G., L.T., V.R.-Z., T.A.H., K.W.S., G.W.D., P.T.E., C.A.M., B.S., R.F., A.P., C.O. and K.S. performed the research. W.G., S.S., M.L.G., H.S., M.H.R., M.L., T.G., H.M., S.L., A.M.P., V.D., P.V., S.K., B.G., L.T., V.R.-Z., T.A.H., K.W.S., G.W.D., P.T.E., C.A.M., B.S., R.F., A.P., C.O., K.S. and M.G. performed data analysis. W.G., T.G., H.M., H.S., S.L., A.M.P., V.D., P.V., S.K., B.G., L.T., V.R.-Z., T.A.H., K.W.S., G.W.D., P.T.E., C.A.M., B.S., R.F., A.P., C.O. and K.S. provided discussion and advice. V.R.-Z., A.P. and C.O. provided patient material. S.S., M.H.R., T.G., H.S. and M.G. performed the bioinformatics analysis. S.S., M.L.G., M.H.R., N.H. and M.G. wrote the paper.

Corresponding author

Correspondence to Michael Gotthardt.

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Guo, W., Schafer, S., Greaser, M. et al. RBM20, a gene for hereditary cardiomyopathy, regulates titin splicing. Nat Med 18, 766–773 (2012). https://doi.org/10.1038/nm.2693

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