rect targets revealed that RBM20 acts as a Integrated genomic approaches for identification of gene networks ing proteins that contribute to pathological posttranscriptional regulation of an entire network of target genes that lead to cardio- We have identified mutations in RNA bind- myopathy and heart failure. In collabora- deficient in titin splicing and found a loss- tion with Michael Gotthardt (MDC) we used genome-wide approaches in a rat strain of-function mutation in RBM20 as the un- derlying cause for the pathological isoform expression of titin. Mutations in RBM20 in humans have previously been shown to disrupt isoform transitions of a network of genes with essential cardiac functions and cause dilated cardiomyopathy. We showed human disease phenotype. Together with Markus Landthaler and Nikolaus Rajewsky, we investigated the role of RBM20 as a car- that RBM20 deficient rats resembled the diac specific splicing factor. We identified with high-throughput sequencing (CLIP- seq) experiments. The identification of di- splicing repressor in the heart. Using SILAC directly RBM20 regulated transcripts by ap- plying state-of-the-art RNA-protein cross- linking and immunoprecipitation coupled based proteomics we demonstrated direct interaction with important spliceosomal components and showed how mutations within RBM20 may disrupt its repressor function. We further revealed that not only genetic variation, but also the modulation of RBM20 gene expression levels is opera- tive in the regulation of its network and may be an attractive target for therapeutic inter- vention. These binding data will be used to extend our studies determining the impact of genomic variability on posttranscrip- emphasize the importance of posttranscrip- tional regulation in cardiac function and provide novel mechanistic insights into the pathogenesis of human heart failure. tional modifications in humans. Our studies In addition to the identification of thousands pact on histone modifications in rat tissue. sion states. We quantified the genome-wide genes, respectively. In rat tissue, we could show that 14 % (liver) -21 % (heart) of netic influence and segregating within the RI panel. A high proportion of 33 to 49 % of distribution of H3K4me3 and H3K27me3, two methylation marks associated with actively transcribed promoters and silent Several histone marks were associated to functional genomic regions or gene expres- of gene expression traits under genetic con- trol, we recently described the genetic im- Epigenetic control mapped H3K4me3 marks were under ge- MDC Research Report 2016 CARDIOVASCULAR AND METABOLIC DISEASE H3K4me3 marks were under distant genetic control and one major H3K4me3 regulating hundreds of H3K4me3 sites. As the exam- ined histone marks were known predictors of gene expression states, data on genetic variation underlying histone marks could genomic region was identified impacting significantly increase the number of eQTLs. has a great impact on histone modifications in rat tissue that might further help to elu- cidate so far unknown genotype-phenotype relationships in the heart. Hence, we could show that genetic variation Selected Publications Angiogenesis. Nat Commun 7 (2016): 12061. Yusoff, P., Choksi, P. K., Ko, N. S., Singh, M. K., Epstein, D., Guan, Y., Houstek, J., Mracek, T., Nuskova, H., Mikell, B., Tan, J., Pesce, F., Kolar, F., Bottolo, L., Mancini, M., Hubner, Schafer, S., de Marvao, A., Adami, E., Fiedler, L. R., Ng, B., Khin, E., Rackham, O. J., van Heesch, S., Pua, C. J., Kui, M., Walsh, R., Tayal, U., Prasad, S. K., Dawes, T. J., Ko, N. S., Sim, D., Chan, L. L., Chin, C. W., Mazzarotto, F., Barton, P. J., Kreuchwig, F., de Kleijn, D. ner, N., and Cook, S. A., Titin-Truncating Variants Affect Heart Function in Disease Cohorts and the General Population. Nat Genet 49, no. 1 (2017): 46-53. P., Totman, T., Biffi, C., Tee, N., Rueckert, D., Schneider, V., Faber, A., Regitz-Zagrosek, V., Seidman, J. G., Seidman, C. E., Linke, W. A., Kovalik, J. P., O’Regan, D., Ware, J. S., Hub- Wang, M., Sips, P., Khin, E., Rotival, M., Sun, X., Ahmed, R., Widjaja, A. A., Schafer, S., N., Pravenec, M., Petretto, E., MacRae, C., and Cook, S. A., Wars2 Is a Determinant of Hinson, J. T., Chopra, A., Nafissi, N., Polacheck, W. J., Benson, C. C., Swist, S., Gorham, J., Mutations in Ips Cells Define Sarcomere Insufficiency as a Cause of Dilated Cardio- myopathy. Science 349, no. 6251 (2015): 982-6. M., Keogh, A. M., Hayward, C. S., Banner, N. R., Pennell, D. J., O’Regan, D. P., San, T. R., M., Wilson, J. G., O’Donnell, C. J., Prasad, S. K., Barton, P. J., Fatkin, D., Hubner, N., Seidman, J. G., Seidman, C. E., and Cook, S. A., Integrated Allelic, Transcriptional, Roberts, A. M., Ware, J. S., Herman, D. S., Schafer, S., Baksi, J., Bick, A. G., Buchan, R. J., Walsh, R., John, S., Wilkinson, S., Mazzarotto, F., Felkin, L. E., Gong, S., MacArthur, J. A., Cunningham, F., Flannick, J., Gabriel, S. B., Altshuler, D. M., Macdonald, P. S., Heinig, Yang, L., Schafer, S., Sheng, C. C., Haghighi, A., Homsy, J., Hubner, N., Church, G., Cook, S. A., Linke, W. A., Chen, C. S., Seidman, J. G., and Seidman, C. E., Heart Disease. Titin and Phenomic Dissection of the Cardiac Effects of Titin Truncations in Health and Disease. Sci Transl Med 7, no. 270 (2015): 270ra6. Schafer, S., Adami, E., Heinig, M., Rodrigues, K. E., Kreuchwig, F., Silhavy, J., van Heesch, S., Simaite, D., Rajewsky, N., Cuppen, E., Pravenec, M., Vingron, M., Cook, S. A., and Hubner, N., Translational Regulation Shapes the Molecular Landscape of Complex Disease Phenotypes. Nat Commun 6 (2015): 7200. Maatz, H., Jens, M., Liss, M., Schafer, S., Heinig, M., Kirchner, M., Adami, E., Rintisch, C., Dauksaite, V., Radke, M. H., Selbach, M., Barton, P. J., Cook, S. A., Rajewsky, N., Got- thardt, M., Landthaler, M., and Hubner, N., Rna-Binding Protein Rbm20 Represses Splicing to Orchestrate Cardiac Pre-Mrna Processing. J Clin Invest 124, no. 8 (2014): de Marvao, A., Dawes, T. J., Gulati, A., Birks, E. J., Yacoub, M. H., Radke, M., Gotthardt, 3419-30. Group Leader Prof. Dr. Norbert Hübner Franziska Kreuchwig Christin Mieth Valentin Schneider Scientists Allison Faber Katharina Grunz Matthias Heinig Oliver Hummel Henrike Maatz Carola Rintisch Franz Rüschendorf Sebastian Schäfer Sebastiaan van Heesch PhD students Eleonora Adami Maria Bikou Graduate and Undergraduate students Michael Benedikt Mücke Technical Assistants Susanne Blachut Mathias Gerhard Anita Müller Giannino Patone Sabine Schmidt Secretariat Maren Stauch 21