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Severe DCM phenotype of patient harboring RBM20 mutation S635A can be modeled by patient-specific induced pluripotent stem cell-derived cardiomyocytes.
J Mol Cell Cardiol. 2017 12; 113:9-21.JM

Abstract

The ability to generate patient-specific induced pluripotent stem cells (iPSCs) provides a unique opportunity for modeling heart disease in vitro. In this study, we generated iPSCs from a patient with dilated cardiomyopathy (DCM) caused by a missense mutation S635A in RNA-binding motif protein 20 (RBM20) and investigated the functionality and cell biology of cardiomyocytes (CMs) derived from patient-specific iPSCs (RBM20-iPSCs). The RBM20-iPSC-CMs showed abnormal distribution of sarcomeric α-actinin and defective calcium handling compared to control-iPSC-CMs, suggesting disorganized myofilament structure and altered calcium machinery in CMs of the RBM20 patient. Engineered heart muscles (EHMs) from RBM20-iPSC-CMs showed that not only active force generation was impaired in RBM20-EHMs but also passive stress of the tissue was decreased, suggesting a higher visco-elasticity of RBM20-EHMs. Furthermore, we observed a reduced titin (TTN) N2B-isoform expression in RBM20-iPSC-CMs by demonstrating a reduction of exon skipping in the PEVK region of TTN and an inhibition of TTN isoform switch. In contrast, in control-iPSC-CMs both TTN isoforms N2B and N2BA were expressed, indicating that the TTN isoform switch occurs already during early cardiogenesis. Using next generation RNA sequencing, we mapped transcriptome and splicing target profiles of RBM20-iPSC-CMs and identified different cardiac gene networks in response to the analyzed RBM20 mutation in cardiac-specific processes. These findings shed the first light on molecular mechanisms of RBM20-dependent pathological cardiac remodeling leading to DCM. Our data demonstrate that iPSC-CMs coupled with EHMs provide a powerful tool for evaluating disease-relevant functional defects and for a deeper mechanistic understanding of alternative splicing-related cardiac diseases.

Authors+Show Affiliations

Department of Cardiology and Pneumology, Universitätsmedizin Göttingen, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany.DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany; Institute of Pharmacology and Toxicology, Universitätsmedizin Göttingen, Germany.DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany; Department of Cardiovascular Physiology, Ruhr University Bochum, Germany.Institute of Pharmacology and Toxicology, Technische Universität Dresden, Germany.Institute of Pharmacology and Toxicology, Technische Universität Dresden, Germany.Department of Cardiology and Pneumology, Universitätsmedizin Göttingen, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany.Cardiovascular Genetics, Experimental and Clinical Research Center, Charité-Universitätsmedizin Berlin, Germany; Medizinischen Klinik I Kardiologie, Gastroenterologie und Diabetologie, Knappschaftskrankenhaus Recklingshausen, Germany.Cardiovascular Genetics, Experimental and Clinical Research Center, Charité-Universitätsmedizin Berlin, Germany.Department of Cardiology and Pneumology, Universitätsmedizin Göttingen, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany; Department of Internal Medicine 2 - Cardiology, University Medical Center Regensburg, Germany.Department of Cardiology, University of Heidelberg, Germany; DZHK, Partner Site Heidelberg, Germany.German Center for Neurodegenerative Diseases, Göttingen and Tübingen, Germany.Department of Cardiology and Pneumology, Universitätsmedizin Göttingen, Germany.Department of Cardiology and Pneumology, Universitätsmedizin Göttingen, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany.Department of Cardiology, University of Heidelberg, Germany; DZHK, Partner Site Heidelberg, Germany.German Center for Neurodegenerative Diseases, Göttingen and Tübingen, Germany; Institute of Medical Systems Biology, Center for Molecular Neurobiology, University Clinic Hamburg-Eppendorf, Hamburg, Germany.DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany; Department of Cardiovascular Physiology, Ruhr University Bochum, Germany.DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany; Institute of Pharmacology and Toxicology, Universitätsmedizin Göttingen, Germany.Department of Cardiology and Pneumology, Universitätsmedizin Göttingen, Germany; DZHK (German Center for Cardiovascular Research), Partner Site Göttingen, Germany.Department of Cardiology and Pneumology, Universitätsmedizin Göttingen, Germany; Institute of Pharmacology and Toxicology, Technische Universität Dresden, Germany. Electronic address: kaomei.guan@tu-dresden.de.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

28941705

Citation

Streckfuss-Bömeke, Katrin, et al. "Severe DCM Phenotype of Patient Harboring RBM20 Mutation S635A Can Be Modeled By Patient-specific Induced Pluripotent Stem Cell-derived Cardiomyocytes." Journal of Molecular and Cellular Cardiology, vol. 113, 2017, pp. 9-21.
Streckfuss-Bömeke K, Tiburcy M, Fomin A, et al. Severe DCM phenotype of patient harboring RBM20 mutation S635A can be modeled by patient-specific induced pluripotent stem cell-derived cardiomyocytes. J Mol Cell Cardiol. 2017;113:9-21.
Streckfuss-Bömeke, K., Tiburcy, M., Fomin, A., Luo, X., Li, W., Fischer, C., Özcelik, C., Perrot, A., Sossalla, S., Haas, J., Vidal, R. O., Rebs, S., Khadjeh, S., Meder, B., Bonn, S., Linke, W. A., Zimmermann, W. H., Hasenfuss, G., & Guan, K. (2017). Severe DCM phenotype of patient harboring RBM20 mutation S635A can be modeled by patient-specific induced pluripotent stem cell-derived cardiomyocytes. Journal of Molecular and Cellular Cardiology, 113, 9-21. https://doi.org/10.1016/j.yjmcc.2017.09.008
Streckfuss-Bömeke K, et al. Severe DCM Phenotype of Patient Harboring RBM20 Mutation S635A Can Be Modeled By Patient-specific Induced Pluripotent Stem Cell-derived Cardiomyocytes. J Mol Cell Cardiol. 2017;113:9-21. PubMed PMID: 28941705.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Severe DCM phenotype of patient harboring RBM20 mutation S635A can be modeled by patient-specific induced pluripotent stem cell-derived cardiomyocytes. AU - Streckfuss-Bömeke,Katrin, AU - Tiburcy,Malte, AU - Fomin,Andrey, AU - Luo,Xiaojing, AU - Li,Wener, AU - Fischer,Claudia, AU - Özcelik,Cemil, AU - Perrot,Andreas, AU - Sossalla,Samuel, AU - Haas,Jan, AU - Vidal,Ramon Oliveira, AU - Rebs,Sabine, AU - Khadjeh,Sara, AU - Meder,Benjamin, AU - Bonn,Stefan, AU - Linke,Wolfgang A, AU - Zimmermann,Wolfram-Hubertus, AU - Hasenfuss,Gerd, AU - Guan,Kaomei, Y1 - 2017/09/21/ PY - 2017/04/09/received PY - 2017/09/01/revised PY - 2017/09/19/accepted PY - 2017/9/25/pubmed PY - 2018/6/2/medline PY - 2017/9/25/entrez KW - Alternative splicing KW - Cardiomyocytes KW - Dilated cardiomyopathy (DCM) KW - Induced pluripotent stem cells (iPSCs) KW - RNA-binding motif protein 20 (RBM20) KW - Titin (TTN) SP - 9 EP - 21 JF - Journal of molecular and cellular cardiology JO - J. Mol. Cell. Cardiol. VL - 113 N2 - The ability to generate patient-specific induced pluripotent stem cells (iPSCs) provides a unique opportunity for modeling heart disease in vitro. In this study, we generated iPSCs from a patient with dilated cardiomyopathy (DCM) caused by a missense mutation S635A in RNA-binding motif protein 20 (RBM20) and investigated the functionality and cell biology of cardiomyocytes (CMs) derived from patient-specific iPSCs (RBM20-iPSCs). The RBM20-iPSC-CMs showed abnormal distribution of sarcomeric α-actinin and defective calcium handling compared to control-iPSC-CMs, suggesting disorganized myofilament structure and altered calcium machinery in CMs of the RBM20 patient. Engineered heart muscles (EHMs) from RBM20-iPSC-CMs showed that not only active force generation was impaired in RBM20-EHMs but also passive stress of the tissue was decreased, suggesting a higher visco-elasticity of RBM20-EHMs. Furthermore, we observed a reduced titin (TTN) N2B-isoform expression in RBM20-iPSC-CMs by demonstrating a reduction of exon skipping in the PEVK region of TTN and an inhibition of TTN isoform switch. In contrast, in control-iPSC-CMs both TTN isoforms N2B and N2BA were expressed, indicating that the TTN isoform switch occurs already during early cardiogenesis. Using next generation RNA sequencing, we mapped transcriptome and splicing target profiles of RBM20-iPSC-CMs and identified different cardiac gene networks in response to the analyzed RBM20 mutation in cardiac-specific processes. These findings shed the first light on molecular mechanisms of RBM20-dependent pathological cardiac remodeling leading to DCM. Our data demonstrate that iPSC-CMs coupled with EHMs provide a powerful tool for evaluating disease-relevant functional defects and for a deeper mechanistic understanding of alternative splicing-related cardiac diseases. SN - 1095-8584 UR - https://www.unboundmedicine.com/medline/citation/28941705/Severe_DCM_phenotype_of_patient_harboring_RBM20_mutation_S635A_can_be_modeled_by_patient_specific_induced_pluripotent_stem_cell_derived_cardiomyocytes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2828(17)30310-3 DB - PRIME DP - Unbound Medicine ER -