title: Antisense-mediated exon skipping: a therapeutic strategy for titin-based dilated cardiomyopathy.
abstract: Frameshift mutations in the TTN gene encoding titin are a major cause for inherited forms of dilated cardiomyopathy (DCM), a heart disease characterized by ventricular dilatation, systolic dysfunction, and progressive heart failure. To date, there are no specific treatment options for DCM patients but heart transplantation. Here, we show the beneficial potential of reframing titin transcripts by antisense oligonucleotide (AON)-mediated exon skipping in human and murine models of DCM carrying a previously identified autosomal-dominant frameshift mutation in titin exon 326. Correction of TTN reading frame in patient-specific cardiomyocytes derived from induced pluripotent stem cells rescued defective myofibril assembly and stability and normalized the sarcomeric protein expression. AON treatment in Ttn knock-in mice improved sarcomere formation and contractile performance in homozygous embryos and prevented the development of the DCM phenotype in heterozygous animals. These results demonstrate that disruption of the titin reading frame due to a truncating DCM mutation can be restored by exon skipping in both patient cardiomyocytes in vitro and mouse heart in vivo, indicating RNA-based strategies as a potential treatment option for DCM.
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University, Tübingen, Germany Victor Chang Cardiac Research Institute, Darlinghurst, NSW, Australia michael.gramlich@med.uni-tuebingen.de amoretti@med1.med.tum.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Gramlich
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany.
authors.forename: Luna Simona
authors.initials: LS
authors.lastname: Pane
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University, Tübingen, Germany.
authors.forename: Qifeng
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany.
authors.forename: Zhifen
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany Department of Biomedical Sciences, University of Padova, Padua, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Murgia
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University, Tübingen, Germany.
authors.forename: Sonja
authors.initials: S
authors.lastname: Schötterl
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Goedel
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University, Tübingen, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Metzger
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Brade
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany Department of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, Catanzaro, Italy.
authors.forename: Elvira
authors.initials: E
authors.lastname: Parrotta
authors.affiliation: Department of Dermatology, Eberhard Karls University, Tübingen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Schaller
authors.affiliation: Libin Cardiovascular Institute of Alberta and University of Calgary, Calgary, AB, Canada.
authors.forename: Brenda
authors.initials: B
authors.lastname: Gerull
authors.affiliation: Max Delbrueck Center for Molecular Medicine, Berlin, Germany.
authors.forename: Ludwig
authors.initials: L
authors.lastname: Thierfelder
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Annemieke
authors.initials: A
authors.lastname: Aartsma-Rus
authors.affiliation: Institute for Integrative Pathophysiology, Universitätsmedizin Mannheim, Mannheim, Germany.
authors.forename: Siegfried
authors.initials: S
authors.lastname: Labeit
authors.affiliation: Department of Cardiology, Royal Brisbane and Women's Hospital and University of Queensland School of Medicine, Brisbane, Australia.
authors.forename: John J
authors.initials: JJ
authors.lastname: Atherton
authors.affiliation: Genetic Health Queensland, Royal Brisbane and Women's Hospital, Brisbane, Qld, Australia.
authors.forename: Julie
authors.initials: J
authors.lastname: McGaughran
authors.affiliation: Victor Chang Cardiac Research Institute, Darlinghurst, NSW, Australia St Vincent's Clinical School, University of New South Wales, Kensington, NSW, Australia.
authors.forename: Richard P
authors.initials: RP
authors.lastname: Harvey
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Sinnecker
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany DZHK (German Centre for Cardiovascular Research) - partner site Munich Heart Alliance, Munich, Germany.
authors.forename: Karl-Ludwig
authors.initials: KL
authors.lastname: Laugwitz
authors.affiliation: Department of Cardiology and Cardiovascular Diseases, Eberhard Karls University, Tübingen, Germany.
authors.forename: Meinrad Paul
authors.initials: MP
authors.lastname: Gawaz
authors.affiliation: I. Medical Department - Cardiology, Klinikum rechts der Isar - Technische Universität München, Munich, Germany DZHK (German Centre for Cardiovascular Research) - partner site Munich Heart Alliance, Munich, Germany michael.gramlich@med.uni-tuebingen.de amoretti@med1.med.tum.de.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Moretti
mesh_terms: D000818:Animals; D002311:Cardiomyopathy, Dilated / Q000503:physiopathology* / Q000628:therapy; D002478:Cells, Cultured; D064211:Connectin / Q000235:genetics / Q000378:metabolism*; D003584:Cytological Techniques; D004195:Disease Models, Animal; D005091:Exons; D016368:Frameshift Mutation; D005786:Gene Expression Regulation / Q000187:drug effects*; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000502:physiology; D009210:Myofibrils / Q000378:metabolism / Q000502:physiology; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use
chemical_list: D064211:Connectin; D016376:Oligonucleotides, Antisense
keywords: dilated cardiomyopathy; exon skipping; induced pluripotent stem cells; titin