Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy
RNA trans-splicing has been explored as a therapeutic option for a variety of genetic diseases, but not for cardiac genetic disease. Hypertrophic cardiomyopathy (HCM) is an autosomal-dominant disease, characterized by left ventricular hypertrophy (LVH) and diastolic dysfunction. MYBPC3, encoding car...
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doaj-71f14212fc7c4a9d880522573ae986ba2020-11-24T22:38:34ZengElsevierMolecular Therapy: Nucleic Acids2162-25312013-01-012C10.1038/mtna.2013.31Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic CardiomyopathyGiulia Mearini0Doreen Stimpel1Elisabeth Krämer2Birgit Geertz3Ingke Braren4Christina Gedicke-Hornung5Guillaume Précigout6Oliver J Müller7Hugo A Katus8Thomas Eschenhagen9Thomas Voit10Luis Garcia11Stéphanie Lorain12Lucie Carrier13Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyDepartment of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyDepartment of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyDepartment of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyDepartment of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyDepartment of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyUniversité Pierre et Marie Curie UM 76, Inserm U974, CNRS UMR7215, Institut de Myologie, Paris, FranceDepartment of Cardiology, Internal Medicine III, University Hospital Heidelberg, Heidelberg, GermanyDepartment of Cardiology, Internal Medicine III, University Hospital Heidelberg, Heidelberg, GermanyDepartment of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyUniversité Pierre et Marie Curie UM 76, Inserm U974, CNRS UMR7215, Institut de Myologie, Paris, FranceUniversité Pierre et Marie Curie UM 76, Inserm U974, CNRS UMR7215, Institut de Myologie, Paris, FranceUniversité Pierre et Marie Curie UM 76, Inserm U974, CNRS UMR7215, Institut de Myologie, Paris, FranceDepartment of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyRNA trans-splicing has been explored as a therapeutic option for a variety of genetic diseases, but not for cardiac genetic disease. Hypertrophic cardiomyopathy (HCM) is an autosomal-dominant disease, characterized by left ventricular hypertrophy (LVH) and diastolic dysfunction. MYBPC3, encoding cardiac myosin-binding protein C (cMyBP-C) is frequently mutated. We evaluated the 5′-trans-splicing strategy in a mouse model of HCM carrying a Mybpc3 mutation. 5′-trans-splicing was induced between two independently transcribed molecules, the mutant endogenous Mypbc3 pre-mRNA and an engineered pre-trans-splicing molecule (PTM) carrying a FLAG-tagged wild-type (WT) Mybpc3 cDNA sequence. PTMs were packaged into adeno-associated virus (AAV) for transduction of cultured cardiac myocytes and the heart in vivo. Full-length repaired Mybpc3 mRNA represented up to 66% of total Mybpc3 transcripts in cardiac myocytes and 0.14% in the heart. Repaired cMyBP-C protein was detected by immunoprecipitation in cells and in vivo and exhibited correct incorporation into the sarcomere in cardiac myocytes. This study provides (i) the first evidence of successful 5′-trans-splicing in vivo and (ii) proof-of-concept of mRNA repair in the most prevalent cardiac genetic disease. Since current therapeutic options for HCM only alleviate symptoms, these findings open new horizons for causal therapy of the severe forms of the disease.http://www.sciencedirect.com/science/article/pii/S2162253116301639hypertrophic cardiomyopathyMybpc3RNA-based therapytrans-splicing |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Giulia Mearini Doreen Stimpel Elisabeth Krämer Birgit Geertz Ingke Braren Christina Gedicke-Hornung Guillaume Précigout Oliver J Müller Hugo A Katus Thomas Eschenhagen Thomas Voit Luis Garcia Stéphanie Lorain Lucie Carrier |
spellingShingle |
Giulia Mearini Doreen Stimpel Elisabeth Krämer Birgit Geertz Ingke Braren Christina Gedicke-Hornung Guillaume Précigout Oliver J Müller Hugo A Katus Thomas Eschenhagen Thomas Voit Luis Garcia Stéphanie Lorain Lucie Carrier Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy Molecular Therapy: Nucleic Acids hypertrophic cardiomyopathy Mybpc3 RNA-based therapy trans-splicing |
author_facet |
Giulia Mearini Doreen Stimpel Elisabeth Krämer Birgit Geertz Ingke Braren Christina Gedicke-Hornung Guillaume Précigout Oliver J Müller Hugo A Katus Thomas Eschenhagen Thomas Voit Luis Garcia Stéphanie Lorain Lucie Carrier |
author_sort |
Giulia Mearini |
title |
Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy |
title_short |
Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy |
title_full |
Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy |
title_fullStr |
Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy |
title_full_unstemmed |
Repair of Mybpc3 mRNA by 5′-trans-splicing in a Mouse Model of Hypertrophic Cardiomyopathy |
title_sort |
repair of mybpc3 mrna by 5′-trans-splicing in a mouse model of hypertrophic cardiomyopathy |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2013-01-01 |
description |
RNA trans-splicing has been explored as a therapeutic option for a variety of genetic diseases, but not for cardiac genetic disease. Hypertrophic cardiomyopathy (HCM) is an autosomal-dominant disease, characterized by left ventricular hypertrophy (LVH) and diastolic dysfunction. MYBPC3, encoding cardiac myosin-binding protein C (cMyBP-C) is frequently mutated. We evaluated the 5′-trans-splicing strategy in a mouse model of HCM carrying a Mybpc3 mutation. 5′-trans-splicing was induced between two independently transcribed molecules, the mutant endogenous Mypbc3 pre-mRNA and an engineered pre-trans-splicing molecule (PTM) carrying a FLAG-tagged wild-type (WT) Mybpc3 cDNA sequence. PTMs were packaged into adeno-associated virus (AAV) for transduction of cultured cardiac myocytes and the heart in vivo. Full-length repaired Mybpc3 mRNA represented up to 66% of total Mybpc3 transcripts in cardiac myocytes and 0.14% in the heart. Repaired cMyBP-C protein was detected by immunoprecipitation in cells and in vivo and exhibited correct incorporation into the sarcomere in cardiac myocytes. This study provides (i) the first evidence of successful 5′-trans-splicing in vivo and (ii) proof-of-concept of mRNA repair in the most prevalent cardiac genetic disease. Since current therapeutic options for HCM only alleviate symptoms, these findings open new horizons for causal therapy of the severe forms of the disease. |
topic |
hypertrophic cardiomyopathy Mybpc3 RNA-based therapy trans-splicing |
url |
http://www.sciencedirect.com/science/article/pii/S2162253116301639 |
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