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Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes

Duchenne muscular dystrophy (DMD)–associated cardiac diseases are emerging as a major cause of morbidity and mortality in DMD patients, and many therapies for treatment of skeletal muscle failed to improve cardiac function. The reprogramming of patients’ somatic cells into pluripotent stem cells, co...

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Autores principales: Zatti, Susi, Martewicz, Sebastian, Serena, Elena, Uno, Narumi, Giobbe, Giovanni, Kazuki, Yasuhiro, Oshimura, Mitsuo, Elvassore, Nicola
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4365821/
https://www.ncbi.nlm.nih.gov/pubmed/26015941
http://dx.doi.org/10.1038/mtm.2013.1
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author Zatti, Susi
Martewicz, Sebastian
Serena, Elena
Uno, Narumi
Giobbe, Giovanni
Kazuki, Yasuhiro
Oshimura, Mitsuo
Elvassore, Nicola
author_facet Zatti, Susi
Martewicz, Sebastian
Serena, Elena
Uno, Narumi
Giobbe, Giovanni
Kazuki, Yasuhiro
Oshimura, Mitsuo
Elvassore, Nicola
author_sort Zatti, Susi
collection PubMed
description Duchenne muscular dystrophy (DMD)–associated cardiac diseases are emerging as a major cause of morbidity and mortality in DMD patients, and many therapies for treatment of skeletal muscle failed to improve cardiac function. The reprogramming of patients’ somatic cells into pluripotent stem cells, combined with technologies for correcting the genetic defect, possesses great potential for the development of new treatments for genetic diseases. In this study, we obtained human cardiomyocytes from DMD patient–derived, induced pluripotent stem cells genetically corrected with a human artificial chromosome carrying the whole dystrophin genomic sequence. Stimulation by cytokines was combined with cell culturing on hydrogel with physiological stiffness, allowing an adhesion-dependent maturation and a proper dystrophin expression. The obtained cardiomyocytes showed remarkable sarcomeric organization of cardiac troponin T and α-actinin, expressed cardiac-specific markers, and displayed electrically induced calcium transients lasting less than 1 second. We demonstrated that the human artificial chromosome carrying the whole dystrophin genomic sequence is stably maintained throughout the cardiac differentiation process and that multiple promoters of the dystrophin gene are properly activated, driving expression of different isoforms. These dystrophic cardiomyocytes can be a valuable source for in vitro modeling of DMD-associated cardiac disease. Furthermore, the derivation of genetically corrected, patient-specific cardiomyocytes represents a step toward the development of innovative cell and gene therapy approaches for DMD.
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spelling pubmed-43658212015-05-26 Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes Zatti, Susi Martewicz, Sebastian Serena, Elena Uno, Narumi Giobbe, Giovanni Kazuki, Yasuhiro Oshimura, Mitsuo Elvassore, Nicola Mol Ther Methods Clin Dev Article Duchenne muscular dystrophy (DMD)–associated cardiac diseases are emerging as a major cause of morbidity and mortality in DMD patients, and many therapies for treatment of skeletal muscle failed to improve cardiac function. The reprogramming of patients’ somatic cells into pluripotent stem cells, combined with technologies for correcting the genetic defect, possesses great potential for the development of new treatments for genetic diseases. In this study, we obtained human cardiomyocytes from DMD patient–derived, induced pluripotent stem cells genetically corrected with a human artificial chromosome carrying the whole dystrophin genomic sequence. Stimulation by cytokines was combined with cell culturing on hydrogel with physiological stiffness, allowing an adhesion-dependent maturation and a proper dystrophin expression. The obtained cardiomyocytes showed remarkable sarcomeric organization of cardiac troponin T and α-actinin, expressed cardiac-specific markers, and displayed electrically induced calcium transients lasting less than 1 second. We demonstrated that the human artificial chromosome carrying the whole dystrophin genomic sequence is stably maintained throughout the cardiac differentiation process and that multiple promoters of the dystrophin gene are properly activated, driving expression of different isoforms. These dystrophic cardiomyocytes can be a valuable source for in vitro modeling of DMD-associated cardiac disease. Furthermore, the derivation of genetically corrected, patient-specific cardiomyocytes represents a step toward the development of innovative cell and gene therapy approaches for DMD. Nature Publishing Group 2014-01-08 /pmc/articles/PMC4365821/ /pubmed/26015941 http://dx.doi.org/10.1038/mtm.2013.1 Text en Copyright © 2014 American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-nd/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivative Works 3.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Article
Zatti, Susi
Martewicz, Sebastian
Serena, Elena
Uno, Narumi
Giobbe, Giovanni
Kazuki, Yasuhiro
Oshimura, Mitsuo
Elvassore, Nicola
Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes
title Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes
title_full Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes
title_fullStr Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes
title_full_unstemmed Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes
title_short Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient–derived cardiomyocytes
title_sort complete restoration of multiple dystrophin isoforms in genetically corrected duchenne muscular dystrophy patient–derived cardiomyocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4365821/
https://www.ncbi.nlm.nih.gov/pubmed/26015941
http://dx.doi.org/10.1038/mtm.2013.1
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