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Absence of full-length dystrophin impairs normal maturation and contraction of cardiomyocytes derived from human-induced pluripotent stem cells.
Cardiovasc Res. 2020 02 01; 116(2):368-382.CR

Abstract

AIMS

Heart failure invariably affects patients with various forms of muscular dystrophy (MD), but the onset and molecular sequelae of altered structure and function resulting from full-length dystrophin (Dp427) deficiency in MD heart tissue are poorly understood. To better understand the role of dystrophin in cardiomyocyte development and the earliest phase of Duchenne muscular dystrophy (DMD) cardiomyopathy, we studied human cardiomyocytes differentiated from induced pluripotent stem cells (hiPSC-CMs) obtained from the urine of a DMD patient.

METHODS AND RESULTS

The contractile properties of patient-specific hiPSC-CMs, with no detectable dystrophin (DMD-CMs with a deletion of exon 50), were compared to CMs containing a CRISPR-Cas9 mediated deletion of a single G base at position 263 of the dystrophin gene (c.263delG-CMs) isogenic to the parental line of hiPSC-CMs from a healthy individual. We hypothesized that the absence of a dystrophin-actin linkage would adversely affect myofibril and cardiomyocyte structure and function. Cardiomyocyte maturation was driven by culturing long-term (80-100 days) on a nanopatterned surface, which resulted in hiPSC-CMs with adult-like dimensions and aligned myofibrils.

CONCLUSIONS

Our data demonstrate that lack of Dp427 results in reduced myofibril contractile tension, slower relaxation kinetics, and to Ca2+ handling abnormalities, similar to DMD cells, suggesting either retarded or altered maturation of cardiomyocyte structures associated with these functions. This study offers new insights into the functional consequences of Dp427 deficiency at an early stage of cardiomyocyte development in both patient-derived and CRISPR-generated models of dystrophin deficiency.

Authors+Show Affiliations

Experimental and Clinical Medicine, Div. of Physiology, University of Florence, Florence, Italy.Bioengineering, University of Washington, Seattle, WA, USA.Bioengineering, University of Washington, Seattle, WA, USA.School of Biosciences, University of Kent, Canterbury, UK.Pathology, University of Washington, Seattle, WA, USA. Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, USA.Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA.Bioengineering, University of Washington, Seattle, WA, USA.Experimental and Clinical Medicine, Div. of Physiology, University of Florence, Florence, Italy.Institute for Systems Biology, Seattle, Washington, USA.Institute for Systems Biology, Seattle, Washington, USA.Bioengineering, University of Washington, Seattle, WA, USA. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, USA.Experimental and Clinical Medicine, Div. of Physiology, University of Florence, Florence, Italy.Experimental and Clinical Medicine, Div. of Physiology, University of Florence, Florence, Italy.Bioengineering, University of Washington, Seattle, WA, USA. Pathology, University of Washington, Seattle, WA, USA. Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, USA.Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, USA. Rehabilitation Medicine, University of Washington, Seattle, WA, USA.Bioengineering, University of Washington, Seattle, WA, USA. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, USA. Rehabilitation Medicine, University of Washington, Seattle, WA, USA.Bioengineering, University of Washington, Seattle, WA, USA. Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

31049579

Citation

Pioner, J Manuel, et al. "Absence of Full-length Dystrophin Impairs Normal Maturation and Contraction of Cardiomyocytes Derived From Human-induced Pluripotent Stem Cells." Cardiovascular Research, vol. 116, no. 2, 2020, pp. 368-382.
Pioner JM, Guan X, Klaiman JM, et al. Absence of full-length dystrophin impairs normal maturation and contraction of cardiomyocytes derived from human-induced pluripotent stem cells. Cardiovasc Res. 2020;116(2):368-382.
Pioner, J. M., Guan, X., Klaiman, J. M., Racca, A. W., Pabon, L., Muskheli, V., Macadangdang, J., Ferrantini, C., Hoopmann, M. R., Moritz, R. L., Kim, D. H., Tesi, C., Poggesi, C., Murry, C. E., Childers, M. K., Mack, D. L., & Regnier, M. (2020). Absence of full-length dystrophin impairs normal maturation and contraction of cardiomyocytes derived from human-induced pluripotent stem cells. Cardiovascular Research, 116(2), 368-382. https://doi.org/10.1093/cvr/cvz109
Pioner JM, et al. Absence of Full-length Dystrophin Impairs Normal Maturation and Contraction of Cardiomyocytes Derived From Human-induced Pluripotent Stem Cells. Cardiovasc Res. 2020 02 1;116(2):368-382. PubMed PMID: 31049579.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Absence of full-length dystrophin impairs normal maturation and contraction of cardiomyocytes derived from human-induced pluripotent stem cells. AU - Pioner,J Manuel, AU - Guan,Xuan, AU - Klaiman,Jordan M, AU - Racca,Alice W, AU - Pabon,Lil, AU - Muskheli,Veronica, AU - Macadangdang,Jesse, AU - Ferrantini,Cecilia, AU - Hoopmann,Michael R, AU - Moritz,Robert L, AU - Kim,Deok-Ho, AU - Tesi,Chiara, AU - Poggesi,Corrado, AU - Murry,Charles E, AU - Childers,Martin K, AU - Mack,David L, AU - Regnier,Michael, PY - 2019/02/11/received PY - 2019/03/20/revised PY - 2019/04/17/accepted PY - 2019/5/3/pubmed PY - 2020/9/1/medline PY - 2019/5/4/entrez KW - CRISPR-Cas9 genome editing KW - Dystrophin KW - Human iPSC-cardiomyocytes KW - Muscular dystrophy KW - Myofibrils SP - 368 EP - 382 JF - Cardiovascular research JO - Cardiovasc. Res. VL - 116 IS - 2 N2 - AIMS: Heart failure invariably affects patients with various forms of muscular dystrophy (MD), but the onset and molecular sequelae of altered structure and function resulting from full-length dystrophin (Dp427) deficiency in MD heart tissue are poorly understood. To better understand the role of dystrophin in cardiomyocyte development and the earliest phase of Duchenne muscular dystrophy (DMD) cardiomyopathy, we studied human cardiomyocytes differentiated from induced pluripotent stem cells (hiPSC-CMs) obtained from the urine of a DMD patient. METHODS AND RESULTS: The contractile properties of patient-specific hiPSC-CMs, with no detectable dystrophin (DMD-CMs with a deletion of exon 50), were compared to CMs containing a CRISPR-Cas9 mediated deletion of a single G base at position 263 of the dystrophin gene (c.263delG-CMs) isogenic to the parental line of hiPSC-CMs from a healthy individual. We hypothesized that the absence of a dystrophin-actin linkage would adversely affect myofibril and cardiomyocyte structure and function. Cardiomyocyte maturation was driven by culturing long-term (80-100 days) on a nanopatterned surface, which resulted in hiPSC-CMs with adult-like dimensions and aligned myofibrils. CONCLUSIONS: Our data demonstrate that lack of Dp427 results in reduced myofibril contractile tension, slower relaxation kinetics, and to Ca2+ handling abnormalities, similar to DMD cells, suggesting either retarded or altered maturation of cardiomyocyte structures associated with these functions. This study offers new insights into the functional consequences of Dp427 deficiency at an early stage of cardiomyocyte development in both patient-derived and CRISPR-generated models of dystrophin deficiency. SN - 1755-3245 UR - https://www.unboundmedicine.com/medline/citation/31049579/Absence_of_full_length_dystrophin_impairs_normal_maturation_and_contraction_of_cardiomyocytes_derived_from_human_induced_pluripotent_stem_cells_ L2 - https://academic.oup.com/cardiovascres/article-lookup/doi/10.1093/cvr/cvz109 DB - PRIME DP - Unbound Medicine ER -