Evaluation of MYBPC3 trans-splicing and gene replacement as therapeutic options in human iPSC-Derived cardiomyocytes

• Verfasst von: Prondzynski, Maksymilian [VerfasserIn]
• Verfasst von: Müller, Oliver J. [VerfasserIn]
• Titel: Evaluation of MYBPC3 trans-splicing and gene replacement as therapeutic options in human iPSC-Derived cardiomyocytes
• Verf.angabe: Maksymilian Prondzynski, Elisabeth Krämer, Sandra D. Laufer, Aya Shibamiya, Ole Pless, Frederik Flenner, Oliver J. Müller, Julia Münch, Charles Redwood, Arne Hansen, Monica Patten, Thomas Eschenhagen, Giulia Mearini, Lucie Carrier
• E-Jahr: 2017
• Jahr: 16 June 2017
• Umfang: 12 S.
• Fussnoten: Gesehen am 09.04.2018
• Titel Quelle: Enthalten in: Molecular Therapy / Nucleic Acids
• Ort Quelle: New York, NY : Nature Publ. Group, 2012
• Jahr Quelle: 2017
• Band/Heft Quelle: 7(2017), Seite 475-486
• ISSN Quelle: 2162-2531
• DOI: doi:10.1016/j.omtn.2017.05.008
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: -splicing
• Sach-SW: gene replacement
• Sach-SW: human induced pluripotent stem cell-derived cardiomyocytes
• Sach-SW: hypertrophic cardiomyopathy
• K10plus-PPN: 1571801855

Abstract

Gene therapy is a promising option for severe forms of genetic diseases. We previously provided evidence for the feasibility of trans-splicing, exon skipping, and gene replacement in a mouse model of hypertrophic cardiomyopathy (HCM) carrying a mutation in MYBPC3, encoding cardiac myosin-binding protein C (cMyBP-C). Here we used human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) from an HCM patient carrying a heterozygous c.1358-1359insC MYBPC3 mutation and from a healthy donor. HCM hiPSC-CMs exhibited ∼50% lower MYBPC3 mRNA and cMyBP-C protein levels than control, no truncated cMyBP-C, larger cell size, and altered gene expression, thus reproducing human HCM features. We evaluated RNA trans-splicing and gene replacement after transducing hiPSC-CMs with adeno-associated virus. trans-splicing with 5′ or 3′ pre-trans-splicing molecules represented ∼1% of total MYBPC3 transcripts in healthy hiPSC-CMs. In contrast, gene replacement with the full-length MYBPC3 cDNA resulted in ∼2.5-fold higher MYBPC3 mRNA levels in HCM and control hiPSC-CMs. This restored the cMyBP-C level to 81% of the control level, suppressed hypertrophy, and partially restored gene expression to control level in HCM cells. This study provides evidence for (1) the feasibility of trans-splicing, although with low efficiency, and (2) efficient gene replacement in hiPSC-CMs with a MYBPC3 mutation.