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Evaluation of MYBPC3 trans-splicing and gene replacement as therapeutic options in human iPSC-Derived cardiomyocytes
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| Veröffentlicht in: | Molecular Therapy / Nucleic Acids 7(2017), Seite 475-486 |
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| Personen und Körperschaften: | , |
| Titel: | Evaluation of MYBPC3 trans-splicing and gene replacement as therapeutic options in human iPSC-Derived cardiomyocytes/ 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 |
| Format: | E-Book-Kapitel |
| Sprache: | Englisch |
| veröffentlicht: |
16 June 2017
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| Gesamtaufnahme: |
: Molecular Therapy / Nucleic Acids, 7(2017), Seite 475-486
, volume:7 |
| Schlagwörter: | |
| Quelle: | Verbunddaten SWB Lizenzfreie Online-Ressourcen |
| Zusammenfassung: | 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. |
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| Beschreibung: | Gesehen am 09.04.2018 |
| Umfang: | 12 |
| ISSN: |
2162-2531
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| DOI: | 10.1016/j.omtn.2017.05.008 |