• A Moretti, L Fonteyne, F Giesert, P Hoppmann, A B Meier, T Bozoglu, A Baehr, C M Schneider, D Sinnecker, K Klett, T Fröhlich, F Abdel Rahman, T Haufe, S Sun, V Jurisch, B Kessler, R Hinkel, R Dirschinger, E Martens, C Jilek, A Graf, S Krebs, G Santamaria, M Kurome, V Zakhartchenko, B Campbell, K Voelse, A Wolf, T Ziegler, S Reichert, S Lee, F Flenkenthaler, T Dorn, I Jeremias, H Blum, A Dendorfer, A Schnieke, S Krause, M C Walter, N Klymiuk, K L Laugwitz, E Wolf, W Wurst, and C Kupatt.
• Klinik und Poliklinik für Innere Medizin I, Klinikum rechts der Isar, Technical University Munich and German Center for Cardiovascular Research (DZHK), Munich Heart Alliance, Munich, Germany. amoretti@mytum.de.
• Nat. Med. 2020 Feb 1; 26 (2): 207-214.

AbstractFrameshift mutations in the DMD gene, encoding dystrophin, cause Duchenne muscular dystrophy (DMD), leading to terminal muscle and heart failure in patients. Somatic gene editing by sequence-specific nucleases offers new options for restoring the DMD reading frame, resulting in expression of a shortened but largely functional dystrophin protein. Here, we validated this approach in a pig model of DMD lacking exon 52 of DMD (DMDΔ52), as well as in a corresponding patient-derived induced pluripotent stem cell model. In DMDΔ52 pigs1, intramuscular injection of adeno-associated viral vectors of serotype 9 carrying an intein-split Cas9 (ref. 2) and a pair of guide RNAs targeting sequences flanking exon 51 (AAV9-Cas9-gE51) induced expression of a shortened dystrophin (DMDΔ51-52) and improved skeletal muscle function. Moreover, systemic application of AAV9-Cas9-gE51 led to widespread dystrophin expression in muscle, including diaphragm and heart, prolonging survival and reducing arrhythmogenic vulnerability. Similarly, in induced pluripotent stem cell-derived myoblasts and cardiomyocytes of a patient lacking DMDΔ52, AAV6-Cas9-g51-mediated excision of exon 51 restored dystrophin expression and amelioreate skeletal myotube formation as well as abnormal cardiomyocyte Ca2+ handling and arrhythmogenic susceptibility. The ability of Cas9-mediated exon excision to improve DMD pathology in these translational models paves the way for new treatment approaches in patients with this devastating disease.

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