Somatic gene editing ameliorates skeletal and cardiac muscle failure in pig and human models of Duchenne muscular dystrophy.

Moretti, A; Fonteyne, L; Giesert, F; Hoppmann, P; Meier, A B; Bozoglu, T; Baehr, A; Schneider, C M; Sinnecker, D; Klett, K; Fröhlich, T; Rahman, F Abdel; Haufe, T; Sun, S; Jurisch, V; Kessler, B; Hinkel, R; Dirschinger, R; Martens, E; Jilek, C; Graf, A; Krebs, S; Santamaria, G; Kurome, M; Zakhartchenko, V; Campbell, B; Voelse, K; Wolf, A; Ziegler, T; Reichert, S; Lee, S; Flenkenthaler, F; Dorn, T; Jeremias, I; Blum, H; Dendorfer, A; Schnieke, A; Krause, S; Walter, M C; Klymiuk, N; Laugwitz, K L; Wolf, E; Wurst, W; Kupatt, C     «

Moretti, A; Fonteyne, L; Giesert, F; Hoppmann, P; Meier, A B; Bozoglu, T; Baehr, A; Schneider, C M; Sinnecker, D; Klett, K; Fröhlich, T; Rahman, F Abdel; Haufe, T; Sun, S; Jurisch, V; Kessler, B; Hinkel, R; Dirschinger, R; Martens, E; Jilek, C; Graf, A; Krebs, S; Santamaria, G; Kurome, M; Zakhartchenko, V; Campbell, B; Voelse, K; Wolf, A; Ziegler, T; Reichert, S; Lee, S; Flenkenthaler, F; Dorn, T; Jeremias, I; Blum, H; Dendorfer, A; Schnieke, A; Krause, S; Walter, M C; Klymiuk, N; Laugwitz, K L;...     »

Frameshift 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.      «

Frameshift 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 pluripoten...     »