title: Complete restoration of multiple dystrophin isoforms in genetically corrected Duchenne muscular dystrophy patient-derived cardiomyocytes.
abstract: Duchenne muscular dystrophy (DMD)-associated cardiac diseases are emerging as a major cause of morbidity and mortality in DMD patients, and many therapies for treatment of skeletal muscle failed to improve cardiac function. The reprogramming of patients' somatic cells into pluripotent stem cells, combined with technologies for correcting the genetic defect, possesses great potential for the development of new treatments for genetic diseases. In this study, we obtained human cardiomyocytes from DMD patient-derived, induced pluripotent stem cells genetically corrected with a human artificial chromosome carrying the whole dystrophin genomic sequence. Stimulation by cytokines was combined with cell culturing on hydrogel with physiological stiffness, allowing an adhesion-dependent maturation and a proper dystrophin expression. The obtained cardiomyocytes showed remarkable sarcomeric organization of cardiac troponin T and α-actinin, expressed cardiac-specific markers, and displayed electrically induced calcium transients lasting less than 1 second. We demonstrated that the human artificial chromosome carrying the whole dystrophin genomic sequence is stably maintained throughout the cardiac differentiation process and that multiple promoters of the dystrophin gene are properly activated, driving expression of different isoforms. These dystrophic cardiomyocytes can be a valuable source for in vitro modeling of DMD-associated cardiac disease. Furthermore, the derivation of genetically corrected, patient-specific cardiomyocytes represents a step toward the development of innovative cell and gene therapy approaches for DMD.
authors.affiliation: Department of Industrial Engineering, University of Padova , Padova, Italy ; Venetian Institute of Molecular Medicine (VIMM) , Padova, Italy.
authors.forename: Susi
authors.initials: S
authors.lastname: Zatti
authors.affiliation: Department of Industrial Engineering, University of Padova , Padova, Italy ; Venetian Institute of Molecular Medicine (VIMM) , Padova, Italy.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Martewicz
authors.affiliation: Department of Industrial Engineering, University of Padova , Padova, Italy ; Venetian Institute of Molecular Medicine (VIMM) , Padova, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Serena
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University , Yonago, Japan.
authors.forename: Narumi
authors.initials: N
authors.lastname: Uno
authors.affiliation: Department of Industrial Engineering, University of Padova , Padova, Italy ; Venetian Institute of Molecular Medicine (VIMM) , Padova, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Giobbe
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University , Yonago, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kazuki
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University , Yonago, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Oshimura
authors.affiliation: Department of Industrial Engineering, University of Padova , Padova, Italy ; Venetian Institute of Molecular Medicine (VIMM) , Padova, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Elvassore