title: Cardiac Niche Influences the Direct Reprogramming of Canine Fibroblasts into Cardiomyocyte-Like Cells.
abstract: The Duchenne and Becker muscular dystrophies are caused by mutation of dystrophin gene and primarily affect skeletal and cardiac muscles. Cardiac involvement in dystrophic GRMD dogs has been demonstrated by electrocardiographic studies with the onset of a progressive cardiomyopathy similar to the cardiac disease in DMD patients. In this respect, GRMD is a useful model to explore cardiac and skeletal muscle pathogenesis and for developing new therapeutic protocols. Here we describe a protocol to convert GRMD canine fibroblasts isolated from heart and skin into induced cardiac-like myocytes (ciCLMs). We used a mix of transcription factors (GATA4, HAND2, TBX5, and MEF2C), known to be able to differentiate mouse and human somatic cells into ciCLMs. Exogenous gene expression was obtained using four lentiviral vectors carrying transcription factor genes and different resistance genes. Our data demonstrate a direct switch from fibroblast into ciCLMs with no activation of early cardiac genes. ciCLMs were unable to contract spontaneously, suggesting, differently from mouse and human cells, an incomplete differentiation process. However, when transplanted in neonatal hearts of SCID/Beige mice, ciCLMs participate in cardiac myogenesis.
authors.affiliation: Translational Cardiomyology Laboratory, Stem Cell Biology and Embryology Unit, Department of Development and Regeneration, KU Leuven, Herestraat 49 O&N4 Bus 814, 3000 Leuven, Belgium ; Laboratory of Stem Cells for Tissue Engineering, IRCCS, San Donato Hospital, San Donato, Milan, Italy.
authors.forename: Giacomo
authors.initials: G
authors.lastname: Palazzolo
authors.affiliation: Translational Cardiomyology Laboratory, Stem Cell Biology and Embryology Unit, Department of Development and Regeneration, KU Leuven, Herestraat 49 O&N4 Bus 814, 3000 Leuven, Belgium.
authors.forename: Mattia
authors.initials: M
authors.lastname: Quattrocelli
authors.affiliation: Organ Systems Development and Regeneration Unit, Department of Development and Regeneration, KU Leuven, Herestraat 49 O&N4 Bus 814, 3000 Leuven, Belgium.
authors.forename: Jaan
authors.initials: J
authors.lastname: Toelen
authors.affiliation: Laboratory of Clinical Chemistry, Public Health Corporation of Legnano, Magenta Hospital, 20013 Magenta, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Dominici
authors.affiliation: Laboratory of Stem Cells for Tissue Engineering, IRCCS, San Donato Hospital, San Donato, Milan, Italy ; Department of Biomedical Sciences for Health, University of Milan, Segrate, Milan, Italy.
authors.forename: Luigi
authors.identifier: 0000-0002-0712-2161
authors.initials: L
authors.lastname: Anastasia
authors.affiliation: Laboratory of Stem Cells for Tissue Engineering, IRCCS, San Donato Hospital, San Donato, Milan, Italy.
authors.forename: Guido
authors.initials: G
authors.lastname: Tettamenti
authors.affiliation: Laboratoire de Neurobiologie, Ecole Nationale Vétérinaire d'Alfort, 94704 Maisons-Alfort, France.
authors.forename: Inès
authors.initials: I
authors.lastname: Barthelemy
authors.affiliation: Laboratoire de Neurobiologie, Ecole Nationale Vétérinaire d'Alfort, 94704 Maisons-Alfort, France.
authors.forename: Stephane
authors.initials: S
authors.lastname: Blot
authors.affiliation: Laboratory of Molecular Virology and Gene Therapy, Department of Pharmaceutical and Pharmacological Sciences and Leuven Viral Vector Core, KU Leuven, Leuven, Belgium.
authors.forename: Rik
authors.initials: R
authors.lastname: Gijsbers
authors.affiliation: School of Life Sciences, EPFL, 1015 Lausanne, Switzerland.
authors.forename: Marco
authors.initials: M
authors.lastname: Cassano
authors.affiliation: Translational Cardiomyology Laboratory, Stem Cell Biology and Embryology Unit, Department of Development and Regeneration, KU Leuven, Herestraat 49 O&N4 Bus 814, 3000 Leuven, Belgium ; Human Anatomy Unit, Department of Public Health, Experimental and Forensic Medicine, University of Pavia, Via Forlanini 8, 27100 Pavia, Italy.
authors.forename: Maurilio
authors.identifier: 0000-0002-2422-3757
authors.initials: M
authors.lastname: Sampaolesi