title: Duchenne Muscular Dystrophy Myogenic Cells from Urine-Derived Stem Cells Recapitulate the Dystrophin Genotype and Phenotype.
abstract: A ready source of autologous myogenic cells is of vital importance for drug screening and functional genetic studies in Duchenne muscular dystrophy (DMD), a rare disease caused by a variety of dystrophin gene mutations. As stem cells (SCs) can be easily and noninvasively obtained from urine specimens, we set out to determine whether they could be myogenically induced and useful in DMD research. To this end, we isolated stem cells from the urine of two healthy donors and from one patient with DMD, and performed surface marker characterization, myogenic differentiation (MyoD), and then transfection with antisense oligoribonucleotides to test for exon skipping and protein restoration. We demonstrated that native urine-derived stem cells express the full-length dystrophin transcript, and that the dystrophin mutation was retained in the cells of the patient with DMD, although the dystrophin protein was detected solely in control cells after myogenic transformation according to the phenotype. Notably, we also showed that treatment with antisense oligoribonucleotide against dystrophin exon 44 induced skipping in both native and MyoD-transformed urine-derived stem cells in DMD, with a therapeutic transcript-reframing effect, as well as visible protein restoration in the latter. Hence MyoD-transformed cells may be a good myogenic model for studying dystrophin gene expression, and native urine stem cells could be used to study the dystrophin transcript, and both diagnostic procedures and splicing modulation therapies in both patients and control subjects, without invasive and costly collection methods. New, bankable bioproducts from urine stem cells, useful for prescreening studies and therapeutic applications alike, are also foreseeable after further, more in-depth characterization.
authors.affiliation: 1 UOL (Unità Operativa Logistica) of Medical Genetics, University of Ferrara , Ferrara, Italy.
authors.forename: Maria Sofia
authors.initials: MS
authors.lastname: Falzarano
authors.affiliation: 2 Department of Cardiovascular Medicine, Catholic University of the Sacred Heart , Rome, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: D'Amario
authors.affiliation: 2 Department of Cardiovascular Medicine, Catholic University of the Sacred Heart , Rome, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Siracusano
authors.affiliation: 2 Department of Cardiovascular Medicine, Catholic University of the Sacred Heart , Rome, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Massetti
authors.affiliation: 3 Department of Cardiothoracic Surgery, Ospedale Bambino Gesù , Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Amodeo
authors.affiliation: 2 Department of Cardiovascular Medicine, Catholic University of the Sacred Heart , Rome, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: La Neve
authors.affiliation: 2 Department of Cardiovascular Medicine, Catholic University of the Sacred Heart , Rome, Italy.
authors.forename: Camilla Reina
authors.initials: CR
authors.lastname: Maroni
authors.affiliation: 4 Pediatric Neurology Unit, Catholic University, Rome , Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Mercuri
authors.affiliation: 1 UOL (Unità Operativa Logistica) of Medical Genetics, University of Ferrara , Ferrara, Italy.
authors.forename: Hana
authors.initials: H
authors.lastname: Osman
authors.affiliation: 1 UOL (Unità Operativa Logistica) of Medical Genetics, University of Ferrara , Ferrara, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Scotton
authors.affiliation: 1 UOL (Unità Operativa Logistica) of Medical Genetics, University of Ferrara , Ferrara, Italy.
authors.forename: Annarita
authors.initials: A
authors.lastname: Armaroli
authors.affiliation: 1 UOL (Unità Operativa Logistica) of Medical Genetics, University of Ferrara , Ferrara, Italy.
authors.forename: Rachele
authors.initials: R
authors.lastname: Rossi
authors.affiliation: 1 UOL (Unità Operativa Logistica) of Medical Genetics, University of Ferrara , Ferrara, Italy.
authors.forename: Rita
authors.initials: R
authors.lastname: Selvatici
authors.affiliation: 2 Department of Cardiovascular Medicine, Catholic University of the Sacred Heart , Rome, Italy.
authors.forename: Filippo
authors.initials: F
authors.lastname: Crea
authors.affiliation: 1 UOL (Unità Operativa Logistica) of Medical Genetics, University of Ferrara , Ferrara, Italy.|5 Neuromuscular Unit, Great Ormond Street Hospital, University College London , London, United Kingdom .
authors.forename: Alessandra
authors.initials: A
authors.lastname: Ferlini
mesh_terms: D016189:Dystrophin / Q000235:genetics*; D005091:Exons / Q000235:genetics; D015316:Genetic Therapy; D005838:Genotype; D006801:Humans; D024510:Muscle Development / Q000235:genetics; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000627:therapeutic use*; D010641:Phenotype; D013234:Stem Cells / Q000378:metabolism
chemical_list: D016189:Dystrophin; D016376:Oligonucleotides, Antisense