title: Stem Cell-Mediated Exon Skipping of the Dystrophin Gene by the Bystander Effect.
abstract: Duchenne muscular dystrophy (DMD) is characterized by the loss of a functional dystrophin protein; the muscles of DMD patients progressively degenerate as a result of mechanical stress during contractions, and the condition eventually leads to premature death. By means antisense oligonucleotides (AONs), it is possible to modulate pre-mRNA splicing eliminating mutated exons and restoring dystrophin open reading frame. To overcome the hurdles in using AONs for therapeutic interventions, we exerted engineered human DMD stem cells with a lentivirus, which permanently and efficiently delivered the cloned AONs. Here we describe for the first time the exosome-mediated release of AONs from engineered human DMD CD133+ stem cells allowing the rescue of murine dystrophin expression. Finally, upon release, AONs could be internalized by host cells suggesting a potential role of exosomes acting as vesicular carriers for DMD gene therapy.
authors.forename: Mirella
authors.initials: M
authors.lastname: Meregalli
authors.forename: Andrea
authors.initials: A
authors.lastname: Farini
authors.forename: Clementina
authors.initials: C
authors.lastname: Sitzia
authors.forename: Cyriaque
authors.initials: C
authors.lastname: Beley
authors.forename: Paola
authors.initials: P
authors.lastname: Razini
authors.forename: Letizia
authors.initials: L
authors.lastname: Cassinelli
authors.forename: Federica
authors.initials: F
authors.lastname: Colleoni
authors.forename: Paola
authors.initials: P
authors.lastname: Frattini
authors.forename: Nadia
authors.initials: N
authors.lastname: Santo
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Galbiati
authors.forename: Davide
authors.initials: D
authors.lastname: Prosperi
authors.forename: Alessandro
authors.initials: A
authors.lastname: Tavelli
authors.forename: Marzia
authors.initials: M
authors.lastname: Belicchi
authors.forename: Luis
authors.initials: L
authors.lastname: Garcia
authors.affiliation: Laboratorio Cellule Staminali- Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti - Universita degli Studi di Milano Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Via F. Sforza 35, 20122 Milan, Italy. yvan.torrente@unimi.it.
authors.forename: Yvan
authors.initials: Y
authors.lastname: Torrente
mesh_terms: D000818:Animals; D024201:Bystander Effect / Q000502:physiology; D002478:Cells, Cultured; D016189:Dystrophin / Q000096:biosynthesis / Q000235:genetics*; D005091:Exons / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D018482:Muscle, Skeletal / Q000473:pathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics* / Q000628:therapy*; D016376:Oligonucleotides, Antisense / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics; D013234:Stem Cells / Q000166:cytology*
chemical_list: D016189:Dystrophin; D016376:Oligonucleotides, Antisense