title: Full-length dysferlin expression driven by engineered human dystrophic blood derived CD133+ stem cells.
abstract: The protein dysferlin is abundantly expressed in skeletal and cardiac muscles, where its main function is membrane repair. Mutations in the dysferlin gene are involved in two autosomal recessive muscular dystrophies: Miyoshi myopathy and limb-girdle muscular dystrophy type 2B. Development of effective therapies remains a great challenge. Strategies to repair the dysferlin gene by skipping mutated exons, using antisense oligonucleotides (AONs), may be suitable only for a subset of mutations, while cell and gene therapy can be extended to all mutations. AON-treated blood-derived CD133+ stem cells isolated from patients with Miyoshi myopathy led to partial dysferlin reconstitution in vitro but failed to express dysferlin after intramuscular transplantation into scid/blAJ dysferlin null mice. We thus extended these experiments producing the full-length dysferlin mediated by a lentiviral vector in blood-derived CD133+ stem cells isolated from the same patients. Transplantation of engineered blood-derived CD133+ stem cells into scid/blAJ mice resulted in sufficient dysferlin expression to correct functional deficits in skeletal muscle membrane repair. Our data suggest for the first time that lentivirus-mediated delivery of full-length dysferlin in stem cells isolated from Miyoshi myopathy patients could represent an alternative therapeutic approach for treatment of dysferlinopathies.
authors.affiliation: Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Centro Dino Ferrari, Milano, Italy.
authors.forename: Mirella
authors.initials: M
authors.lastname: Meregalli
authors.forename: Claire
authors.initials: C
authors.lastname: Navarro
authors.forename: Clementina
authors.initials: C
authors.lastname: Sitzia
authors.forename: Andrea
authors.initials: A
authors.lastname: Farini
authors.forename: Erica
authors.initials: E
authors.lastname: Montani
authors.forename: Nicolas
authors.initials: N
authors.lastname: Wein
authors.forename: Paola
authors.initials: P
authors.lastname: Razini
authors.forename: Cyriaque
authors.initials: C
authors.lastname: Beley
authors.forename: Letizia
authors.initials: L
authors.lastname: Cassinelli
authors.forename: Daniele
authors.initials: D
authors.lastname: Parolini
authors.forename: Marzia
authors.initials: M
authors.lastname: Belicchi
authors.forename: Dario
authors.initials: D
authors.lastname: Parazzoli
authors.forename: Luis
authors.initials: L
authors.lastname: Garcia
authors.forename: Yvan
authors.initials: Y
authors.lastname: Torrente
mesh_terms: D000071916:AC133 Antigen; D000328:Adult; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D049310:Distal Myopathies / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D000073939:Dysferlin; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D017404:In Situ Hybridization, Fluorescence; D007273:Injections, Intramuscular; D016086:Lentivirus / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008805:Mice, Inbred A; D016513:Mice, SCID; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000293:injuries / Q000378:metabolism / Q000473:pathology*; D009133:Muscular Atrophy / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D009154:Mutation / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010455:Peptides / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; C114839:DYSF protein, human; D000073939:Dysferlin; D006023:Glycoproteins; D008565:Membrane Proteins; D009124:Muscle Proteins; D016376:Oligonucleotides, Antisense; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D012333:RNA, Messenger
keywords: dysferlin; exon skipping; gene therapy; muscular dystrophy