title: Cationic PMMA nanoparticles bind and deliver antisense oligoribonucleotides allowing restoration of dystrophin expression in the mdx mouse.
abstract: For subsets of Duchenne muscular dystrophy (DMD) mutations, antisense oligoribonucleotide (AON)-mediated exon skipping has proven to be efficacious in restoring the expression of dystrophin protein. In the mdx murine model systemic delivery of AON, recognizing the splice donor of dystrophin exon 23, has shown proof of concept. Here, we show that using cationic polymethylmethacrylate (PMMA) (marked as T1) nanoparticles loaded with a low dose of 2'-O-methyl-phosphorothioate (2'OMePS) AON delivered by weekly intraperitoneal (IP) injection (0.9 mg/kg/week), could restore dystrophin expression in body-wide striated muscles. Delivery of an identical dose of naked AON did not result in detectable dystrophin expression. Transcription, western, and immunohistochemical analysis showed increased levels of dystrophin transcript and protein, and correct localization at the sarcolemma. This study shows that T1 nanoparticles have the capacity to bind and convoy AONs in body-wide muscle tissues and to reduce the dose required for dystrophin rescue. By immunofluorescence and electron microscopy studies, we highlighted the diffusion pathways of this compound. This nonviral approach may valuably improve the therapeutic usage of AONs in DMD as well as the delivery of RNA molecules with many implications in both basic research and medicine.
authors.affiliation: Department of Experimental and Diagnostic Medicine, Section of Medical Genetics, University of Ferrara, Ferrara, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Rimessi
authors.forename: Patrizia
authors.initials: P
authors.lastname: Sabatelli
authors.forename: Marina
authors.initials: M
authors.lastname: Fabris
authors.forename: Paola
authors.initials: P
authors.lastname: Braghetta
authors.forename: Elena
authors.initials: E
authors.lastname: Bassi
authors.forename: Pietro
authors.initials: P
authors.lastname: Spitali
authors.forename: Gaetano
authors.initials: G
authors.lastname: Vattemi
authors.forename: Giuliano
authors.initials: G
authors.lastname: Tomelleri
authors.forename: Lara
authors.initials: L
authors.lastname: Mari
authors.forename: Daniela
authors.initials: D
authors.lastname: Perrone
authors.forename: Alessandro
authors.initials: A
authors.lastname: Medici
authors.forename: Marcella
authors.initials: M
authors.lastname: Neri
authors.forename: Matteo
authors.initials: M
authors.lastname: Bovolenta
authors.forename: Elena
authors.initials: E
authors.lastname: Martoni
authors.forename: Nadir M
authors.initials: NM
authors.lastname: Maraldi
authors.forename: Francesca
authors.initials: F
authors.lastname: Gualandi
authors.forename: Luciano
authors.initials: L
authors.lastname: Merlini
authors.forename: Marco
authors.initials: M
authors.lastname: Ballestri
authors.forename: Luisa
authors.initials: L
authors.lastname: Tondelli
authors.forename: Katia
authors.initials: K
authors.lastname: Sparnacci
authors.forename: Paolo
authors.initials: P
authors.lastname: Bonaldo
authors.forename: Antonella
authors.initials: A
authors.lastname: Caputo
authors.forename: Michele
authors.initials: M
authors.lastname: Laus
authors.forename: Alessandra
authors.initials: A
authors.lastname: Ferlini
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016189:Dystrophin / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D018101:Mice, Inbred mdx; D008817:Mice, Mutant Strains; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D009137:Muscular Dystrophy, Animal / Q000235:genetics / Q000628:therapy; D053758:Nanoparticles / Q000737:chemistry*; D020320:Oligoribonucleotides, Antisense / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D019904:Polymethyl Methacrylate / Q000138:chemical synthesis / Q000737:chemistry*
chemical_list: D016189:Dystrophin; D020320:Oligoribonucleotides, Antisense; D019904:Polymethyl Methacrylate