MRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy.

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Citation: Gene Therapy. 20(3):274-82, 2013 Mar.PMID: 22551778Institution: MedStar Heart & Vascular InstituteForm of publication: Journal ArticleMedline article type(s): Journal Article | Research Support, N.I.H., Intramural | Research Support, Non-U.S. Gov'tSubject headings: *Dependovirus/ge [Genetics] | *Dystrophin/ge [Genetics] | *Magnetic Resonance Imaging/mt [Methods] | *Muscular Dystrophy, Duchenne/th [Therapy] | *RNA, Small Nuclear/ge [Genetics] | *Transduction, Genetic/mt [Methods] | Animals | Base Sequence | Blotting, Western | Disease Models, Animal | Dogs | Dystrophin/me [Metabolism] | Exons/ge [Genetics] | Female | Gene Expression | Genetic Therapy/mt [Methods] | Genetic Vectors/ge [Genetics] | Humans | Immunohistochemistry | Male | Molecular Sequence Data | Muscular Dystrophy, Duchenne/ge [Genetics] | Muscular Dystrophy, Duchenne/me [Metabolism] | Myocardium/me [Metabolism] | Reverse Transcriptase Polymerase Chain Reaction | RNA, Small Nuclear/me [Metabolism] | Sf9 CellsYear: 2013ISSN:
  • 0969-7128
Name of journal: Gene therapyAbstract: Duchenne muscular dystrophy (DMD) cardiomyopathy patients currently have no therapeutic options. We evaluated catheter-based transendocardial delivery of a recombinant adeno-associated virus (rAAV) expressing a small nuclear U7 RNA (U7smOPT) complementary to specific cis-acting splicing signals. Eliminating specific exons restores the open reading frame resulting in translation of truncated dystrophin protein. To test this approach in a clinically relevant DMD model, golden retriever muscular dystrophy (GRMD) dogs received serotype 6 rAAV-U7smOPT via the intracoronary or transendocardial route. Transendocardial injections were administered with an injection-tipped catheter and fluoroscopic guidance using X-ray fused with magnetic resonance imaging (XFM) roadmaps. Three months after treatment, tissues were analyzed for DNA, RNA, dystrophin protein, and histology. Whereas intracoronary delivery did not result in effective transduction, transendocardial injections, XFM guidance, enabled 30+10 non-overlapping injections per animal. Vector DNA was detectable in all samples tested and ranged from <1 to >3000 vector genome copies per cell. RNA analysis, western blot analysis, and immunohistology demonstrated extensive expression of skipped RNA and dystrophin protein in the treated myocardium. Left ventricular function remained unchanged over a 3-month follow-up. These results demonstrated that effective transendocardial delivery of rAAV-U7smOPT was achieved using XFM. This approach restores an open reading frame for dystrophin in affected dogs and has potential clinical utility.All authors: Barbash IM, Bogan JR, Cecchini S, Faranesh AZ, Garcia L, Hoyt RF, Kornegay JN, Kotin RM, Lederman RJ, Li L, Virag T, Yang YFiscal year: FY2013Digital Object Identifier: Date added to catalog: 2014-04-22
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Journal Article MedStar Authors Catalog Article 22551778 Available 22551778

Duchenne muscular dystrophy (DMD) cardiomyopathy patients currently have no therapeutic options. We evaluated catheter-based transendocardial delivery of a recombinant adeno-associated virus (rAAV) expressing a small nuclear U7 RNA (U7smOPT) complementary to specific cis-acting splicing signals. Eliminating specific exons restores the open reading frame resulting in translation of truncated dystrophin protein. To test this approach in a clinically relevant DMD model, golden retriever muscular dystrophy (GRMD) dogs received serotype 6 rAAV-U7smOPT via the intracoronary or transendocardial route. Transendocardial injections were administered with an injection-tipped catheter and fluoroscopic guidance using X-ray fused with magnetic resonance imaging (XFM) roadmaps. Three months after treatment, tissues were analyzed for DNA, RNA, dystrophin protein, and histology. Whereas intracoronary delivery did not result in effective transduction, transendocardial injections, XFM guidance, enabled 30+10 non-overlapping injections per animal. Vector DNA was detectable in all samples tested and ranged from <1 to >3000 vector genome copies per cell. RNA analysis, western blot analysis, and immunohistology demonstrated extensive expression of skipped RNA and dystrophin protein in the treated myocardium. Left ventricular function remained unchanged over a 3-month follow-up. These results demonstrated that effective transendocardial delivery of rAAV-U7smOPT was achieved using XFM. This approach restores an open reading frame for dystrophin in affected dogs and has potential clinical utility.

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