000			03785nam a22006017a 4500
008			140422s20132013 xxu||||| |||| 00| 0 eng d
022			_a0969-7128
040			_aOvid MEDLINE(R)
099			_a22551778
245			_aMRI roadmap-guided transendocardial delivery of exon-skipping recombinant adeno-associated virus restores dystrophin expression in a canine model of Duchenne muscular dystrophy.
251			_aGene Therapy. 20(3):274-82, 2013 Mar.
252			_aGene Ther. 20(3):274-82, 2013 Mar.
253			_aGene therapy
260			_c2013
260			_fFY2013
266			_d2014-04-22
520			_aDuchenne 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.
650			_a*Dependovirus/ge [Genetics]
650			_a*Dystrophin/ge [Genetics]
650			_a*Magnetic Resonance Imaging/mt [Methods]
650			_a*Muscular Dystrophy, Duchenne/th [Therapy]
650			_a*RNA, Small Nuclear/ge [Genetics]
650			_a*Transduction, Genetic/mt [Methods]
650			_aAnimals
650			_aBase Sequence
650			_aBlotting, Western
650			_aDisease Models, Animal
650			_aDogs
650			_aDystrophin/me [Metabolism]
650			_aExons/ge [Genetics]
650			_aFemale
650			_aGene Expression
650			_aGenetic Therapy/mt [Methods]
650			_aGenetic Vectors/ge [Genetics]
650			_aHumans
650			_aImmunohistochemistry
650			_aMale
650			_aMolecular Sequence Data
650			_aMuscular Dystrophy, Duchenne/ge [Genetics]
650			_aMuscular Dystrophy, Duchenne/me [Metabolism]
650			_aMyocardium/me [Metabolism]
650			_aReverse Transcriptase Polymerase Chain Reaction
650			_aRNA, Small Nuclear/me [Metabolism]
650			_aSf9 Cells
651			_aMedStar Heart & Vascular Institute
657			_aJournal Article
657			_aResearch Support, N.I.H., Intramural
657			_aResearch Support, Non-U.S. Gov't
700			_aBarbash, Israel M
790			_aBarbash IM, Bogan JR, Cecchini S, Faranesh AZ, Garcia L, Hoyt RF, Kornegay JN, Kotin RM, Lederman RJ, Li L, Virag T, Yang Y
856			_uhttp://dx.doi.org/10.1038/gt.2012.38 _zhttp://dx.doi.org/10.1038/gt.2012.38
942			_cART _dArticle
999			_c1071 _d1071