Relative persistence of AAV serotype 1 vector genomes in dystrophic muscle
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BioMed Central
Open Access
Short paper
Relative persistence of AAV serotype 1 vector genomes in dystrophic muscle Christina A Pacak1,2, Thomas Conlon1,2, Cathryn S Mah*1,3 and Barry J Byrne*1,2,3 Address: 1Powell Gene Therapy Center, University of Florida, Gainesville, FL, USA, 2Department of Pediatrics, University of Florida, Gainesville, FL, USA and 3Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida, Gainesville, FL, USA Email: Christina A Pacak - [email protected]; Thomas Conlon - [email protected]; Cathryn S Mah* - [email protected]; Barry J Byrne* - [email protected] * Corresponding authors
Published: 15 October 2008 Genetic Vaccines and Therapy 2008, 6:14
doi:10.1186/1479-0556-6-14
Received: 10 June 2008 Accepted: 15 October 2008
This article is available from: http://www.gvt-journal.com/content/6/1/14 © 2008 Pacak et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract The purpose of this study was to assess the behavior of pseudotyped recombinant adenoassociated virus type 1 (rAAV2/1) vector genomes in dystrophic skeletal muscle. A comparison was made between a therapeutic vector and a reporter vector by injecting the hindlimb in a mouse model of Limb Girdle Muscular Dystrophy Type 2D (LGMD-2D) prior to disease onset. We hypothesized that the therapeutic vector would establish long-term persistence through prevention of myofiber turnover. In contrast, the reporter vector genome copy number would diminish over time due to disease-associated muscle degradation. One day old alpha sarcoglycan knockout mice (sgca-/-) were injected with 1 × 1011 vector genomes of rAAV2/1-tMCK-sgca in one hindlimb and the same dose of rAAV2/1-tMCK-LacZ in the contra lateral hindlimb. Newborn mice are tolerant of the foreign transgene allowing for long-term expression of both the marker and the therapeutic gene in the null background. At 2 time-points following vector administration, hindlimb muscles were harvested and analyzed for LacZ or sarcoglycan expression. Our data demonstrate prolonged vector genome persistence in skeletal muscle from the hindlimbs injected with the therapeutic transgene as compared to hindlimbs injected with the reporter gene. We observed loss of vector genomes in skeletal muscles that were there were not protected by the benefits of therapeutic gene transfer. In comparison, the therapeutic vector expressing sarcoglycan led to reduction or elimination of myofiber loss. Mitigating the membrane instability inherent in dystrophic muscle was able to prolong the life of individual myofibers.
Findings Limb Girdle Muscular Dystrophy Type 2D (LGMD-2D) is an autosomal recessive disorder caused by mutations in the alpha sarcoglycan gene (sgca) and is the most prevalent of the sarcoglycanopathies;
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