An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque
Recombinant adeno-associated virus (rAAV)rh74.MCK.GALGT2 is a muscle-specific gene therapy that is being developed to treat forms of muscular dystrophy. Here we report on an isolated limb infusion technique in a non-human primate model, where hindlimb blood flow is transiently isolated using balloon...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2018-09-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050118300561 |
| _version_ | 1818151736936759296 |
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| author | Rui Xu Ying Jia Deborah A. Zygmunt Megan L. Cramer Kelly E. Crowe Guohong Shao Agatha E. Maki Haley N. Guggenheim Benjamin C. Hood Danielle A. Griffin Ellyn Peterson Brad Bolon John P. Cheatham Sharon L. Cheatham Kevin M. Flanigan Louise R. Rodino-Klapac Louis G. Chicoine Paul T. Martin |
| author_facet | Rui Xu Ying Jia Deborah A. Zygmunt Megan L. Cramer Kelly E. Crowe Guohong Shao Agatha E. Maki Haley N. Guggenheim Benjamin C. Hood Danielle A. Griffin Ellyn Peterson Brad Bolon John P. Cheatham Sharon L. Cheatham Kevin M. Flanigan Louise R. Rodino-Klapac Louis G. Chicoine Paul T. Martin |
| author_sort | Rui Xu |
| collection | DOAJ |
| description | Recombinant adeno-associated virus (rAAV)rh74.MCK.GALGT2 is a muscle-specific gene therapy that is being developed to treat forms of muscular dystrophy. Here we report on an isolated limb infusion technique in a non-human primate model, where hindlimb blood flow is transiently isolated using balloon catheters to concentrate vector in targeted leg muscles. A bilateral dose of 2.5 × 1013 vector genomes (vg)/kg/limb was sufficient to induce GALGT2-induced glycosylation in 10%–60% of skeletal myofibers in all leg muscles examined. There was a 19-fold ± 6-fold average limb-wide increase in vector genomes per microgram genomic DNA at a bilateral dose of 2.5 × 1013 vg/kg/limb compared with a bilateral dose of 6 × 1012 vg/kg/limb. A unilateral dose of 6 × 1013 vg/kg/limb showed a 12- ± 3-fold increase in treated limb muscles compared to contralateral untreated limb muscles, which received vector only after release into the systemic circulation from the treated limb. Variability in AAV biodistribution between different segments of the same muscle was 125% ± 18% for any given dose, while variability between the same muscle for any given treatment dose was 45% ± 7%. These experiments demonstrate that treatment of muscles throughout the leg with rAAVrh74.MCK.GALGT2 can be accomplished safely using an isolated limb infusion technique, where balloon catheters transiently isolate the limb vasculature, but that intra- and inter-muscle transduction variability is a significant issue. Keywords: limb perfusion, gene therapy, Galgt2, muscular dystrophy |
| first_indexed | 2024-12-11T13:43:34Z |
| format | Article |
| id | doaj.art-f9bc6cfb8158407cb2dff2b604340149 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-11T13:43:34Z |
| publishDate | 2018-09-01 |
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| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-f9bc6cfb8158407cb2dff2b6043401492022-12-22T01:04:39ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012018-09-011089104An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus MacaqueRui Xu0Ying Jia1Deborah A. Zygmunt2Megan L. Cramer3Kelly E. Crowe4Guohong Shao5Agatha E. Maki6Haley N. Guggenheim7Benjamin C. Hood8Danielle A. Griffin9Ellyn Peterson10Brad Bolon11John P. Cheatham12Sharon L. Cheatham13Kevin M. Flanigan14Louise R. Rodino-Klapac15Louis G. Chicoine16Paul T. Martin17Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA; Graduate Program in Molecular, Cellular and Developmental Biology, The Ohio State University, Columbus, OH 43210, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA; Graduate Program in Molecular, Cellular and Developmental Biology, The Ohio State University, Columbus, OH 43210, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USAGEMPath Inc., Longmont, CO 80504, USADepartment of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USADepartment of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA; Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA; Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA; Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USACenter for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA; Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USA; Corresponding author: Paul T. Martin, Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA.Recombinant adeno-associated virus (rAAV)rh74.MCK.GALGT2 is a muscle-specific gene therapy that is being developed to treat forms of muscular dystrophy. Here we report on an isolated limb infusion technique in a non-human primate model, where hindlimb blood flow is transiently isolated using balloon catheters to concentrate vector in targeted leg muscles. A bilateral dose of 2.5 × 1013 vector genomes (vg)/kg/limb was sufficient to induce GALGT2-induced glycosylation in 10%–60% of skeletal myofibers in all leg muscles examined. There was a 19-fold ± 6-fold average limb-wide increase in vector genomes per microgram genomic DNA at a bilateral dose of 2.5 × 1013 vg/kg/limb compared with a bilateral dose of 6 × 1012 vg/kg/limb. A unilateral dose of 6 × 1013 vg/kg/limb showed a 12- ± 3-fold increase in treated limb muscles compared to contralateral untreated limb muscles, which received vector only after release into the systemic circulation from the treated limb. Variability in AAV biodistribution between different segments of the same muscle was 125% ± 18% for any given dose, while variability between the same muscle for any given treatment dose was 45% ± 7%. These experiments demonstrate that treatment of muscles throughout the leg with rAAVrh74.MCK.GALGT2 can be accomplished safely using an isolated limb infusion technique, where balloon catheters transiently isolate the limb vasculature, but that intra- and inter-muscle transduction variability is a significant issue. Keywords: limb perfusion, gene therapy, Galgt2, muscular dystrophyhttp://www.sciencedirect.com/science/article/pii/S2329050118300561 |
| spellingShingle | Rui Xu Ying Jia Deborah A. Zygmunt Megan L. Cramer Kelly E. Crowe Guohong Shao Agatha E. Maki Haley N. Guggenheim Benjamin C. Hood Danielle A. Griffin Ellyn Peterson Brad Bolon John P. Cheatham Sharon L. Cheatham Kevin M. Flanigan Louise R. Rodino-Klapac Louis G. Chicoine Paul T. Martin An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque Molecular Therapy: Methods & Clinical Development |
| title | An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque |
| title_full | An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque |
| title_fullStr | An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque |
| title_full_unstemmed | An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque |
| title_short | An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque |
| title_sort | isolated limb infusion method allows for broad distribution of raavrh74 mck galgt2 to leg skeletal muscles in the rhesus macaque |
| url | http://www.sciencedirect.com/science/article/pii/S2329050118300561 |
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