An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque

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

Molecular Therapy: Methods & Clinical Development (Sep 2018)

An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque

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

Affiliations

Rui Xu: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Ying Jia: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Deborah A. Zygmunt: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Megan L. Cramer: Center 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, USA
Kelly E. Crowe: Center 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, USA
Guohong Shao: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Agatha E. Maki: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Haley N. Guggenheim: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Benjamin C. Hood: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Danielle A. Griffin: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Ellyn Peterson: Center for Gene Therapy, The Research Institute at Nationwide Children’s Hospital, 700 Children’s Drive, Columbus, OH 43205, USA
Brad Bolon: GEMPath Inc., Longmont, CO 80504, USA
John P. Cheatham: Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USA
Sharon L. Cheatham: Department of Pediatrics, The Ohio State University College of Medicine, Columbus, OH 43210, USA
Kevin M. Flanigan: Center 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
Louise R. Rodino-Klapac: Center 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
Louis G. Chicoine: Center 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
Paul T. Martin: Center 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.

Journal volume & issue: Vol. 10
pp. 89 – 104

Abstract

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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

Published in Molecular Therapy: Methods & Clinical Development

ISSN: 2329-0501 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Biology (General): Genetics; Science: Biology (General): Cytology
Website: http://www.cell.com/molecular-therapy-family/methods/latest-content

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