title: Evaluation of vascular delivery methodologies to enhance rAAV6-mediated gene transfer to canine striated musculature.
abstract: A growing body of research supports the development of recombinant adeno-associated viral (rAAV) vectors for delivery of gene expression cassettes to striated musculature as a method of treating severe neuromuscular conditions. However, it is unclear whether delivery protocols that achieve extensive gene transfer in mice can be adapted to produce similarly extensive gene transfer in larger mammals and ultimately patients. Consequently, we sought to investigate methodological modifications that would facilitate rAAV-mediated gene transfer to the striated musculature of canines. A simple procedure incorporating acute (i) occlusion of limb blood flow, (ii) exsanguination via compression bandage, and (iii) vector "dwell" time of <20 minutes, markedly enhanced the transduction of limb muscles, compared with a simple bolus limb infusion of vector. A complementary method whereby vector was infused into the jugular vein led to efficient transduction of cardiomyocytes and to a lesser degree the diaphragm. Together these methods can be used to achieve transgene expression in heart, diaphragm, and limb muscles of juvenile dogs using rAAV6 vectors. These results establish that rAAV-mediated gene delivery is a viable approach to achieving systemic transduction of striated musculature in mammals approaching the dimensions of newborn humans.
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195-7720, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Gregorevic
authors.forename: Brian R
authors.initials: BR
authors.lastname: Schultz
authors.forename: James M
authors.initials: JM
authors.lastname: Allen
authors.forename: Jeffrey B
authors.initials: JB
authors.lastname: Halldorson
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Blankinship
authors.forename: Norman A
authors.initials: NA
authors.lastname: Meznarich
authors.forename: Christian S
authors.initials: CS
authors.lastname: Kuhr
authors.forename: Caitlin
authors.initials: C
authors.lastname: Doremus
authors.forename: Eric
authors.initials: E
authors.lastname: Finn
authors.forename: Denny
authors.initials: D
authors.lastname: Liggitt
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Chamberlain
mesh_terms: D000469:Alkaline Phosphatase; D000818:Animals; D001808:Blood Vessels / Q000187:drug effects / Q000378:metabolism*; D016572:Cyclosporine / Q000494:pharmacology; D000229:Dependovirus / Q000235:genetics*; D003964:Diaphragm / Q000378:metabolism; D004285:Dogs; D058851:GPI-Linked Proteins; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D006614:Hindlimb / Q000378:metabolism; D006801:Humans; D007165:Immunosuppression Therapy; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics / Q000276:immunology; D007601:Jugular Veins / Q000378:metabolism; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D009173:Mycophenolic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D009206:Myocardium / Q000378:metabolism
chemical_list: D058851:GPI-Linked Proteins; D007166:Immunosuppressive Agents; D007527:Isoenzymes; D016572:Cyclosporine; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental; D009173:Mycophenolic Acid