Evaluation of vascular delivery methodologies to enhance rAAV6-mediated gene transfer to canine striated musculature.
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.
Gregorevic
Paul
P
Department of Neurology, University of Washington, Seattle, Washington 98195-7720, USA.
Schultz
Brian R
BR
Allen
James M
JM
Halldorson
Jeffrey B
JB
Blankinship
Michael J
MJ
Meznarich
Norman A
NA
Kuhr
Christian S
CS
Doremus
Caitlin
C
Finn
Eric
E
Liggitt
Denny
D
Chamberlain
Jeffrey S
JS
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
D058851:GPI-Linked Proteins; D007166:Immunosuppressive Agents; D007527:Isoenzymes; D016572:Cyclosporine; D000469:Alkaline Phosphatase; C019839:alkaline phosphatase, placental; D009173:Mycophenolic Acid