Optimization of regional intraarterial naked DNA-mediated transgene delivery to skeletal muscles in a large animal model.
Effective gene therapy for muscular dystrophy will likely require intravascular administration. Although plasmid DNA (pDNA) contained within a large volume and rapidly infused into a major artery can achieve gene transfer within downstream muscles, this is associated with substantial muscle edema. Here we hypothesized that excessive edema-related increases in intramuscular pressure (IM pressure) developed during intraarterial pDNA injections could hinder successful gene delivery. Accordingly, we monitored IM pressure during injection of pDNA carrying a LacZ transgene into the femoral artery of rats and pigs. Large variations in IM pressure were found between different muscles. There was a significant inverse relationship between IM pressure and the subsequent level of gene transfer to muscle. Modification of the injection protocol to reduce IM pressure led to greatly increased pDNA-mediated gene expression and reduced muscle damage in pigs. Under the most optimized conditions, average transfection within eight different muscles of the pig hind limb amounted to 22% of all fibers, attaining a maximum of 60% in the gastrocnemius muscle. We conclude that IM pressure monitoring is a simple and useful procedure, which can be applied in both small and large animals to help optimize pDNA-mediated gene transfer to skeletal muscles by the intraarterial route.
Danialou
Gawiyou
G
Meakins-Christie Laboratories and Respiratory Division, McGill University, Montreal, QC, Canada H3A 1A1.
Comtois
Alain S
AS
Matecki
Stefan
S
Nalbantoglu
Josephine
J
Karpati
George
G
Gilbert
Renald
R
Geoffroy
Pascale
P
Gilligan
Sandra
S
Tanguay
Jean-Francois
JF
Petrof
Basil J
BJ
D000818:Animals; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics* / Q000494:pharmacology; D007275:Injections, Intravenous; D008297:Male; D023421:Models, Animal; D018482:Muscle, Skeletal / Q000378:metabolism*; D011312:Pressure; D051381:Rats; D013552:Swine / Q000235:genetics; D019076:Transgenes / Q000235:genetics*
D004247:DNA