Cationic polymer and lipids enhance adenovirus-mediated gene transfer to rabbit carotid artery.
OBJECTIVE
Improvement of efficiency of gene transfer to endothelium could be useful for several applications. We tested the hypothesis that cationic nonviral molecules augment adenovirus-mediated gene transfer to blood vessels, perhaps by alteration of the surface charge of adenovirus and facilitation of binding to endothelium.


METHODS
Carotid arteries from rabbits were incubated in vitro for 0.5 to 2 hours with an adenoviral vector alone or noncovalent complexes of adenovirus with poly-L-lysine (a cationic polymer) or lipofectin (a cationic lipid). Binding of adenovirus to the vessels was evaluated immediately after incubation with virus, and assay of transgene (ss-galactosidase) activity and histochemistry were performed 24 hours after gene transfer. To determine whether cationic molecules can be used to augment alteration of vascular function by adenovirus-mediated gene transfer, we also examined effects on gene transfer of endothelial nitric oxide synthase.


RESULTS
Assay of ss-galactosidase activity indicated that both cationic molecules increased transgene expression in vessels by approximately 5- to 6-fold. In contrast, when endothelium was removed from the vessels after gene transfer, poly-L-lysine and lipofectin did not significantly increase transgene activity. Histochemistry for ss-galactosidase also suggested that the adenovirus-cationic molecule complexes augmented transgene expression mainly in the endothelium. In addition, we found that complexing adenovirus with cationic molecules increased binding of adenovirus to the vessels. After gene transfer with recombinant adenovirus containing endothelial nitric oxide synthase, calcium ionophore (A23187) produced greater relaxation of vessels treated with adenovirus complexed with poly-L-lysine or lipofectin than those treated with adenovirus alone.


CONCLUSIONS
Cationic molecules improve the efficiency of adenovirus-mediated gene transfer to blood vessels.
Departments of Internal Medicine, Physiology and Biophysics, and Pharmacology, Cardiovascular Center and Center on Aging, University of Iowa College of Medicine, Iowa City, Iowa.
K
K
Toyoda
H
H
Ooboshi
Y
Y
Chu
A
A
Fasbender
B L
BL
Davidson
M J
MJ
Welsh
D D
DD
Heistad
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D002339:Carotid Arteries / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D004279:DNA, Viral / Q000378:metabolism; D018014:Gene Transfer Techniques; D006651:Histocytochemistry; D008163:Luminescent Measurements; D008297:Male; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D010714:Phosphatidylethanolamines / Q000494:pharmacology*; D011107:Polylysine / Q000494:pharmacology*; D011817:Rabbits; D014666:Vasomotor System / Q000187:drug effects / Q000502:physiology; D001616:beta-Galactosidase / Q000378:metabolism
D004279:DNA, Viral; D010714:Phosphatidylethanolamines; D011107:Polylysine; C020888:1,2-dielaidoylphosphatidylethanolamine; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D001616:beta-Galactosidase