A pressure-mediated nonviral method for efficient arterial gene and oligonucleotide transfer.
In this study, we report a method of controlled pressure-mediated delivery of "naked" DNA that achieves efficient and safe arterial gene and oligonucleotide transfer. We demonstrated a pressure-dependent uptake of fluorescein-labeled (FITC) oligonucleotide (ODN) in rabbit carotid arteries with preexisting neointimal hyperplasia, using nondistending intravascular delivery pressures ranging from 0 to 760 mm Hg. At an infusion pressure of 50 mm Hg, 10.5+/-5% of neointimal cell nuclei were positive for nuclear uptake of FITC-ODN 4 days after transfection. With an infusion pressure of 760 mm Hg, the transfection efficiency increased to 84.2+/-5.3% of neointimal cells, and to 64.5+/-11.6 and 92.4+/-5.5% of medial and adventitial cells, respectively. Similar patterns of FITC-ODN uptake were seen in atherosclerotic injured arteries. We also investigated the pressure-mediated delivery of plasmid DNA. Transfection of a luciferase expression plasmid, using an infusion pressure of 760 mm Hg, yielded luciferase expression of 816.6+/-108.6 fg/mg protein in normal rabbit carotid arteries, as compared with 38.9+/-23.7 fg/mg protein at 100 mm Hg. Luciferase expression was significantly higher in pressure-transfected injured atherosclerotic arteries (5467.3+/-1047.6 fg/mg protein at 760 mm Hg). Transfection of beta-galactosidase indicated that significant transgene expression occurred in the neointima and media. These data indicate that this pressure-mediated transfection method yields efficient oligonucleotide delivery and enhances transduction with plasmid DNA in normal as well as injured nonatherosclerotic or atherosclerotic arteries.
von der Leyen
H E
HE
Division of Cardiovascular Medicine, Falk Cardiovascular Research Center, Stanford University School of Medicine, CA 94305, USA.
Braun-Dullaeus
R
R
Mann
M J
MJ
Zhang
L
L
Niebauer
J
J
Dzau
V J
VJ
D000818:Animals; D001157:Arterial Occlusive Diseases / Q000473:pathology; D001161:Arteriosclerosis / Q000473:pathology; D002339:Carotid Arteries / Q000201:enzymology / Q000473:pathology; D002340:Carotid Artery Diseases / Q000473:pathology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D007150:Immunohistochemistry; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008365:Manometry; D008856:Microscopy, Fluorescence; D009841:Oligonucleotides; D010957:Plasmids; D011312:Pressure; D011817:Rabbits; D014162:Transfection / Q000379:methods; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D009841:Oligonucleotides; D008156:Luciferases; D001616:beta-Galactosidase