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 oligonucl eotide transfer. We demonstrated a pressure-dependent uptake of fluorescein -labeled (FITC) oligonucleotide (ODN) in rabbit carotid arteries with preex isting neointimal hyperplasia, using nondistending intravascular delivery p ressures ranging from 0 to 760 mmHg. At an infusion pressure of 50 mmHg, 10 .5 +/- 5% of neointimal cell nuclei were positive for nuclear uptake of FIT C-ODN 4 days after transfection, With an infusion pressure of 760 mmHg, the transfection efficiency increased to 84.2 +/- 5.3% of neointimal cells, an d to 64.5 +/- 11.6 and 92.4 +/- 5.5% of medial and adventitial cells, respe ctively. Similar patterns of FITC-ODN uptake were seen in atherosclerotic i njured arteries. We also investigated the pressure-mediated delivery of pla smid DNA. Transfection of a luciferase expression plasmid, using an infusio n pressure of 760 mmHg, 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 mmHg. Luciferase expression was significantily high er in pressure-transfected injured atherosclerotic arteries (5467.3 +/- 104 7.6 fg/mg protein at 760 mmHg). Transfection of beta-galactosidase indicate d that significant transgene expression occurred in the neointima and media . These data indicate that this pressure-mediated transfection method yield s efficient oligonucleotide delivery and enhances transduction with plasmid DNA in normal as well as injured nonatherosclerotic or atherosclerotic art eries.