Percutaneous arterial gene transfer in a rabbit model. Efficiency in normal and balloon-dilated atherosclerotic arteries.

Abstract

The possibility of using an exclusively percutaneous strategy to deliver foreign DNA to normal and balloon-dilated atherosclerotic arteries was studied by analysis of transfection efficiency in a rabbit model. A total of 22 external iliac arteries from 22 rabbits (10 normal and 12 atherosclerotic) were transfected with a solution of luciferase expression vector plasmid and liposome, using a dual balloon-catheter system. Analysis of the transfected segments revealed luciferase activity in 10 of the 22 arteries (4/10 normal vs 6/12 balloon-injured atherosclerotic, P = NS); no activity could be detected in the contralateral limb arterial segments used as controls. Luciferase activity levels in successfully transfected segments measured 4.10±1.19 (m±SEM) Turner light units (TLU), with 3.03±1.16 TLU found in normals vs 4.81±1.87 TLU in balloon-injured atherosclerotic arteries (P = NS). In situ hybridization of successfully transfected atherosclerotic sections showed expression of the luciferase gene mRNA from rare cells (< 1 / 1,000) limited to the neointimal lesion. Thus, expression of new genetic material may be achieved in both normal and balloon-dilated atherosclerotic arteries following an exclusively percutaneous approach. The low efficiency ofthe current delivery strategy, however, represents a potential limitation that must be improved if this strategy is to be applied as a therapeutic approach to human vascular disease. (J. Clin. Invest. 1992.90:936-944.)

Low-efficiency of percutaneous adenovirus-mediated arterial gene transfer in the atherosclerotic rabbit.

“…In a few studies, the transgene, inserted into the adenoviral genome, has been successfully delivered to the arterial wall using a percutaneous approach similar to the procedure used to perform coronary angioplasty (4,5,7,9,10). Despite the variation in catheter designs used, the resulting transfection efficiencies have been typically higher than those achieved with retroviral vectors (11,12) plasmid DNA/liposome complexes (11,13,14), or plasmid DNA alone (15,16). It is unclear, however, whether these results, obtained in normal or endothelium-denuded arteries, can be extended to more extensively diseased arteries (17).…”

Imaging of light emission from the expression of luciferases in living cells and organisms: a review

“…Following liposome-mediated transformation of luciferse expression vectors, luciferase expression has been monitored in normal and atherosclerotic external iliac rabbit arteries (149). Replication-deficient adenovirusbearing firefly luciferase and b-galactosidase genes were successfully used to test the comparative efficiencies of transgenic gene delivery to cultured Sprague-Dawley rat thoracic artery and aortic artery smooth muscle tissue cultures (150).…”

Pre-treatment with elastase improves the efficiency of percutaneous adenovirus-mediated gene transfer to the arterial media

“…37,38 Briefly, 3 days following transfection the animals were killed by pentobarbital overdose. The iliac arteries were removed, washed in PBS at 4°C, then homogenized in 1 ml cell lysis reagent (Promega) using a Tekmar tissuemizer for 1 min.…”