Mazur W, Ali N M, Grinstead W C, Schulz D G, Raizner A E, French B A
Department of Medicine, Methodist Hospital, Houston, Texas.
Coron Artery Dis. 1994 Sep;5(9):779-86.
Restenosis after coronary angioplasty might be prevented by locally delivered gene therapy in conjunction with percutaneous transluminal coronary angioplasty (PTCA), since this approach should provide a sustained source of therapeutic protein within the dilated lesion. However, the potential application of gene therapy is limited by the technical barrier of efficiently transferring genes to vascular cells.
We used cultured coronary smooth muscle cells of human, porcine, and canine origin to evaluate three methods of gene transfer: recombinant adenovirus, liposomal complexes (Lipofectin), and Lipofectin supplemented with hemagglutinin. We then compared Lipofectin- and adenovirus-mediated direct gene transfer in canine and porcine coronary arteries.
The lipofection of cultured smooth muscle cells was enhanced by adding hemagglutinin, yielding luciferase levels that were 631-fold (human), ninefold (porcine), and sevenfold (canine) higher than with Lipofectin alone. However, the recombinant adenovirus directed even higher levels of gene expression, yielding luciferase levels that were 113,000-fold (human), 450-fold (porcine), and 230-fold (canine) higher than with Lipofectin alone. After percutaneous transluminal local delivery to intact canine coronary arteries, the adenovirus produced 55 times more luciferase than did Lipofectin. In living porcine coronary arteries, adenovirus produced 95 times more luciferase than did Lipofectin.
Recombinant adenovirus produces far more recombinant protein than does Lipofectin after percutaneous transluminal direct gene transfer to canine and porcine coronary arteries. Adenoviral vectors may therefore prove useful in evaluating the potential of gene therapy in large animal models of coronary restenosis.
冠状动脉血管成形术后的再狭窄或许可通过局部基因治疗联合经皮腔内冠状动脉血管成形术（PTCA）来预防，因为这种方法应能在扩张的病变部位提供持续的治疗性蛋白质来源。然而，基因治疗的潜在应用受到将基因有效转移至血管细胞这一技术障碍的限制。
我们使用人、猪和犬来源的培养冠状动脉平滑肌细胞来评估三种基因转移方法：重组腺病毒、脂质体复合物（Lipofectin）以及添加了血凝素的Lipofectin。然后我们比较了Lipofectin和腺病毒介导的犬和猪冠状动脉直接基因转移。
添加血凝素可增强培养平滑肌细胞的脂质体转染，产生的荧光素酶水平比单独使用Lipofectin时分别高出631倍（人）、9倍（猪）和7倍（犬）。然而，重组腺病毒介导的基因表达水平更高，产生的荧光素酶水平比单独使用Lipofectin时分别高出113000倍（人）、450倍（猪）和230倍（犬）。经皮腔内局部递送至完整的犬冠状动脉后，腺病毒产生的荧光素酶比Lipofectin多55倍。在活体猪冠状动脉中，腺病毒产生的荧光素酶比Lipofectin多95倍。
经皮腔内直接基因转移至犬和猪冠状动脉后，重组腺病毒产生的重组蛋白比Lipofectin多得多。因此，腺病毒载体可能在评估冠状动脉再狭窄大型动物模型中基因治疗的潜力方面有用。
