Riessen R, Rahimizadeh H, Blessing E, Takeshita S, Barry J J, Isner J M
Department of Medicine (Cardiology), St. Elizabeth's Hospital, Tufts University School of Medicine, Boston, MA 02135.
Hum Gene Ther. 1993 Dec;4(6):749-58. doi: 10.1089/hum.1993.4.6-749.
Direct arterial gene transfer has been previously achieved using double-balloon catheters and perforated balloons, in most cases facilitated by the use of cationic liposomes or viral vectors. These gene delivery systems, however, have been compromised by issues relating to efficacy and/or safety, and furthermore require that angioplasty be performed independent of gene transfer. We investigated the possibility that arterial gene transfer might be performed during balloon angioplasty by delivery of naked genetic material from a thin coat of hydrogel polymer applied to a standard angioplasty balloon. Transfections with luciferase DNA applied to a hydrogel balloon were performed in rabbit arteries. Luciferase expression 3 days after transfection was tested in three different models: (i) an organ culture model (n = 10); (ii) surgically exposed carotid arteries (n = 14); and (iii) external iliac arteries using a percutaneous approach (n = 13). Supplementary transfections (n = 3), intended to identify the site of arterial transfection, were performed using the gene encoding for nuclear-specific beta-galactosidase (beta-gal). All rabbit arteries transfected with the luciferase gene (37/37; 100%) expressed luciferase activity. Gene expression achieved in vivo, either in the surgically exposed carotid arteries or in the external iliac arteries transfected percutaneously, was quantitatively similar to that achieved in the organ culture model. Reduction in the duration of inflation from 30 min to 1 min had no statistically significant impact on transfection efficiency. Gene expression was documented to persist up to 14 days post percutaneous transfection. Analysis of arteries transfected with nuclear-specific beta-gal showed the presence of the transgene in intimal and subintimal sites. These results demonstrate that vascular gene transfer can be performed successfully without liposomes or viral vectors using DNA applied to a standard angioplasty catheter balloon coated with hydrogel. Percutaneous transfection with a hydrogel-coated balloon permits gene transfer coincident with the angioplasty procedure itself, even with inflations as short as 1 min.
直接动脉基因转移此前已通过双球囊导管和多孔球囊实现，在大多数情况下借助阳离子脂质体或病毒载体来促进。然而，这些基因递送系统存在与疗效和/或安全性相关的问题，此外还要求血管成形术独立于基因转移进行。我们研究了在球囊血管成形术期间通过将裸露的遗传物质从应用于标准血管成形术球囊的水凝胶聚合物薄涂层中递送进行动脉基因转移的可能性。将荧光素酶DNA应用于水凝胶球囊，在兔动脉中进行转染。在三种不同模型中测试了转染后3天的荧光素酶表达：(i) 器官培养模型（n = 10）；(ii) 手术暴露的颈动脉（n = 14）；以及 (iii) 使用经皮方法的髂外动脉（n = 13）。使用编码核特异性β-半乳糖苷酶（β-gal）的基因进行补充转染（n = 3），以确定动脉转染的部位。所有用荧光素酶基因转染的兔动脉（37/37；100%）均表达荧光素酶活性。在体内，无论是在手术暴露的颈动脉还是经皮转染的髂外动脉中实现的基因表达，在定量上与在器官培养模型中实现的相似。将充气持续时间从30分钟减少到1分钟对转染效率没有统计学上的显著影响。经皮转染后基因表达被记录持续长达14天。对用核特异性β-gal转染的动脉进行分析显示，在内膜和内膜下部位存在转基因。这些结果表明，使用应用于涂有水凝胶的标准血管成形术导管球囊的DNA，无需脂质体或病毒载体即可成功进行血管基因转移。用水凝胶涂层球囊进行经皮转染允许基因转移与血管成形术本身同时进行，即使充气时间短至1分钟。
