Percutaneous transluminal in vivo gene transfer by recombinant adenovirus in normal porcine coronary arteries, atherosclerotic arteries, and two models of coronary restenosis.
Percutaneous transluminal in vivo gene transfer by recombinant adenovirus in normal porcine coronary arteries, atherosclerotic arteries, and two models of coronary restenosis.
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DOI：
10.1161/01.cir.90.5.2402
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发表时间：
1994-11
期刊：
影响因子：
37.8
通讯作者：
B. A. French;Wojciech Mazur;N. Ali;R. Geske;J. Finnigan;G. Rodgers;R. Roberts;A. Raizner
中科院分区：
文献类型：
--
作者：
B. A. French;Wojciech Mazur;N. Ali;R. Geske;J. Finnigan;G. Rodgers;R. Roberts;A. Raizner
基因治疗已被提出作为冠状动脉血管成形术后再狭窄问题的可能解决方案。目前的研究是为了评估传统的基因转移方法，并开发经皮技术，将基因直接导入大型哺乳动物的冠状动脉。由于预期的靶向基因治疗再狭窄包括动脉粥样硬化和以前的仪器动脉，我们还评估了基因转移在动脉粥样硬化冠状动脉和两个猪模型的再狭窄：一个使用冠状动脉内支架和第二个使用球囊过度拉伸血管成形术。方法和结果使用穿孔球囊导管将Lipofectin-DNA复合物直接递送到完整动物的冠状动脉中的常规方法应用于18条猪冠状动脉，包括正常动脉、高胆固醇血症动脉和模拟再狭窄的动脉。本研究的结果与先前发表的结果一致，表明通过Lipofectin介导的基因转移只能获得低水平的荧光素酶基因表达。因此，我们进行了第二个平行的研究，以评估经皮腔内体内基因转移使用的复制缺陷型腺病毒载体。两项研究的比较显示，在接受腺病毒感染的队列中报告基因表达的平均水平比接受脂质转染的队列高100倍。正常动脉中荧光素酶活性随时间变化的分析显示，重组基因表达在1天后达到半最大值，在1周内达到峰值，在2周时仍为半最大值，并在4周时下降至低水平。用表达核定位β-半乳糖苷酶基因的第二种腺病毒处理的冠状动脉的组织化学分析表明，基因转移到中膜和外膜中有限数量的细胞中。使用针对CD44的单克隆抗体对Ad5输注的动脉进行免疫组织化学分析，鉴定出由白细胞组成的外膜周浸润。结论重组腺病毒载体在将外源基因直接导入哺乳动物冠状动脉方面比Lipofectin更有效。此外，高胆固醇血症和动脉损伤的影响似乎对使用任一方法获得的基因表达水平几乎没有影响。结果表明，低水平的重组基因表达，阻碍基因治疗预防再狭窄的主要障碍，可以通过使用腺病毒载体介导冠状动脉基因转移体内克服。这些载体提供的基因表达的持续时间和它们在动脉粥样硬化、球囊过度拉伸和支架冠状动脉中的有效部署表明，重组腺病毒可能具有在临床信息丰富的猪冠状动脉再狭窄模型中评估基因治疗的潜力。
BACKGROUND Gene therapy has been proposed as a possible solution to the problem of restenosis after coronary angioplasty. The current study was undertaken to assess conventional methods of gene transfer and to develop percutaneous techniques for introducing genes directly into the coronary arteries of large mammals. Since the anticipated targets of gene therapy against restenosis include atherosclerotic and previously instrumented arteries, we also evaluated gene transfer in atherosclerotic coronary arteries and in two porcine models of restenosis: one using intracoronary stents and a second using balloon overstretch angioplasty. METHODS AND RESULTS The conventional method of using perforated balloon catheters to deliver Lipofectin-DNA complexes directly into the coronary arteries of intact animals was applied to 18 porcine coronary arteries including normal arteries, hypercholesterolemic arteries, and those simulating restenosis. The results of this study were consistent with previously published results indicating that only low levels of luciferase gene expression could be obtained by Lipofectin-mediated gene transfer. We therefore undertook a second, parallel study to evaluate percutaneous transluminal in vivo gene transfer using a replication-deficient adenoviral vector. A comparison of the two studies revealed that the mean level of reporter gene expression in the cohort undergoing adenoviral infection was 100-fold higher than in the cohort undergoing Lipofection. Analysis of luciferase activity over time in normal arteries revealed that recombinant gene expression was half-maximal after 1 day, peaked within 1 week, was still half-maximal at 2 weeks, and declined to low levels by 4 weeks. Histochemical analysis of coronary arteries treated with a second adenovirus expressing a nuclear-localized beta-galactosidase gene demonstrated gene transfer to a limited number of cells in the media and adventitia. Immunohistochemical analysis of Ad5-infused arteries using a monoclonal antibody directed against CD44 identified a periadventitial infiltrate composed of leukocytes. CONCLUSIONS The recombinant adenoviral vectors proved to be far more effective than Lipofectin at delivering foreign genes directly into the coronary arteries of living mammals. Furthermore, the influences of hypercholesterolemia and arterial injury appeared to have little effect on the levels of gene expression obtained using either method. The results demonstrate that low-level recombinant gene expression, the major obstacle impeding gene therapy for the prevention of restenosis, can potentially be overcome by using adenoviral vectors to mediate coronary gene transfer in vivo. The duration of gene expression provided by these vectors and their effective deployment in atherosclerotic, balloon-overstretched, and stented coronary arteries suggest that recombinant adenovirus may have potential for evaluating gene therapy in the clinically informative porcine models of coronary restenosis.