Efficient Gene Transfer and Durable Transgene Expression in Grafted Rabbit Veins
Efficient Gene Transfer and Durable Transgene Expression in Grafted Rabbit Veins
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DOI：
10.1089/hum.2014.084
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发表时间：
2015-01-01
影响因子：
4.2
通讯作者：
Dichek, David A.
中科院分区：
文献类型：
--
作者：
Du, Liang;Zhang, Jingwan;Dichek, David A.
静脉搭桥术是治疗阻塞性冠状动脉疾病的有效方法。然而，它们的有效性受到动脉粥样硬化加速的限制。预防动脉粥样硬化的静脉搭桥基因工程可以改善移植物的长期通畅性和临床结果。我们使用兔颈静脉-颈动脉间置移植模型来开发移植静脉动脉粥样硬化的基因治疗。用第一代腺病毒载体可以很容易地原位转导兔静脉；然而，大多数转基因表达(接近80%)在动脉移植后3天内就消失了。这种转基因表达的快速丧失不能通过移植后的静脉转导或延长体外转导来阻止。然而，将静脉移植转导推迟28天(在静脉适应动脉循环之后)可以防止转基因表达的早期丧失。我们使用延迟转导方法来测试从辅助依赖腺病毒(HDAd)载体表达的治疗性转基因(载脂蛋白A-I)的持久性。HDAd DNA和载脂蛋白A-I mRNA在转导的静脉移植物中容易检测到。载体DNA和信使核糖核酸在4周后下降，并持续稳定至少6个月。移植后延迟28天的转导可以使移植静脉在基因转移前开始新生内膜生长和中膜增厚。然而，由于移植静脉逐渐向外重塑，移植静脉的管腔直径并未受到影响。我们的数据强调了HDAd介导的基因治疗在动脉化静脉移植物中用于预防静脉移植物动脉粥样硬化的前景。
Venous bypass grafts are useful treatments for obstructive coronary artery disease. However, their usefulness is limited by accelerated atherosclerosis. Genetic engineering of venous bypass grafts that prevented atherosclerosis could improve long-term graft patency and clinical outcomes. We used a rabbit model of jugular vein-to-carotid interposition grafting to develop gene therapy for vein-graft atherosclerosis. Rabbit veins were easily transduced in situ with a first-generation adenoviral vector; however, most transgene expression (similar to 80%) was lost within 3 days after arterial grafting. This rapid loss of transgene expression was not prevented by transducing veins after grafting or by prolonged ex vivo transduction. However, delaying vein-graft transduction for 28 days (after the vein had adapted to the arterial circulation) prevented this early loss of transgene expression. We used the delayed transduction approach to test the durability of expression of a therapeutic transgene (apolipoprotein A-I) expressed from a helper-dependent adenoviral (HDAd) vector. HDAd DNA and apolipoprotein A-I mRNA were easily detectable in transduced vein grafts. Vector DNA and mRNA declined by 4 weeks, and then persisted stably for at least 6 months. Delaying transduction for 28 days after grafting permitted initiation of vein-graft neointimal growth and medial thickening before gene transfer. However, vein-graft lumen diameter was not compromised, because of gradual outward remodeling of grafted veins. Our data highlight the promise of HDAd-mediated gene therapy, delivered to arterialized vein grafts, for preventing vein-graft atherosclerosis.