Gene delivery to pig coronary arteries from stents carrying antibody-tethered adenovirus
Gene delivery to pig coronary arteries from stents carrying antibody-tethered adenovirus
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DOI：
10.1089/10430340252792576
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发表时间：
2002-02-01
影响因子：
4.2
通讯作者：
Levy, RJ
中科院分区：
文献类型：
--
作者：
Klugherz, BD;Song, CX;Levy, RJ
冠状动脉支架的部署，以减轻动脉粥样硬化阻塞已受益于数以百万计的患者。然而，基因治疗，以防止支架内再狭窄，虽然在实验研究中有希望，仍然是一个挑战。用于病毒载体施用的常规策略利用递送病毒悬浮液输注的导管，这导致载体的次优定位和潜在危险的远端扩散。基于支架的基因递送可以避免这个问题。我们假设腺病毒基因载体从支架的位点特异性递送可以通过涉及抗病毒抗体栓系的机制来实现。用胶原蛋白涂层配制支架。抗腺病毒单克隆抗体共价结合到胶原蛋白表面。这些抗体能够通过高度特异性的抗原-抗体亲和力来束缚复制缺陷型腺病毒。我们首次报道了使用编码绿色荧光蛋白（GFP）的抗体栓系腺病毒成功进行基于支架的基因递送，证明了在细胞培养物和猪冠状动脉中向动脉平滑肌细胞的高效和高度定位的基因递送。猪的总动脉壁转导效率为总细胞的5.9 +/-1.1%。然而，新生内膜转导超过该区域总细胞的17%。重要的是，当使用特异性抗体拴系腺病毒时，在远端器官或支架动脉的下游段中，通过PCR检测不到载体的远端扩散。对照腺病毒支架，与非特异性抗体加腺病毒，表现出只有几个孤立的转导病灶，和穷人的位点特异性转导与载体的远端扩散。我们的结论是，血管支架是一个合适的平台，可定位的病毒载体输送系统，也防止系统传播的载体。使用载体的基于支架的抗病毒抗体栓系的基因递送应该适用于广泛的单一或多个治疗基因策略。
Deployment of coronary stents to relieve atherosclerotic obstruction has benefitted millions of patients. However, gene therapy to prevent in-stent restenosis, while promising in experimental studies, remains a challenge. Conventional strategies for viral vector administration utilize catheters that deliver infusions of viral suspensions, which result in suboptimal localization and potentially dangerous distal spread of vector. Stent-based gene delivery may circumvent this problem. We hypothesized that site-specific delivery of adenoviral gene vectors from a stent could be achieved through a mechanism involving anti-viral antibody tethering. Stents were formulated with a collagen coating. Anti-adenoviral monoclonal antibodies were covalently bound to the collagen surface. These antibodies enabled tethering of replication defective adenoviruses through highly specific antigen-antibody affinity. We report for the first time successful stent-based gene delivery using antibody-tethered adenovirus encoding green fluorescent protein (GFP), demonstrating efficient and highly localized gene delivery to arterial smooth muscle cells in both cell culture and pig coronary arteries. Overall arterial wall transduction efficiency in pigs was 5.9 +/- 1.1% of total cells. However, neointimal transduction was more than 17% of total cells in this region. Importantly, when specific antibody was used to tether adenovirus, no distal spread of vector was detectable by PCR, in either distal organs, or in the downstream segments of the stented arteries. Control adenovirus stents, with nonspecific antibody plus adenovirus, demonstrated only a few isolated foci of transduction, and poor site-specific transduction with distal spread of vector. We conclude that a vascular stent is a suitable platform for a localizable viral vector delivery system that also prevents systemic spread of vector. Gene delivery using stent-based anti-viral antibody tethering of vectors should be suitable for a wide array of single or multiple therapeutic gene strategies.