Controlled release of small interfering RNA targeting midkine attenuates intimal hyperplasia in vein grafts
Controlled release of small interfering RNA targeting midkine attenuates intimal hyperplasia in vein grafts
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DOI：
10.1016/j.jvs.2006.04.044
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发表时间：
2006-09-01
影响因子：
4.3
通讯作者：
Kadomatsu, Kenji
中科院分区：
文献类型：
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作者：
Banno, Hiroshi;Takei, Yoshifumi;Kadomatsu, Kenji
目的：内膜增生是静脉移植后通畅的主要障碍。尽管有各种各样的试验来预防它，但尚未建立用于临床使用的令人满意的治疗策略。然而，充分抑制内膜增生的早期阶段可能会防止这种长期进行性疾病。中期因子（MK）是一种肝素结合生长因子，最初被发现是视黄酸反应基因的产物。我们以前证明，MK缺陷小鼠表现出显着减少新生内膜形成的再狭窄模型，这是逆转全身MK管理。在这项研究中，我们评估了一种策略，使用小干扰RNA（siRNA）靶向MK作为治疗静脉移植失败。在日本白色家兔中制作了颈静脉-颈动脉间置静脉移植物，用于低流量条件。将小干扰RNA与去端胶原混合，施加于移植静脉的外壁。Cy 3缀合的稳定siRNA用于确认其稳定性和成功转移到静脉移植物壁中。术后4周评价新生内膜增生。结果：MK表达在术后第7天达高峰，第10天达高峰，第10天达高峰，第10天达高峰。Cy 3标记的siRNA的荧光甚至可以在术后7天在移植物壁中检测到。逐渐增加的表达的敲低是通过使用去端肽胶原在血管周围应用siRNA来实现的。与对照组相比，移植后28天的内膜-中膜比率和内膜厚度均减少> 90%。这种现象之前的显着减少炎症细胞招聘到血管壁和随后的细胞增殖MK siRNA处理grafts.Conclusions：这些结果表明，中期因子是一个候选的分子靶点，用于预防静脉移植失败。此外，对于siRNA的临床应用，一个单一的术中端胶原为基础的非病毒递送方法可能是一个可靠的方法，以实现siRNA在体内的最大功能。这一策略可能是一个有用的和实用的形式的基因治疗对人类静脉移植失败。
Objective: Intimal hyperplasia is a major obstacle to patency after vein grafting. Despite of a diverse array of trials to prevent it, a satisfactory therapeutic strategy for clinical use has not been established. However, sufficient inhibition of early stages of intimal hyperplasia may prevent this long-term progressive disease. Midkine (MK) is a heparin-binding growth factor that was originally discovered as the product of a retinoic acid-responsive gene. We previously demonstrated that MK-deficient mice exhibit a striking reduction of neointima formation in a restenosis model, which is reversed on systemic MK administration. In this study, we evaluated a strategy of using small interfering RNA (siRNA) targeting MK as a therapy for vein graft failure.Methods: We first made a highly effective siRNA to rabbit MK. Jugular vein-to-carotid artery interposition vein grafts, which are applied to a low flow condition, were made in Japanese white rabbits. Small interfering RNA mixed with atelocollagen was administrated to the external wall of grafted veins. Cy3-conjugated stabilized siRNA was used to confirm its stability and successful transfer into the vein graft wall. Neointimal hyperplasia was evaluated 4 weeks after the operation. The proliferation index and leukocyte infiltration were determined.Results: MK expression was induced and reached the maximum level 7 days after operation. Fluorescence of Cy3-labeled siRNA could be detected in the graft wall even 7 days after operation. Knockdown of the gradually increasing expression was achieved by perivascular application of siRNA using atelocollagen. The intima-media ratio and the intima thickness at 28 days after grafting were both reduced > 90% by this treatment compared with controls. This phenomenon was preceded by significant reductions of inflammatory cell recruitment to the vessel walls and subsequent cell proliferation in MK siRNA-treated grafts.Conclusions: These results suggest that midkine is a candidate molecular target for preventing vein graft failure. Furthermore, for clinical applications of siRNA, a single intraoperative atelocollagen-based nonviral delivery method could be a reliable approach to achieve maximal function of siRNA in vivo. This strategy may be a useful and practical form of gene therapy against human vein graft failure.