The New Strategy of Gene Therapy for the Patients with Pheripheral Arterial Disease

血管成形术后的再狭窄和静脉旁路手术后内膜增生导致的移植物晚期失效是血管成形术的主要临床问题。我们以前已经表明，新生内膜形成显着抑制中期因子缺陷小鼠。如果给予缺陷小鼠中期因子蛋白，则新生内膜形成恢复。中期因子（MK）是一种肝素结合生长因子，参与炎症细胞和血管平滑肌细胞的迁移。在这里，我们评估MK反义寡脱氧核苷酸（ODN）预防再狭窄的潜力。此外，我们还对靶向MK的siRNA策略作为静脉移植失败的治疗进行了评估。1.生长因子MK反义寡核苷酸抑制球囊损伤诱导的新生内膜形成：我们克隆了兔MK的cDNA。球囊损伤诱导兔颈动脉MKe表达，最高水平发生在血管成形术后7-14天。反义ODN抑制 关于我们 2. MKe的siRNA控制释放对移植静脉内膜增生的抑制作用：术后7天，MK的表达达到最高水平。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了90%以上。结论：MK是治疗血管再狭窄的候选分子靶点。此外，MKe是预防静脉移植失败的候选分子靶点。此外，对于siRNA的临床应用，基于去端胶原的单一术中非病毒递送方法可能是实现siRNA在体内最大功能的可靠方法。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。少

Restenosis after angioplasty and late graft failure due to intimal hyperplasia after vein bypass surgery are the major clinical problem of angioplasty. We have previously shown that neointima formation is strikingly suppressed in midkine-deficient mice. Neointima formation is restored if midkine protein is administrated to the deficient mice. Midkine(MK) is a heparin-binding growth factor, and implicated in the migration of inflammatory cells and vascular smooth muscle cells. Here, we evaluated the potential of MK antisense oligodeoxyribonucleotide (ODN) for the prevention of restenosis. In addition, we evaluated the siRNA targeting MK strategy as a therapy for vein graft failure.1.Antisense Oligodeoxyribonucleotide as to the Growth Factor MK Suppresses Neointima Formation Induced by Balloon Injury : We cloned the cDNA of rabbit MKe. The balloon injury induced MKe expression, the maximum level occurring 7-14 days after angioplasty, in the rabbit carotid artery. The antisense ODN suppre … More ssed MK induction in vivo, and consequently suppressed neointima formation to 60% of the control level.2.Controlled Release of siRNA as to MKe Attenuates Intimal Hyperplasia in Vein Grafts : MK expression was induced and reached the maximum level 7 days afteroperation. Knockdown of the gradually increasing expression was achieved by perivascular application of siRNA using atelocollagen. Both the intima/media ratio and the intima thickness at 28 days after grafting were reduced by more than 90% by this treatment as compared with in controls. Conclusions : These results suggest that MK is a candidate molecular target for the therapy for vascular restenosis. In addition, MKe is a candidate molecular target for preventing vein graft failure.Furthermore, for clinical applications of siRNA, a single intraoperative atelocollagen-based non viral 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. Less