Ultrasound-mediated delivery of TIMP-3 plasmid DNA into saphenous vein leads to increased lumen size in a porcine interposition graft model
Ultrasound-mediated delivery of TIMP-3 plasmid DNA into saphenous vein leads to increased lumen size in a porcine interposition graft model
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DOI：
10.1038/sj.gt.3302498
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发表时间：
2005-07-01
期刊：
影响因子：
5.1
通讯作者：
Newman, CM
中科院分区：
文献类型：
--
作者：
Akowuah, EF;Gray, C;Newman, CM
进行性大隐静脉移植物（SVG）狭窄和闭塞仍然是冠状动脉旁路移植术的主要限制，也是基因治疗的重要靶点。金属蛋白酶组织抑制剂3（TIMP-3）的离体腺病毒基因转移减少了不良的SVG重塑后的动脉化，但对在患者中使用病毒载体的担忧仍然存在。超声暴露（USE）在回声微泡（ECM）的存在下，大大增强了非病毒基因的传递。我们研究了超声增强的TIMP-3基因转染（UEGD）对猪SVG血管重构的影响。在50%（v/v）BR 14 ECM（Brdycin）的存在下，在1 MHz、1.8机械指数（MI）、6%占空比（DC）下使用USE在体外猪血管平滑肌细胞中实现最大荧光素酶活性（相对于单独裸质粒的3000倍）和TIMP-3转基因表达。因此，将这些条件用于随后的体内研究。约克郡白色猪接受未转染或已经历lacZ（对照）或TIMP-3质粒的离体UEGD的颈动脉插入SVG。在移植后28天，TIMP-3组的管腔和总血管面积显著更大（10.1 +/- 1.2和25.5 +/- 2.2 mm（2），与未转染（6.34 +/- 0.5和20.8 +/- 1.9 mm（2））或lacZ转染（6.1 +/- 0.7和19.7 +/- 1.2 mm（2））对照相比（P
Progressive saphenous vein graft (SVG) narrowing and occlusion remains a major limitation of coronary artery bypass grafting and is an important target for gene therapy. Ex vivo adenoviral gene transfer of tissue inhibitor of metalloproteinase 3 (TIMP-3) reduces adverse SVG remodelling postarterialization, but concerns remain over the use of viral vectors in patients. Ultrasound exposure ( USE) in the presence of echocontrast microbubbles (ECM) substantially enhances nonviral gene delivery. We investigated the effects of ultrasound-enhanced gene delivery (UEGD) of TIMP-3 plasmid on vascular remodelling in porcine SVG. Maximal luciferase activity (3000-fold versus naked plasmid alone) and TIMP-3 transgene expression in porcine vascular smooth muscle cells in vitro was achieved using USE at 1 MHz, 1.8 mechanical index (MI), 6% duty cycle ( DC) in the presence of 50% (v/v) BR14 ECM (Bracco). These conditions were therefore utilized for subsequent studies in vivo. Yorkshire White pigs received carotid interposition SVG that were untransfected or had undergone ex vivo UEGD of lacZ ( control) or TIMP-3 plasmids. At 28 d postgrafting, lumen and total vessel area were significantly greater in the TIMP-3 group (10.1 +/- 1.2 and 25.5 +/- 2.2 mm(2), respectively) compared to untransfected (6.34 +/- 0.5 and 20.8 +/- 1.9 mm(2)) or lacZ-transfected (6.1 +/- 0.7 and 19.7 +/- 1.2 mm(2)) controls (P