Enhanced inhibition of neointimal hyperplasia by genetically engineered endothelial progenitor cells
Enhanced inhibition of neointimal hyperplasia by genetically engineered endothelial progenitor cells
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DOI：
10.1161/01.cir.0000121732.85572.6f
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发表时间：
2004-04-13
期刊：
影响因子：
37.8
通讯作者：
Dzau, VJ
中科院分区：
文献类型：
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作者：
Kong, D;Melo, LG;Dzau, VJ
背景-循环内皮祖细胞（EPCs）以前曾有报道。在这项研究中，我们研究了血管保护基因内皮一氧化氮合酶（eNOS）和血红素氧合酶-1（HO-1）在EPCs中的过表达增强其抑制新生内膜增生的能力的假设。方法和结果- EPCs从兔外周血中分离，在培养中扩增，并用表达人eNOS的假型逆转录病毒载体转导（eNOS-EPC）、HO-1（HO-1-EPC）或绿色荧光蛋白（GFP-EPC）。EPCs体外转导效率> 90%。对四组家兔（每组n = 5 - 6）进行颈总动脉球囊血管成形术。损伤后立即将约5 × 106个自体eNOS-EPC或HO-1-EPC移植到损伤血管中。对照动物接受等量的GFP-EPC或林格氏盐水。移植后2周，在转导的兔血管中检测到eNOS和HO-1转基因转录物和蛋白。内皮化增强EPC移植血管独立的基因转移。相对于生理盐水对照，GFP-EPC处理的血管中的新生内膜增厚显著减少。在用eNOS-EPCs处理的血管中，新生内膜尺寸进一步减小。令人惊讶的是，相对于GFP-EPCs，在用HO-1-EPCs处理的血管中没有观察到额外的减少。血栓形成发生在约50%的盐水处理的血管，但几乎没有在所有的EPC移植vessel.Conclusions -我们的结论是，移植自体内皮祖细胞过度表达eNOS在受伤的血管增强血管保护性能的重建内皮，导致抑制新生内膜增生。这种基于细胞的基因治疗策略可能对血管疾病的治疗有用。
Background - Circulating endothelial progenitor cells (EPCs) have been reported previously. In this study, we examined the hypothesis that overexpression of vasculoprotective gene endothelial nitric oxide synthase ( eNOS) and heme oxygenase-1 (HO-1) in EPCs enhances their ability to inhibit neointimal hyperplasia.Methods and Results - EPCs were isolated from rabbit peripheral blood, expanded in culture, and transduced with pseudotyped retroviral vectors expressing human eNOS (eNOS-EPCs), HO-1 (HO-1-EPCs), or green fluorescent protein (GFP-EPCs). Transduction efficiency of EPCs ex vivo was > 90%. Four groups of rabbits (n = 5 to 6 per group) were subjected to balloon angioplasty of the common carotid artery. Immediately after injury, approximate to 5 x 10(6) autologous eNOS-EPCs or HO-1-EPCs were transplanted into the injured vessel. Control animals received an equivalent number of GFP-EPCs or Ringer's saline. Two weeks after transplantation, eNOS and HO-1 transgene transcripts and proteins were detected in the transduced rabbit vessels. Endothelialization was enhanced in the EPC-transplanted vessels independently of gene transfer. Neointimal thickening was significantly reduced in the GFP-EPC-treated vessels relative to the saline control. Neointima size was further reduced in vessels treated with eNOS-EPCs. Surprisingly, no additional reduction was seen in vessels treated with HO-1-EPCs relative to GFP-EPCs. Thrombosis occurred in approximate to50% of the saline-treated vessels but was virtually absent in all EPC-transplanted vessels.Conclusions - We conclude that transplantation of autologous EPCs overexpressing eNOS in injured vessels enhances the vasculoprotective properties of the reconstituted endothelium, leading to inhibition of neointimal hyperplasia. This cell-based gene therapy strategy may be useful in treatment of vascular disease.