Liu Jia Wei, Dunoyer-Geindre Sylvie, Blot-Chabaud Marcel, Sabatier Florence, Fish Richard J, Bounameaux Henri, Dignat-George Françoise, Kruithof Egbert K O
Division of Angiology and Hemostasis, Department of Internal Medicine, Faculty of Medicine and University Hospital of Geneva, Geneva, Switzerland.
J Vasc Res. 2010;47(2):157-67. doi: 10.1159/000250094. Epub 2009 Oct 22.
Inflammatory activation of the vascular endothelium is a major contributory factor to ischemic cardiovascular disease. Endothelial progenitor cells (EPCs) are being investigated for the treatment of ischemic disease or to coat vein grafts for bypass surgery. As an inflammatory environment might reduce their therapeutic efficacy, we sought to generate EPCs that are less sensitive to inflammatory activation. EPCs were obtained from human umbilical cord blood and transduced with a lentiviral vector for stable expression of A20, an anti-inflammatory protein. Nontransduced and green-fluorescent-protein-transduced cells were used as controls. Expression of A20 by EPCs did not modify cell morphology or expression of a panel of 20 proteins known to contribute to angiogenesis. Also, A20 had no effect on the capacity of EPCs to form tube-like structures in Matrigel. A20 expression reduced EPC activation by tumor necrosis factor-alpha and interleukin-1beta as determined from changes in vascular cell adhesion molecule 1 and E-selectin expression and decreased monocyte transmigration through a monolayer of EPCs. In conclusion, EPCs can be genetically modified to overexpress A20 in a stable fashion. These cells become less sensitive to inflammatory stimuli. This may be of interest in cell-based therapeutic approaches for clinical settings where inflammation is an important pathogenic factor.
血管内皮的炎症激活是缺血性心血管疾病的一个主要促成因素。内皮祖细胞（EPCs）正在被研究用于治疗缺血性疾病或为搭桥手术的静脉移植物进行涂层。由于炎症环境可能会降低它们的治疗效果，我们试图生成对炎症激活不太敏感的EPCs。EPCs从人脐带血中获取，并用慢病毒载体进行转导，以稳定表达抗炎蛋白A20。未转导的细胞和转导绿色荧光蛋白的细胞用作对照。EPCs中A20的表达并未改变细胞形态或一组已知有助于血管生成的20种蛋白质的表达。此外，A20对EPCs在基质胶中形成管状结构的能力没有影响。根据血管细胞黏附分子1和E-选择素表达的变化以及单核细胞通过EPCs单层的迁移减少来确定，A20的表达降低了肿瘤坏死因子-α和白细胞介素-1β对EPCs的激活作用。总之，EPCs可以通过基因改造以稳定的方式过表达A20。这些细胞对炎症刺激变得不太敏感。这对于炎症是重要致病因素的临床环境中基于细胞的治疗方法可能具有重要意义。
