Novel angiogenic gene therapy for the treatment of ischemic limb

在此之前，我们开发了一种新的血管生成疗法用于治疗缺血肢体，其中将经修饰的bFGF基因转导的自体成纤维细胞通过动脉内导管注入缺血肢体（离体方法）。在本研究中，我们确定了（1）多少基因转导的成纤维细胞诱导适当的治疗效果和（2）哪些血管生成生长因子有效地促进了侧支血管的发育1.离体基因治疗的适当控制我们将不同数量的基因（bFGF）转导的成纤维细胞（5x10^5个细胞、1x10^6个细胞、5x10^6个细胞和1x10^7个细胞）移植到兔后肢缺血模型，并在动脉内给药后28天评价血管生成作用。在用5 × 10^5个细胞或1 × 10^6个细胞处理的家兔中没有显著影响。在用5x10^6个细胞或1x10^7个细胞处理的动物中，观察到组织灌注显著改善，尽管在注射1x10^7个细胞后大量细胞分布到肺中。2. VEGF与bFGF将转染VEGF基因（VEGF组）或bFGF基因（bFGF组）的自体成纤维细胞以同样的方式注射到兔缺血肢体。治疗后28 d，bFGF组侧支传导和血管造影评分显著高于VEGF组，而bFGF组和VEGF组之间的血压比值和毛细血管密度无显著差异。这些数据表明，与VEGF相比，bFGF促进动脉生成。

Previously, we developed novel angiogenic therapy for the treatment of ischemic limbs, in which auto-fibroblasts, adenoviraIly transduced with modified bFGF gene, were infused into the ischemic limb thorugh an intra-arterial catheter (ex vivo method). In the present study, we determined (1)how many gene-transduced fibroblasts induced appropriate therapeutic effects and (2)which angiogenic growth factor effectively promoted the development of collateral vessels (VEGF vs bFGF).1.Appropriate control of the ex vivo gene therapyWe injected various numbers of gene (bFGF)-transduced fibroblasts (5x10^5 cells, 1x 10^6 cells, 5x10^6 cells and 1x10^7 cells) to the rabbit model of hind limb ischemia, and evaluated angiogenic effects at 28 days after intra-arterial administration. There were no significant effects in the rabbits treated with 5x10^5 cells nor 1x 10^6 cells. In the animals treated with 5x10^6 cells or 1x10^7 cells significant improvement of tissue perfusion was observed, though abundant cells were distributed to lung after injection of 1x10^7 cells. These findings indicated that 5x10^6 was appropriate number of cell administration.2.VEGF vs bFGFThe auto-fibroblasts, which were transferred with VEGF gene (VEGF group) or bFGF gene (bFGF group), were injected to rabbit ischemic limb in the same manner. Twenty-eight days after the treatment, collateral conductance and angiographic score in the bFGF group was significantly higher than those in the VEGF group, while no significant difference was detected in calf blood pressure ratio and capillary density between the bFGF and VEGF groups. These data suggested that bFGF promoted arteriogenesis as compared with VEGF.