Appropriate control of ex vivo gene therapy delivering basic fibroblast growth factor promotes successful and safe development of collateral vessels in rabbit model of hind limb ischemia
Appropriate control of ex vivo gene therapy delivering basic fibroblast growth factor promotes successful and safe development of collateral vessels in rabbit model of hind limb ischemia
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DOI：
10.1016/j.jvs.2003.09.016
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发表时间：
2004-03-01
影响因子：
4.3
通讯作者：
Shigernatsu, H
中科院分区：
文献类型：
--
作者：
Ishii, S;Koyama, H;Shigernatsu, H
目的：在我们先前的研究中，腺病毒介导的碱性成纤维细胞生长因子的离体基因转移促进了兔后肢缺血模型中显著的侧支血管发育。本研究探讨了如何控制这种基因治疗的有效性和安全性，并评估了重复应用该程序的可行性。方法：将带有分泌信号序列的修饰的hFGP基因通过腺病毒转移到培养的自体成纤维细胞中，并将不同数量的细胞在21天前切除左股动脉的家兔中，通过左髂内动脉注射（2 × 10（5）、1 × 10（6）、5 × 10（6）或2.5 × 10（7））或溶媒。细胞给药后28天，测量小牛血压比、血管造影评分、髂内动脉血流量和肌肉组织毛细血管密度，以分析缺血肢体中的侧支血管发育和组织灌注。为了评估递送效率和病毒污染，在用不同数量的基因转导细胞处理的兔中检查注射细胞的分布和血液抗腺病毒抗体滴度的时间过程。此外，动物接受两次注射，间隔21天，用含有荧光素酶基因的腺病毒载体感染的成纤维细胞，并测量荧光素酶表达以评估本发明的治疗是否可重复。在细胞施用后28天，在接受5 × 10（6）或2.5 × 10（7）个细胞的兔子中观察到显著的侧支血管发育而没有可检测的副作用，与接受赋形剂的动物相比，在细胞少于5 × 10（6）的动物中未检测到显著的发育（P
Purpose: In our previous study, adenovirus -mediated ex vivo gene transfer of basic fibroblast growth factor promoted significant collateral vessel development in a rabbit model of hind limb ischemia. The present study examined how to control the efficacy and safety of this gene therapy, and also evaluated the feasibility of repeat application of this procedure.Methods: Modified hFGP gene with the secretory signal sequence was adenovirally transferred to cultured autologous fibroblasts, and various numbers of the cells (2 X 10(5),1 X 10(6),5 X 10(6), or 2.5 X 10(7)) or vehicle was injected through the left internal iliac artery in rabbits in whom the left femoral artery had been excised 21 days previously. Twenty-eight days after cell administration, calf blood pressure ratio, angiographic score, blood flow in the internal iliac artery, and capillary density of muscle tissue were measured to analyze collateral vessel development and tissue perfusion in the ischemic limb. To assess delivery efficiency and viral contamination, the distribution of injected cells and the time course of blood anti-adenovirus antibody titer were examined in rabbits treated with various numbers of gene-transduced cells. In addition, animals received two injections, 21 days apart, of fibroblasts infected with adenovirus vector containing the luciferase gene, and luciferase expression was measured to evaluate whether the present therapy is repeatable.Results: At 28 days after cell administration, significant collateral vessel development without detectable side effects was observed in rabbits who received 5 X 10(6) or 2.5 X 10(7) cells, compared with those who received vehicle, and no significant development was detected in animals with fewer than 5 X 10(6) cells (P