VEGF gene delivery to myocardium: deleterious effects of unregulated expression.
VEGF gene delivery to myocardium: deleterious effects of unregulated expression.
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DOI：
10.1161/01.cir.102.8.898
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发表时间：
2000-08
期刊：
影响因子：
37.8
通讯作者：
R. J. Lee;M. Springer;W. Blanco-Bose;R. Shaw;P. Ursell;H. Blau
中科院分区：
文献类型：
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作者：
R. J. Lee;M. Springer;W. Blanco-Bose;R. Shaw;P. Ursell;H. Blau
背景血管内皮生长因子（VEGF）正在研究用于缺血心肌的治疗性血管生成。已经测试了临时输送系统。本研究的目的是探讨通过使用成肌细胞介导的递送在心肌中持续表达VEGF的效果。方法和结果将表达鼠VEGF基因和来自逆转录病毒启动子的β-半乳糖苷酶（β-gal）基因的原代鼠成肌细胞（5 × 10（5）细胞溶于10 μ L含0.5%BSA的PBS中）通过皮下途径植入免疫缺陷小鼠（n=11）的心室壁。对照免疫缺陷小鼠（n=12）注射相同数量的仅表达β-gal基因的成肌细胞。在第14天至第16天之间，将存活的小鼠安乐死，并对心脏进行组织学处理。在实验组中，11只小鼠中有11只在第13天表现出无法茁壮成长; 5只死亡发生在第8天和第15天之间。对照组小鼠无并发症。组织化学证实6/6只存活的实验小鼠和12/12只对照小鼠成功植入成肌细胞（阳性β-gal反应产物）。组织学显示，在6只存活小鼠中，VEGF成肌细胞注射心肌中有6只出现类似血管瘤的壁内血管肿瘤。β-Gal表达细胞存在于血管肿瘤部位。免疫组化定位丰富的内皮型一氧化氮合酶和CD 31（血小板和内皮细胞粘附分子）的病变内，与内皮细胞的存在一致。结论：在该模型中，VEGF的不受调节的持续表达与（1）高生长失败率/死亡率和（2）植入部位内皮细胞源性壁内血管肿瘤的形成相关。这些结果强调了调节VEGF表达对于治疗性血管生成的重要性。
BACKGROUND Vascular endothelial growth factor (VEGF) is being investigated for therapeutic angiogenesis in ischemic myocardium. Primarily, transient delivery systems have been tested. The goal of this study was to investigate the effects of continuous expression of VEGF in myocardium by use of myoblast-mediated delivery. METHODS AND RESULTS Primary murine myoblasts (5 x 10(5) cells in 10 microL of PBS with 0.5% BSA) expressing both the murine VEGF gene and the beta-galactosidase (beta-gal) gene from a retroviral promoter were implanted in the ventricular wall of immunodeficient mice (n=11) via a subdiaphragmatic approach. Control immunodeficient mice (n=12) were injected with the same number of myoblasts expressing only the beta-gal gene. Between days 14 and 16, surviving mice were euthanized and the hearts processed for histology. In the experimental group, 11 of 11 mice demonstrated failure to thrive by day 13; 5 deaths occurred between days 8 and 15. There were no complications in the control mice. Histochemistry documented successful implantation of myoblasts (positive beta-gal reaction product) in 6 of 6 surviving experimental mice and 12 of 12 controls. Histology disclosed intramural vascular tumors resembling hemangiomas in the VEGF-myoblast-injected myocardium in 6 of 6 surviving mice. beta-Gal-expressing cells were present at the site of the vascular tumors. Immunohistochemistry localized abundant endothelial nitric oxide synthase and CD31 (platelet and endothelial cell adhesion molecule) within the lesion, consistent with the presence of endothelial cells. CONCLUSIONS In this model, unregulated continuous expression of VEGF is associated with (1) a high rate of failure to thrive/death and (2) formation of endothelial cell-derived intramural vascular tumors in the implantation site. These results underscore the importance of regulating VEGF expression for therapeutic angiogenesis.