Direct intramuscular gene transfer of naked DNA encoding vascular endothelial growth factor augments collateral development and tissue perfusion.
Direct intramuscular gene transfer of naked DNA encoding vascular endothelial growth factor augments collateral development and tissue perfusion.
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DOI：
10.1161/01.cir.94.12.3281
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发表时间：
1996-12
期刊：
影响因子：
37.8
通讯作者：
Y. Tsurumi;S. Takeshita;Dongfen Chen;M. Kearney;S. Rossow;J. Passeri;J. Horowitz;J. Symes;J. Isner
中科院分区：
文献类型：
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作者：
Y. Tsurumi;S. Takeshita;Dongfen Chen;M. Kearney;S. Rossow;J. Passeri;J. Horowitz;J. Symes;J. Isner
背景横纹肌已被证明能够吸收和表达以裸质粒DNA形式转移的外源基因，尽管通常具有低水平的基因表达。然而，在编码分泌蛋白的基因的情况下，低转染率可能不会排除分泌基因产物的生物活性。因此，我们研究了一种假说，即肌肉内注射编码血管内皮生长因子(VEGF)的裸质粒DNA可以增加后肢缺血动物模型的侧支发育和组织灌注量。方法与结果1只兔后肢缺血10d后，肌肉注射500微克phVEGF165或报告基因LacZ。30天后，与对照组相比，血管造影可识别的侧支血管和组织学可识别的毛细血管增加。这种增加的血管改善了对缺血肢体的血流灌注，由phVEGF165(0.85+/-0.05)比对照组(0.64+/-0.05，P<；.01)更高的小腿血压比率记录下来，改善了缺血肢体(用动脉内多普勒线测量)在静息时的血流量(phVEGF165=21.3+/-3.9毫升/分钟，对照=14.6+/-1.6毫升/分钟，P<；；.01)和血管扩张剂(phVEGF165=54.2+/-12.0mL/min，对照=37.3mL./-8.9mL/min，P<；.01)和增加内收肌中的微球(phVEGF165=4.3+/-1.6mL.min-1.100克组织-1，对照=2.9+/-1.2mL.min-1.100克组织-1，P<；.05)和腓肠肌(phVEGF165=3.90+/-1.0mL.min-1.100 g组织-1，对照组=2.8+/-1.4mL.min-1.100 g组织-1，P<0.05)。结论缺血骨骼肌是裸质粒DNA基因治疗的良好靶点。肌注血管生成细胞因子编码基因，特别是那些由完整细胞自然分泌的基因，可能成为广泛周围血管疾病患者的替代治疗策略，在这些患者中，基于血管内导管的基因转移的使用受到损害和/或禁止。
BACKGROUND Striated muscle has been shown to be capable of taking up and expressing foreign genes transferred in the form of naked plasmid DNA, although typically with a low level of gene expression. In the case of genes that encode secreted proteins, however, low transfection efficiency may not preclude bio-activity of the secreted gene product. Accordingly, we investigated the hypothesis that intramuscular (IM) gene therapy with naked plasmid DNA encoding vascular endothelial growth factor (VEGF) could augment collateral development and tissue perfusion in an animal model of hindlimb ischemia. METHODS AND RESULTS Ten days after ischemia was induced in one rabbit hindlimb, 500 micrograms of phVEGF165, or the reporter gene LacZ, was injected IM into the ischemic hindlimb muscles. Thirty days later, angiographically recognizable collateral vessels and histologically identifiable capillaries were increased in VEGF transfectants compared with controls. This augmented vascularity improved perfusion to the ischemic limb, documented by a superior calf blood pressure ratio for phVEGF165 (0.85 +/- 0.05) versus controls (0.64 +/- 0.05, P < .01), improved blood flow in the ischemic limb (measured with an intra-arterial Doppler wire) at rest (phVEGF165 = 21.3 +/- 3.9 mL/min, control = 14.6 +/- 1.6 mL/min, P < .01) and after a vasodilator (phVEGF165 = 54.2 +/- 12.0 mL/min, control = 37.3 +/- 8.9 mL/min, P < .01) and increased microspheres in the adductor (phVEGF165 = 4.3 +/- 1.6 mL.min-1.100 g of tissue-1, control = 2.9 +/- 1.2 mL.min-1.100 g of tissue-1, P < .05) and gastrocnemius (phVEGF165 = 3.9 +/- 1.0 mL.min-1.100 g of tissue-1, control = 2.8 +/- 1.4 mL.min-1.100 g of tissue-1, P < .05) muscles of the ischemic limb. CONCLUSIONS Ischemic skeletal muscle represents a promising target for gene therapy with naked plasmid DNA. IM transfection of genes encoding angiogenic cytokines, particularly those that are naturally secreted by intact cells, may constitute an alternative treatment strategy for patients with extensive peripheral vascular disease in whom the use of intravascular catheter-based gene transfer is compromised and/or prohibited.