肌肉注射编码血管内皮生长因子（VEGF）和乳糖酶基因（LacZ）的腺病毒载体后，对缺血兔后肢血管生成及副作用的评估

Evaluation of angiogenesis and side effects in ischemic rabbit hindlimbs after intramuscular injection of adenoviral vectors encoding VEGF and LacZ.

作者信息

Vajanto Ismo, Rissanen Tuomas T, Rutanen Juha, Hiltunen Mikko O, Tuomisto Tiina T, Arve Katja, Närvänen Outi, Manninen Hannu, Räsänen Heikki, Hippeläinen Mikko, Alhava Esko, Ylä-Herttuala Seppo

机构信息

A.I. Virtanen Institute, Kuopio University, Finland.

出版信息

J Gene Med. 2002 Jul-Aug;4(4):371-80. doi: 10.1002/jgm.287.

DOI:10.1002/jgm.287

PMID:12124979

Abstract

BACKGROUND

Recent studies have suggested the therapeutic potential of vascular endothelial growth factor (VEGF) gene therapy in ischemic skeletal muscle. However, only limited information is available about the effects of VEGF gene therapy in different regions of ischemic limbs, effects of control adenoviruses, and biodistribution of the transgenes after intramuscular (i.m.) administration. Here we studied angiogenesis and side effects of adenovirus-mediated VEGF and beta-galactosidase (LacZ) gene transfers in ischemic rabbit hindlimbs.

METHODS AND RESULTS

Ten days after induction of ischemia, rabbits were treated with i.m. injections of saline, LacZ adenovirus (AdLacZ; 2x10(10) pfu) or adenovirus encoding mouse VEGF(164) (AdVEGF; 2x10(10) pfu). In rabbits treated with AdVEGF an increase in serum VEGF(164) levels was detected by ELISA three and seven days after the gene transfer. 30 days after the gene transfer a positive effect on capillary density was observed in the thigh region both in rabbits treated with AdVEGF and AdLacZ compared with animals that received saline. On the other hand, AdVEGF and AdLacZ gene transfers had no effect on the capillary density in the calf region on day 30. A positive correlation between the capillary density and the number of collateral arteries was observed in the thigh. Hindlimb and testis edema and excess non-physiological growth of capillaries were detected as adverse effects of the AdVEGF gene therapy. Biodistribution analysis showed that the transgene was present not only in the target muscle, but also in ectopic tissues seven days after i.m. gene transfer.

CONCLUSIONS

The results suggest that a high dose of adenoviral vector encoding either AdVEGF or AdLacZ induces angiogenesis in the rabbit hindlimb ischemia model; i.m. injection of adenovirus leads to the transfection of ectopic organs; and AdVEGF gene transfer induces edema in ischemic skeletal muscle.

摘要

背景

近期研究提示血管内皮生长因子（VEGF）基因治疗在缺血性骨骼肌中有治疗潜力。然而，关于VEGF基因治疗在缺血肢体不同区域的效果、对照腺病毒的作用以及肌肉内（i.m.）给药后转基因的生物分布，仅有有限的信息。在此，我们研究了腺病毒介导的VEGF和β-半乳糖苷酶（LacZ）基因转移在缺血兔后肢中的血管生成及副作用。

方法与结果

缺血诱导10天后，兔接受i.m.注射生理盐水、LacZ腺病毒（AdLacZ；2×10¹⁰ pfu）或编码小鼠VEGF₁₆₄的腺病毒（AdVEGF；2×10¹⁰ pfu）。在接受AdVEGF治疗的兔中，基因转移后3天和7天通过ELISA检测到血清VEGF₁₆₄水平升高。基因转移30天后，与接受生理盐水的动物相比，接受AdVEGF和AdLacZ治疗的兔大腿区域的毛细血管密度有积极影响。另一方面，30天时AdVEGF和AdLacZ基因转移对小腿区域的毛细血管密度无影响。在大腿区域观察到毛细血管密度与侧支动脉数量之间呈正相关。检测到后肢和睾丸水肿以及毛细血管非生理性过度生长是AdVEGF基因治疗的不良反应。生物分布分析表明，i.m.基因转移7天后转基因不仅存在于靶肌肉中，还存在于异位组织中。