Image-guided, intravascular hydrodynamic gene delivery to skeletal muscle in pigs.
Image-guided, intravascular hydrodynamic gene delivery to skeletal muscle in pigs.
复制标题
DOI：
10.1038/mt.2009.206
复制
发表时间：
2010-01
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
开发一种有效、安全、便捷的肌肉基因传递方法是实现肌肉相关疾病基因治疗的关键一步。为此，我们探索了将图像引导导管插入技术与流体动力输送原理相结合的可能性，以实现猪肌肉特异性基因转移。结果表明，基因转移效率与注射流量、注射压力和注射体积直接相关。在注射压力为300 psi、注射体积为体重的1.5%、注射流速为15 ml/sec的条件下，获得了最佳的基因递送效果。在此条件下，水动力注射含有pCMV-Luc（100µg/ml）的生理盐水，在水动力基因递送后5天，从目标肌肉中提取的蛋白质荧光素酶活性为106 -107相对光单位(RLU)/mg。免疫组化分析结果显示，后肢肌群转染效率为70-90%，转染细胞中报告基因持续表达2个月。除了暂时性水肿和肌酸磷酸激酶升高外，未观察到永久性组织损伤。这些结果表明，图像引导、静脉流体动力传递是一种有效、安全的骨骼肌基因传递方法。
Development of an effective, safe and convenient method for gene delivery to muscle is a critical step toward gene therapy for muscle-associated diseases. Toward this end, we have explored the possibility of combining the image-guided catheter insertion technique with the principle of hydrodynamic delivery to achieve muscle specific gene transfer in pigs. We demonstrate that gene transfer efficiency of the procedure is directly related to flow rate, injection pressure and injection volume. The optimal gene delivery was achieved at a flow rate of 15 ml/sec with injection pressure of 300 psi and injection volume equal to 1.5% of body weight. Under such a condition, hydrodynamic injection of saline containing pCMV-Luc (100 µg/ml) resulted in luciferase activity of 106 –107 relative light units (RLU)/mg of proteins extracted from the targeted muscle 5 days after hydrodynamic gene delivery. Result from immunohistochemical analysis revealed 70–90% transfection efficiency of muscle groups in the hind limb and persistent reporter gene expression for 2 months in transfected cells. With an exception of transient edema and elevation of creatine phosphokinase, no permanent tissue damage was observed. These results suggest that the image-guided, intravenous hydrodynamic delivery is an effective and safe method for gene delivery to skeletal muscle.
登录
查看更多内容
影响因子：
4.2
作者：
Zhang, GF;Budker, V;Wolff, JA
通讯作者：
Wolff, JA
影响因子：
12.4
作者：
Kamimura, Kenya;Suda, Takeshi;Liu, Dexi
通讯作者：
Liu, Dexi
影响因子：
12.4
作者：
Hagstrom, JE;Hegge, J;Wolff, JA
通讯作者：
Wolff, JA
影响因子：
5
作者：
HAWKINS, IF
通讯作者：
HAWKINS, IF
影响因子：
12.4
作者：
Hodges, BL;Taylor, KM;Scheule, RK
通讯作者：
Scheule, RK