Microbubble ultrasound improves the efficiency of gene transduction in skeletal muscle in vivo with reduced tissue damage
Microbubble ultrasound improves the efficiency of gene transduction in skeletal muscle in vivo with reduced tissue damage
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DOI：
10.1038/sj.gt.3301913
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发表时间：
2003-03-01
期刊：
影响因子：
5.1
通讯作者：
Blomley, MJK
中科院分区：
文献类型：
--
作者：
Lu, QL;Liang, HD;Blomley, MJK
肌肉注射裸质粒DNA是一种安全的系统传递治疗基因产品的方法，但效率有限。我们研究了使用微泡超声通过直接注射到活体小鼠骨骼肌来增加裸质粒DNA的输送，在幼鼠(4周)和老年(6个月)小鼠中都是如此。我们观察到，即使在没有超声的情况下，白蛋白包裹的微泡Optison(已获得患者的超声心动图许可)也显著提高了转染率。转基因表达的增加与年龄有关，因为Optison在老年小鼠中改善转基因表达的效率低于年轻小鼠。更重要的是，Optison显著减少了与裸质粒脱氧核糖核酸和阳离子聚合物PEI 25000的存在相关的肌肉损伤。中等功率的超声波(3W/cm(2)1 MHz，60 S照射，占空比20%)与Optison联合使用，可以提高老年小鼠的转染率，但对年轻小鼠没有影响。微泡和治疗性超声在临床上的安全使用，特别是微泡对组织损伤的保护作用，为体内肌肉基因传递提供了一种非常有前途的方法。
Intramuscular injection of naked plasmid DNA is a safe approach to the systemic delivery of therapeutic gene products, but with limited efficiency. We have investigated the use of microbubble ultrasound to augment naked plasmid DNA delivery by direct injection into mouse skeletal muscle in vivo, in both young (4 weeks) and older (6 months) mice. We observed that the albumin-coated microbubble, Optison (licensed for echocardiography in patients), significantly improves the transfection efficiency even in the absence of ultrasound. The increase in transgene expression is age related as Optison improves transgene expression less efficiently in older mice than in younger mice. More importantly, Optison markedly reduces muscle damage associated with naked plasmid DNA and the presence of cationic polymer PEI 25000. Ultrasound at moderate power (3 W/cm(2) 1 MHz, 60 s exposure, duty cycle 20%), combined with Optison, increases transfection efficiency in older, but not in young, mice. The safe clinical use of microbubbles and therapeutic ultrasound and, particularly, the protective effect of the microbubbles against tissue damage provide a highly promising approach for gene delivery in muscle in vivo.