In vivo liver electroporation:: Optimization and demonstration of therapeutic efficacy
In vivo liver electroporation:: Optimization and demonstration of therapeutic efficacy
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DOI：
10.1089/hum.2006.17.362
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发表时间：
2006-03-01
影响因子：
4.2
通讯作者：
Kon, OL
中科院分区：
文献类型：
--
作者：
Jaichandran, S;Yap, STB;Kon, OL
病毒载体介导的基因治疗的副作用（死亡和白血病发生）使质粒作为更安全、更可扩展、更简单和更具成本效益的载体重新受到关注。电穿孔和流体动力学递送是提高质粒介导的基因转移效率的两种技术。肝脏是用于靶向转移治疗相关基因以校正代谢紊乱（例如血友病A）的良好组织平台。然而，肝脏的体内电穿孔尚未显示出实现全身活性的分泌的转基因蛋白的治疗功效。我们研究了电场强度、脉冲持续时间、脉冲数、电波形、电极接触面积、质粒给药途径和注射技术对裸质粒体内电转移至肝脏效率的影响。质粒注射入体静脉上级肝内注射。与体内肌肉电穿孔不同，高压脉冲和微秒脉冲没有优势。最佳的电穿孔条件是8 - 10个20毫秒的单或双极脉冲，每个250 V/cm。使用非流体动力学技术，大大提高了电转移效率，最小的组织损伤，我们首次证明，在血友病小鼠中，肝脏定向的因子VIII cDNA的体内电穿孔实现了显着的表型校正。
Adverse effects (death and leukemogenesis) from viral vector-mediated gene therapy have renewed interest in plasmids as safer, more scalable, simple, and cost-effective vectors. Electroporation and hydrodynamic delivery are two techniques that improve the efficiency of plasmid-mediated gene transfer. The liver is a good tissue platform for targeted transfer of therapeutically relevant genes for correction of metabolic disorders, for example, hemophilia A. However, in vivo electroporation of liver has not yet been shown to achieve therapeutic efficacy of systemically active, secreted transgenic proteins. We have investigated the effect of field strength, pulse duration, pulse number, electrical waveforms, electrode contact area, plasmid administration routes, and injection technique on the efficiency of in vivo electrotransfer of naked plasmid to liver. Plasmid injection into a systemic vein was superior to intrahepatic injection. Unlike in vivo muscle electroporation, high-voltage pulses and microsecond pulses offered no advantage. Optimal electroporation conditions were 8 - 10 uni-or bipolar pulses of 20 msec, each at 250 V/cm. Using a nonhydrodynamic technique that greatly enhanced electrotransfer efficiency with minimal tissue injury, we demonstrate for the first time that liver-directed in vivo electroporation of factor VIII cDNA achieved significant phenotypic correction in hemophilic mice.