Efficiency of High- and Low-Voltage Pulse Combinations for Gene Electrotransfer in Muscle, Liver, Tumor, and Skin
Efficiency of High- and Low-Voltage Pulse Combinations for Gene Electrotransfer in Muscle, Liver, Tumor, and Skin
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DOI：
10.1089/hum.2008.060
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发表时间：
2008-11-01
影响因子：
4.2
通讯作者：
Mir, L. M.
中科院分区：
文献类型：
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作者：
Andre, F. M.;Gehl, J.;Mir, L. M.
基因电转移作为一种有效的非病毒基因转移方法，正在获得越来越多的发展势头。在骨骼肌中，数据表明，电脉冲起着两个作用：在结构上透化肌纤维，并在药理学上支持DNA朝向或穿过透化膜的迁移。为了进一步研究这一点，在啮齿动物模型的肌肉、肝脏、肿瘤和皮肤中研究了透化短高压脉冲（HV;数百V/cm）和主要电泳长低压脉冲（LV;数十V/cm）的组合。进行了以下观察：（1）发现各种组织之间存在显著差异，可能与细胞大小和组织结构有关;（2）如果HV脉冲和LV脉冲之间存在时间间隔，基因表达增加;（3）HV脉冲是肌肉、肿瘤和皮肤的高电转移所需的，但不是肝脏;和（4）用低于透化的可检测阈值的HV场强度实现有效的基因电转移;和（5）HV和LV脉冲之间的滞后时间间隔降低了对HV脉冲的敏感性，使得能够实现更宽的HV幅度范围。总之，HV加LV脉冲代表了未来临床试验的有效和安全的选择，我们建议将基因转移到各种类型的组织中。
Gene electrotransfer is gaining momentum as an efficient methodology for nonviral gene transfer. In skeletal muscle, data suggest that electric pulses play two roles: structurally permeabilizing the muscle fibers and electrophoretically supporting the migration of DNA toward or across the permeabilized membrane. To investigate this further, combinations of permeabilizing short high-voltage pulses (HV; hundreds of V/cm) and mainly electrophoretic long low-voltage pulses (LV; tens of V/cm) were investigated in muscle, liver, tumor, and skin in rodent models. The following observations were made: (1) Striking differences between the various tissues were found, likely related to cell size and tissue organization; (2) gene expression is increased, if there was a time interval between the HV pulse and the LV pulse; (3) the HV pulse was required for high electrotransfer to muscle, tumor, and skin, but not to liver; and (4) efficient gene electrotransfer was achieved with HV field strengths below the detectability thresholds for permeabilization; and (5) the lag time interval between the HV and LV pulses decreased sensitivity to the HV pulses, enabling a wider HV amplitude range. In conclusion, HV plus LV pulses represent an efficient and safe option for future clinical trials and we suggest recommendations for gene transfer to various types of tissues.