Center for Experimental Drug and Gene Electrotransfer, Department of Oncology 54B1, Copenhagen University Hospital Herlev, 75 Herlev Ringvej, DK-2730 Herlev, Denmark.
Bioelectrochemistry. 2011 Oct;82(2):95-102. doi: 10.1016/j.bioelechem.2011.06.001. Epub 2011 Jun 12.
Gene electrotransfer is a non-viral technique using electroporation for gene transfection. The method is widely used in the preclinical setting and results from the first clinical study in tumours have been published. However, the preclinical studies, which form the basis for the clinical trials, have mainly been performed in rodents and the body of evidence on electrode choice and optimal pulsing conditions is limited. We therefore tested plate and needle electrodes in vivo in porcine skin, which resembles human skin in structure. The luciferase (pCMV-Luc) gene was injected intradermally and subsequently electroporated. Simultaneously, studies with gene electrotransfer to porcine skin using plasmids coding for green fluorescent protein (GFP) and betagalactosidase were performed. Interestingly, we found needle electrodes to be more efficient than plate electrodes (p<0.001) and electric field calculations showed that penetration of the stratum corneum led to much more homogenous field distribution at the DNA injection site. Furthermore, we have optimised the electric pulse regimens for both plate and needle electrodes using a range of high voltage and low voltage pulse combinations. In conclusion, our data support that needle electrodes should be used in human clinical studies of gene electrotransfer to skin for improved expression.
基因电转移是一种使用电穿孔进行基因转染的非病毒技术。该方法在临床前研究中得到广泛应用，并且已经发表了来自肿瘤的第一个临床研究结果。然而，作为临床试验基础的临床前研究主要在啮齿动物中进行，关于电极选择和最佳脉冲条件的证据有限。因此，我们在猪皮中进行了体内板状和针状电极测试，猪皮在结构上类似于人皮。将荧光素酶（pCMV-Luc）基因皮内注射，然后进行电穿孔。同时，还进行了使用编码绿色荧光蛋白（GFP）和β-半乳糖苷酶的质粒对猪皮进行基因电转移的研究。有趣的是，我们发现针状电极比板状电极更有效（p<0.001），并且电场计算表明，角质层的穿透导致在 DNA 注射部位的电场分布更加均匀。此外，我们使用一系列高压和低压脉冲组合优化了板状和针状电极的电脉冲方案。总之，我们的数据支持在皮肤基因电转移的人体临床研究中使用针状电极以提高表达。
