Department of Medical Engineering, Colleges of Medicine and Engineering, University of South Florida, Tampa, FL, USA.
Methods Mol Biol. 2021;2265:635-644. doi: 10.1007/978-1-0716-1205-7_43.
Gene electrotransfer (GET) is a reliable and effective physical method for in vivo delivery of plasmid DNA (pDNA). Several preclinical and clinical studies have utilized GET to deliver plasmids encoding immune stimulating genes for treatment of melanoma and other tumor types. Intratumor delivery of plasmids encoding cytokines directly to tumors can induce not only a local immune response, but a systemic one as well. To obtain an effective immune response, it is critical to achieve the appropriate expression pattern of the delivered transgene. Expression pattern (levels and kinetics) can be modified by manipulating the electrotransfer parameters. These parameters include the tissue target and the electric pulse parameters of pulse width, electric field, and pulse number. We have found that to induce a robust immune response, we needed only low to moderately elevated expression levels compared to controls. When developing a therapeutic protocol, it is important to establish what expression profile will enable the appropriate response. In this chapter we describe how to determine the appropriate GET protocol to achieve the expression profile that can result in the desired clinical response.
基因电转移(GET)是一种可靠且有效的物理方法,可将质粒 DNA(pDNA)体内递送至。一些临床前和临床研究已经利用 GET 来递送编码免疫刺激基因的质粒,用于治疗黑色素瘤和其他肿瘤类型。将编码细胞因子的质粒直接递送至肿瘤内,可以诱导不仅局部免疫反应,而且全身免疫反应。为了获得有效的免疫反应,关键是要实现所递送转基因的适当表达模式。可以通过操纵电转移参数来修饰表达模式(水平和动力学)。这些参数包括组织靶标和电脉冲参数,包括脉冲宽度、电场和脉冲数。我们发现,与对照相比,仅需低至中度升高的表达水平即可诱导出强大的免疫反应。在制定治疗方案时,重要的是要确定哪种表达谱能够产生适当的反应。在这一章中,我们描述了如何确定适当的 GET 方案以实现能够产生所需临床反应的表达谱。