Regulated Heat Combined with Impedance Spectroscopy to Improve and Control Electroporative DNA Delivery to the Skin in Vivo

项目摘要/摘要： 基因疗法还没有成为现实。一个障碍和概括是在体内传递基因的方法 尚未达到预期的可靠性和可控性水平。体内电穿孔是一种 提供在临床前研究中取得成功的DNA。这些操作是使用 适用于各种应用的技术。总而言之，他们证明了这种方法的物理基础使它 适用于任何组织。这些研究为100多项使用该技术的临床试验铺平了道路 活着。因此，这种DNA递送方法具有明确的研究和临床应用价值。但是，这种方法 可以改进，因为它仍然缺乏控制/可靠性。其中一个原因是 用于诱导DNA摄取的电脉冲的特征对于特定的组织类型通常是固定的 基于动物模型的最优化。这些在临床上对类似组织的可译性可能很小。 作为模型的设置可能与人体组织不同。此外，不同个体之间也存在差异。 个人的。因此，使用相同的电脉冲(或电剂量)将DNA传递到特定类型的 在该组织类型中使用该方法时，组织不太可能每次都是最佳的。不幸的是，这是 目前最先进的技术。实时定制/调整电处理的手段可以绕过这一点 发布并增加了该方法的效率/可靠性。目前技术水平的另一个问题是在活体内 电穿孔影响细胞膜，传统上是在环境温度下进行的。 适当升高温度可能会影响结果，因为它们会影响膜的流动性。这 建议的R01旨在结合这两个方面来解决这两个方面的问题，以改善经皮给药。 这项研究的基础是初步数据，这些数据表明，当 单独使用定制的脉冲或适度的温度升高，研究计划包括 在控制温度的同时实现动态而非固定的电穿孔脉冲传输 在皮肤治疗现场。该系统将利用组织电阻抗的实时测量 由电穿孔脉冲引起的变化。该系统将能够应用多个电力 脉冲，测量脉冲之间的阻抗，并对电气处理进行调整，以控制 送药程序，同时保持用户设定的适度高温。预计可靠性 通过实现更高的DNA传递/表达和减少变异，将增加控制。

PROJECT SUMMARY / ABSTRACT: Gene therapy is not yet a reality. One obstacle and generalization is that methods for delivering genes in vivo have not yet achieved a desired level of reliability and control. In vivo electroporation is a method for delivering DNA that has been successful in preclinical studies. These have been performed using the technology for a variety of applications. Collectively, they prove that the physical basis of the method makes it adaptable to any tissue. These studies paved the way for over 100 clinical trials that use the technology in vivo. Thus, there are clear research and clinical applications for this DNA delivery method. But, the method could be improved because it still suffers from lack of control/reliability. One reason for this is that the characteristics of the electric pulses used to induce DNA uptake are normally fixed for a particular tissue type based upon optimization in animal models. These may have little translatability to analogous tissues in clinical settings as models may not be identical to human tissues. In addition, there is variation from individual to individual. Thus, using the same electric pulses (or dose of electricity) to deliver DNA to a particular type of tissue is not likely to be optimal each time the method is used in that tissue type. Unfortunately, this is the current state of the art. A means of customizing/adapting electrical treatment in real-time could circumvent this issue and add to the efficiency/reliability of the method. Another issue with the state of the art is that in vivo electroporation affects cell membranes and has traditionally been performed at ambient temperature. Moderately increased temperatures could affect the results as they influence membrane fluidity. This proposed R01 has been designed to address these two aspects in combination to improve delivery in skin. The basis for this study is preliminary data that indicate approximately 10-fold increases in delivery when customized pulses or moderate temperature increases are used alone The research plan includes implementing electroporation pulse delivery that is dynamic rather than fixed while controlling the temperature at the skin treatment site. The system will utilize real-time measurements of tissue electrical impedance changes that result from electroporation pulses. The system will be capable of applying multiple electric pulses, measure impedance between pulses, and make adjustments to the electrical treatment to control the delivery procedure while maintaining a user set moderately elevated temperature. It is expected that reliability and control will be increased by achieving higher delivery/expression of the DNA and reduced variation.