Electroporation System for Cutaneous Gene Transfer

描述(由申请人提供)：该研究计划的长期目标是开发高效的体内基因输送系统。该项目的目标是开发一种改进的电穿孔系统，用于将质粒DNA输送到皮肤。这种系统的关键要素是可重复性、便携性和对同一主题执行多个应用程序的能力。体内电穿孔已被证明是一种有效的促进质粒DNA传递的方法。皮肤是皮肤病、疫苗和几种代谢性疾病基因治疗方案的诱人靶点，因为它很容易输送和监测。成功地将电介质传递到皮肤的一个关键考虑因素是用于施加电脉冲的敷贴器。涂抹器应该处理足够的区域，并且要灵活，以确保与组织的良好接触。这项研究的目的是开发和证明一种组织整合的电穿孔涂布器可以用于有效地递送质粒DNA。为了实现这一目标，将完成以下三个具体目标：i.评估一种新的可整合的电极阵列，设计用于将质粒DNA输送到皮肤，建立最大表达的输送参数，并确定治疗总面积与表达水平之间的关系；根据在特定目标I和III中确定的电参数，配置和测试含有用于递送质粒DNA的微针的适配皮肤敷贴器；为了确定在前两个特定目的中建立的系统是否可以递送编码用于治疗目的的蛋白质的质粒，确定最大表达水平的持续时间，并确定是否可以通过执行多次递送程序来延长这一时间。这项工作将在无毛豚鼠身上进行。首先，将确定最优的电穿孔条件以最大限度地传递和表达DNA。这项工作还将建立处理区域的大小和表达水平之间的关系。这些信息将有助于建立一种有效的方式，将裸露的DNA运送到皮肤上。研究人员率先使用电穿孔进行体内给药，因此非常适合成功完成这项研究。

DESCRIPTION (provided by applicant): The long-range goal of this research program is the development of efficient in vivo gene delivery systems. The goal of this project is the development of an improved electroporation system for the delivery of plasmid DNA to the skin. Critical elements for such a system are reproducibility, portability and the ability to perform multiple applications to the same subject. In vivo electroporation has been shown to be an effective way of enhancing delivery of plasmid DNA. The skin is an attractive target for gene therapy protocols for cutaneous diseases, vaccines and several metabolic disorders because it is easily accessible for both delivery and monitoring. A critical consideration for successful delivery of electrically mediated delivery to the skin is the applicator used to apply the electric pulses. The applicator should treat a sufficient area and be flexible to assure good contact with the tissue. This study is designed to develop and to demonstrate that a tissue conformable electroporation applicator can be used to efficiently deliver plasmid DNA. To accomplish this goal the following three specific aims will be performed: i. to evaluate a new conformable electrode array designed for delivery of plasmid DNA to the skin, to establish delivery parameters for maximal expression and to determine the relationship between the total area treated and the level of expression; ii. to configure and test a conformable skin applicator containing microneedles for plasmid DNA delivery based on the electrical parameters established in Specific Aim i; and iii. to determine if the system established in the first two Specific Aims can deliver plasmids coding for proteins applicable for therapeutic purposes, to determine the duration of maximal expression levels and to determine if this time can be increased by performing multiple delivery procedures. The work will be performed in hairless guinea pigs. Initially, the optimal electroporation conditions to maximize DNA delivery and expression will be determined. This work will also establish the relationship between the size of the treated area and expression levels. This information will help establish an effective way to deliver naked DNA to the skin. The investigators have pioneered the use of electroporation for in vivo delivery so are well suited to successfully complete the study.