Electroporation System for Cutaneous Gene Transfer

描述（由申请人提供）：本研究项目的长期目标是开发高效的体内基因传递系统。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输送到皮肤上。这种系统的关键要素是再现性、可移植性和对同一主题执行多个应用程序的能力。在体内电穿孔已被证明是一种有效的方式，提高质粒DNA的递送。对于皮肤病、疫苗和几种代谢紊乱的基因治疗方案来说，皮肤是一个有吸引力的靶标，因为它易于输送和监测。成功地将电介导的递送递送到皮肤的关键考虑因素是用于施加电脉冲的涂抹器。涂抹器应处理一个足够的区域，并灵活，以确保良好的接触组织。本研究旨在开发并证明组织适形电穿孔应用器可用于有效地传递质粒DNA。为了实现这一目标，将执行以下三个具体目标：1 .评估设计用于将质粒DNA传递到皮肤的新型合格电极阵列，建立最大表达的传递参数，并确定处理的总面积与表达水平之间的关系；2。配置和测试基于在Specific Aim i中建立的电参数的含有用于质粒DNA递送的微针的合格皮肤涂抹器；ⅲ。确定在前两个特定目标中建立的系统是否可以递送用于治疗目的的编码蛋白质的质粒，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个递送程序来增加这个时间。这项工作将在无毛豚鼠身上进行。首先，将确定最大化DNA传递和表达的最佳电穿孔条件。这项工作还将建立治疗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传递到皮肤上。研究人员率先使用电穿孔进行体内递送，因此非常适合成功完成研究。

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.