Electroporation System for Cutaneous Gene Transfer

描述（由申请人提供）：本研究计划的长期目标是开发有效的体内基因递送系统。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递送到皮肤。这种系统的关键要素是再现性、便携性和对同一受试者进行多次应用的能力。体内电穿孔已被证明是增强质粒DNA递送的有效方式。皮肤是皮肤疾病、疫苗和几种代谢紊乱的基因治疗方案的有吸引力的靶点，因为它易于递送和监测。将电介导的递送成功递送到皮肤的关键考虑因素是用于施加电脉冲的施用器。施源器应治疗足够的区域，并具有柔性，以确保与组织良好接触。本研究旨在开发和证明组织适形电穿孔施加器可用于有效地递送质粒DNA。为实现这一目标，将实现以下三个具体目标：评估设计用于将质粒DNA递送至皮肤的新的适形电极阵列，建立用于最大表达的递送参数，并确定治疗的总面积与表达水平之间的关系; ii.基于特定目标i中建立的电参数，配置和测试包含用于质粒DNA递送的微针的适形皮肤施用器;以及iii.以确定在前两个特定目的中建立的系统是否可以递送编码适用于治疗目的的蛋白质的质粒，以确定最大表达水平的持续时间，并确定该时间是否可以通过进行多次递送程序来增加。该工作将在无毛豚鼠中进行。首先，将确定最佳电穿孔条件以使DNA递送和表达最大化。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法来将裸露的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.