Electric Field - Forced Gene Transfer In Solid Tumors

电场 - 实体瘤中的强制基因转移

• 批准号: 6475237
• 负责人: FAN YUAN
• 金额: $ 32.58万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-11 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6475237
• 关键词: apoptosis; athymic mouse; biotechnology; cell line; collagen; electric field; electroporation; endopeptidases; experimental designs; extracellular matrix; gene delivery system; gene therapy; interleukin 12; intracellular transport; method development; mucopolysaccharides; neoplasm /cancer therapy; plasmids; skin transplantation; transfection; transposon /insertion element

Description (provided by applicant): The overall objective of this application is to improve electric field-forced gene transfer in solid tumors. Gene therapy has a great potential to improve treatment of different diseases, including solid tumors, but it has been impeded by the difficulties of gene delivery to specific cells. Recent reports by other investigators and ourselves have demonstrated that plasmid DNA delivery can be enhanced through the use of pulsed electric fields. However, present methods for optimizing electric field-forced gene transfer, in terms of electric pulse characteristics (e.g., field strength and pulse duration), are still empirical and incomplete. The fundamental hypothesis in this proposal is that electric field-forced DNA transport in the vicinity of cells determines the amount of gene transfer through the transient pores in the plasma membrane of cells created during in vivo electroporation. To test this hypothesis, we will systematically quantify the mechanisms of interstitial transport of DNA in response to different electric pulses both ex vivo (Aim 1) and in vivo (Aim 2). The quantification will be based on unique experimental methods developed in our labs, and it may result in an identification of effective pulse sequences for improving interstitial transport of DNA. The improved transport in the identified electric fields will be further increased through modifications in tumor tissue structures, using matrix enzymes, hypertonic solutions, or apoptotic agents (Aim 3). These chemical treatments transiently increase the interstitial space and thus decrease the resistance to DNA transport. Quantitative results from the transport studies in Specific Aims 1 through 3 will finally be used to improve transfection efficiency and therapeutic efficacy of interleukin- 12 gene in solid tumors transplanted in mice (Aim 4). The goal of the proposed study is to establish effective experimental protocols for improving interstitial transport of plasmid DNA which in turn may result in an increase in transfection efficiency in solid tumors without increasing toxicity.

描述（由申请人提供）：本申请的总体目标 是为了改善实体瘤中的电场造成基因转移。基因疗法 具有改善不同疾病治疗的巨大潜力，包括 实体瘤，但由于基因递送的困难而阻碍了 特定细胞。其他调查人员和我们自己的最新报告 证明可以通过使用来增强质粒DNA的递送 脉冲电场。但是，目前优化电动的方法 在电脉冲特性方面（例如， 野外强度和脉搏持续时间），仍然是经验和不完整的。这 该提案中的基本假设是电场效力的DNA 细胞附近的运输决定基因转移的量 通过在IN期间产生的细胞的质膜中的瞬态孔 体内电穿孔。为了检验这一假设，我们将系统地量化 DNA响应不同的DNA的间隙运输机制 电脉冲均离体（AIM 1）和体内（AIM 2）。定量 将基于我们实验室中开发的独特实验方法，它可能 导致鉴定有效脉冲序列以改进 DNA的间质运输。确定的运输改善 通过修饰肿瘤组织将进一步增加电场 结构，使用基质酶，高渗溶液或凋亡剂 （目标3）。这些化学处理瞬时增加了间隙空间 从而降低对DNA转运的抗性。定量结果 特定目标1到3的运输研究最终将用于 提高白介素-12的转染效率和治疗功效 在小鼠中移植的实体瘤中的基因（AIM 4）。提议的目标 研究是为了建立有效的实验方案以改善 质粒DNA的间质转运又可能导致增加 在实体瘤的转染效率方面，没有增加毒性。