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

调节热量与阻抗谱相结合，改善和控制体内电穿孔 DNA 向皮肤的输送

• 批准号: 10375335
• 负责人: Mark J. Jaroszeski
• 金额: $ 36.78万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10375335
• 关键词: Address; Affect; Animal Model; Animals; Area; Biological; Cavia; Cell membrane; Cells; Characteristics; Clinical; Clinical Trials; Combined Modality Therapy; Complex; Custom; DNA; DNA delivery; Data; Devices; Dose; Electricity; Electrodes; Electroporation; Elements; Factor IX; Feedback; Frequencies; Gene Delivery; Genes; Goals; Heating; Hepatitis B Surface Antigens; Histologic; Image; Immune response; Individual; Investigation; Kinetics; Knowledge; Lead; Luciferases; Measurement; Measures; Mediating; Membrane Fluidity; Methods; Modeling; Nature; Outcome; Outcome Study; Patients; Persons; Physiologic pulse; Plasmids; Procedures; Process; Property; Public Health; Publishing; Reporter; Reproducibility; Research; Research Personnel; Serum; Skin; Skin Temperature; Spectrum Analysis; System; Technology; Temperature; Therapeutic; Time; Tissues; Variant; Work; base; clinical application; design; electric field; electric impedance; experimental study; gene therapy; human tissue; improved; in vivo; individual variation; physical process; plasmid DNA; preclinical study; primary outcome; response; success; temporal measurement; therapeutic DNA; therapeutic protein; transgene delivery; treatment site; uptake; voltage

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.

项目总结/摘要： 基因治疗尚未成为现实。一个障碍和普遍性是，体内递送基因的方法 还没有达到期望的可靠性和控制水平。体内电穿孔是一种用于 已经在临床前研究中获得了成功。这些都是使用 技术用于各种应用。总的来说，他们证明了该方法的物理基础使得它 适合任何组织。这些研究为100多项使用该技术的临床试验铺平了道路。 vivo.因此，这种DNA递送方法有明确的研究和临床应用。但是， 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摄取的电脉冲的特性对于特定的组织类型通常是固定的 基于动物模型的优化。这些可能对临床上的类似组织几乎没有可翻译性。 作为模型的设置可能与人体组织不同。此外，个体之间也存在差异， 单独的.因此，使用相同的电脉冲（或电剂量）将DNA递送到特定类型的细胞， 组织不可能每次在该组织类型中使用该方法时都是最佳的。不幸的是， 实时定制/适配电治疗的手段可以规避这一点。 问题并增加该方法的效率/可靠性。现有技术的另一个问题是， 电穿孔影响细胞膜，并且传统上在环境温度下进行。 适度升高的温度可能会影响结果，因为它们会影响膜流动性。这 所提出的R 01已经被设计为结合解决这两个方面以改善皮肤中的递送。 这项研究的基础是初步数据，这些数据表明， 单独使用定制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施动态而非固定的电穿孔脉冲递送 在皮肤治疗部位。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种电力 脉冲，测量脉冲之间的阻抗，并对电治疗进行调整，以控制 输送过程，同时保持用户设置的适度升高的温度。预计可靠性 并且通过实现更高的DNA递送/表达和减少的变异来增加控制。