Controlled Delivery of Plasmid DNA via Low-Temperature Ion Deposition

通过低温离子沉积控制质粒 DNA 的传递

• 批准号: 9447306
• 负责人: RICHARD HELLER
• 金额: $ 42.47万
• 依托单位: OLD DOMINION UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9447306
• 关键词: Address; Air; Animals; Atmospheric Pressure; Blood Circulation; Cavia; Cells; Clinical Trials; DNA Vaccines; DNA delivery; Deposition; Development; Devices; Diagnostic; Dose; Electrodes; Electroporation; Erythropoietin; Gases; Gene Expression; Gene Targeting; Gene Transfer; Goals; In Vitro; Ions; Kinetics; Lead; Methods; Modeling; Nitrogen; Normal tissue morphology; Oncogenes; Oxygen; Pain; Pattern; Pharmacotherapy; Physiologic pulse; Plasma; Plasmids; Procedures; Production; Proteins; Protocols documentation; Reproducibility; Research; Research Personnel; Serum; Skin; Slide; Surface; System; Systemic Therapy; Techniques; Testing; Therapeutic; Time; Tissues; Ultrasonography; Variant; Work; Wound Healing; base; cold temperature; design; electric impedance; enzyme replacement therapy; experimental study; gene delivery system; gene therapy; improved; in vivo; minimally invasive; nanosecond; novel; plasmid DNA; portability; programs; public health relevance; skin disorder; success; therapeutic protein; three-dimensional modeling; uptake; vaccine delivery; voltage

The long-range goal of this research program is the development of efficient in vivo gene delivery systems. The goal of this specific project is the development of an improved minimally invasive system for the delivery of plasmid DNA to the skin. Skin is easily accessibility which makes it an excellent target for gene therapy applications whether it is for directly treating cutaneous diseases or utilizing the skin as a depot for delivering proteins directly to the circulation for systemic therapy. To take advantage of the easy accessibility of the skin it is critical to develop non-contact approaches that are a simple and direct in vivo method to deliver DNA and can be accomplished in a minimally invasive way. We have been working on developing such approaches and previously developed devices and protocols that utilized electrotransfer for this delivery. While these devices have worked effectively and can accomplish this in a relatively non-invasive manner, it is still necessary to have contact between the electrodes and the tissue target. In addition, the applied voltages needed to achieve delivery on some occasions may cause cellular or tissue damage or potential discomfort. It is critical to develop an alternative approach that can work as well as electrotransfer but do it without the contact. An additional consideration is to develop an approach and/or device that will allow for better control of delivery and move towards a more predictable and reproducible pattern of expression. To accomplish this, we propose to utilize a non-thermal atmospheric pressure plasma device that can permeabilize cells and facilitate plasmid DNA uptake. We hypothesize that delivery is achieved by ion deposition on the surface of the target tissue and that if the level of ion deposition is regulated then expression levels can be controlled. The novel plasma device to be further developed and evaluated is based on nanosecond pulsed air plasmas. Using this approach will allow us to develop a small portable device that could be battery operated. This will be a non-contact delivery device that will minimize or eliminate potential discomfort and/or cellular damage. The following specific aims will be performed as part of this project. 1) To evaluate non-thermal atmospheric plasma devices for controlled production of ions that can be deposited on tissue surface; 2) To evaluate NTAP devices producing various level of ions for delivery of plasmid DNA to the skin and to determine the duration of maximal expression levels and to determine if this time can be increased by performing multiple delivery procedures; and 3) to determine if the system established in the first two aims can deliver plasmids encoding therapeutic proteins. The investigators have extensive expereince in developing non-thermal plasma devices and gene transfer so are well suited to successfully complete the study.

该研究计划的长期目标是开发有效的体内基因传递系统。 这个特定项目的目标是开发一种改进的微创系统，用于提供 质粒 DNA 到皮肤。皮肤很容易接触，这使其成为基因治疗的绝佳目标 应用无论是直接治疗皮肤疾病还是利用皮肤作为输送的仓库 蛋白质直接进入循环系统进行全身治疗。利用皮肤的易接触性 开发非接触式方法至关重要，这种方法是一种简单而直接的体内方法来传递 DNA 和 可以通过微创方式完成。我们一直致力于开发此类方法并 之前开发的设备和协议利用电转移进行这种传递。虽然这些设备 已经有效地工作并且可以以相对非侵入性的方式实现这一点，但仍然有必要 电极和组织目标之间有接触。此外，实现所需的施加电压 在某些情况下分娩可能会导致细胞或组织损伤或潜在的不适。至关重要的是 开发一种替代方法，其效果与电转移一样好，但无需接触。一个 额外的考虑是开发一种方法和/或设备，以便更好地控制交付和 朝着更可预测和可重复的表达模式迈进。为了实现这一目标，我们建议 利用非热常压等离子体装置，可以透化细胞并促进质粒 DNA 摄取。我们假设传递是通过目标表面上的离子沉积来实现的 如果调节离子沉积水平，则可以控制表达水平。这 有待进一步开发和评估的新型等离子体装置基于纳秒脉冲空气等离子体。使用 这种方法将使我们能够开发出一种可以由电池供电的小型便携式设备。这将是一个 非接触式输送装置，可最大限度地减少或消除潜在的不适和/或细胞损伤。这 作为该项目的一部分，将实现以下具体目标。 1) 评估非热大气 用于受控产生可沉积在组织表面的离子的等离子体装置； 2）评估NTAP 产生不同水平离子的装置，用于将质粒 DNA 输送到皮肤并确定作用的持续时间 最大表达水平并确定是否可以通过执行多次递送来增加该时间 程序； 3) 确定前两个目标建立的系统是否可以传递编码质粒 治疗性蛋白质。研究人员在开发非热等离子体设备方面拥有丰富的经验 和基因转移因此非常适合成功完成研究。