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

基本信息

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
项目类别：
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

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

展开

项目摘要

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.

该研究计划的远距离目标是开发有效的体内基因递送系统。该特定项目的目的是开发改进的微创系统以提供质粒DNA到皮肤。皮肤很容易获得，这使其成为基因疗法的绝佳目标应用是直接治疗皮肤疾病还是利用皮肤作为交付的仓库蛋白质直接进行全身治疗的循环。充分利用皮肤的易于访问性开发非接触方法是一种简单而直接的体内方法，以提供DNA和可以以微创的方式完成。我们一直在努力开发这种方法，以前开发了用于此输送的电转移的设备和协议。而这些设备已经有效地工作了，可以以相对无创的方式实现这一目标，仍然有必要在电极和组织目标之间接触。另外，实现所需的施加电压在某些情况下输送可能会导致细胞或组织损伤或潜在的不适。这是至关重要的开发一种替代方法，可以和电转移一样工作，但无需接触。一个其他考虑是开发一种方法和/或设备，以更好地控制交付和朝着更可预测和可再现的表达模式发展。为此，我们建议利用可以透化细胞并促进质粒的非热大气压力等离子体设备 DNA吸收。我们假设通过离子沉积在目标表面实现了传递组织，如果调节离子沉积水平，则可以控制表达水平。这要进一步开发和评估的新型等离子体设备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一个可以操作电池的小型便携式设备。这将是一个非接触式输送装置，将最小化或消除潜在的不适和/或细胞损伤。这以下特定目标将作为该项目的一部分进行。 1）评估非热气大气用于控制离子的血浆装置，可以沉积在组织表面上； 2）评估NTAP 产生各种离子以将质粒DNA递送到皮肤的设备，并确定持续时间最大表达水平，并通过执行多次交付来确定是否可以增加此时间程序； 3）确定在前两个目标中建立的系统是否可以传递编码的质粒治疗蛋白。调查人员在开发非热等离子体设备方面有广泛的经验和基因转移非常适合成功完成研究。