Image-Guided Hydrodynamic Gene Delivery

描述（由申请人提供）：开发一种新的和改进的安全、位点特异性和有效的基因递送方法已成为人类基因治疗的主要焦点。为了加速实现这一目标，我们引入了流体动力学基因递送的概念。该过程需要快速尾静脉注射相对大体积的含有质粒DNA的盐水。我们在啮齿动物中证明，基于流体力学的程序在向肝细胞进行基因递送方面具有上级优势。该方法有效性的潜在机制涉及肝血管系统中的流体动力学压力升高，其增强内皮渗透性并在肝细胞中产生瞬时膜孔。利用这些早期的发现，我们已经开发了一个计算机控制的注射装置适用于流体动力学基因输送在人类。在本研究中，我们将结合联合收割机的流体动力学基因传递与一个发达的图像引导导管插入技术，以证明位点特异性基因传递到肝细胞作为动物模型的猪。本研究的目的是建立一种微创，易于重复的程序，能够实现转基因表达在假定的治疗水平，同时保持可接受的毒性概况。在具体目标1中，我们将建立程序并优化注射参数，以将基因转移到肝的各个叶。我们将确认在啮齿动物中建立的参数对于猪的肝脏基因转移是最佳的。在具体目标2中，我们将采用顺序注射方法来切除整个肝脏。该方法旨在开发一种最大限度地将基因转移到人类肝脏的方法，用于治疗肝脏相关遗传疾病。具体目标3旨在确定流体动力学基因转移在转基因表达持续性、对组织毒性的影响、对转基因产物的免疫应答和转基因的组织分布方面的长期影响。在具体目标4中，我们将检查重复基因转移的有效性。本转化研究旨在验证新开发的注射器械，并建立适用于临床环境的程序。目的是收集临床试验准备所必需的数据。如果成功完成，拟议的研究将大大推进基因治疗领域，并将提供一种新的基因递送技术。 公共卫生相关性：在本研究中，我们将结合联合收割机的流体动力学基因传递与一个发达的图像引导导管插入技术，以证明位点特异性基因传递到肝细胞作为动物模型的猪。拟定研究旨在确认新开发的注射器械，并建立适用于临床环境的程序。目的是收集临床试验准备所必需的数据。如果成功完成，拟议的研究将大大推进基因递送领域，并为基因治疗提供新技术。

DESCRIPTION (provided by applicant): Development of a new and improved method for safe, site specific, and efficient gene delivery has become a major focus for human gene therapy. To accelerate attaining this goal, we have introduced the concept of hydrodynamic gene delivery. The procedure entails a rapid tail vein injection of a relatively large volume of saline containing plasmid DNA. We demonstrated in rodents that the hydrodynamics-based procedure is superior in gene delivery to hepatocytes. The mechanism underlying the effectiveness of this method involves an elevated hydrodynamic pressure in the liver vasculature, which enhances endothelial permeability and generates transient membrane pores in hepatocytes. Using these earlier findings, we have developed a computer controlled injection device applicable to hydrodynamic gene delivery in human. In the proposed study, we will combine hydrodynamic gene delivery with a well-developed image-guided catheter insertion technique to demonstrate site-specific gene delivery into liver hepatocytes using swine as an animal model. The objective of this study is to establish a minimally invasive, easily repeatable procedure capable of achieving transgene expression at the putative therapeutic level while maintaining acceptable toxicity profiles. In specific aim 1, we will establish the procedure and optimize the injection parameters for gene transfer to individual lobes of the liver. We will confirm that parameters established in rodents are optimal for liver gene transfer in swine. In specific aim 2, we will take a sequential injection approach to transfect the entire liver. The lobe-by-lobe approach aims to develop a procedure for maximal gene transfer to human liver for treatment of liver related genetic diseases. Specific aim 3 is designed to determine the long-term effect of hydrodynamic gene transfer with respect to persistence of transgene expression, effect on tissue toxicity, immune response to transgene product, and tissue distribution of the transgene. In specific aim 4, we will examine the effectiveness of repeated gene transfer. This translational research is designed to validate a newly developed injection device and to establish a procedure applicable to a clinical setting. The objective is to collect data essential for preparation of clinical trials. If successfully accomplished, the proposed study will significantly advance the field of gene therapy and will provide a new technology for gene delivery. Public Health Relevance: In the proposed study, we will combine hydrodynamic gene delivery with a well-developed image-guided catheter insertion technique to demonstrate site-specific gene delivery into liver hepatocytes using swine as an animal model. The proposed study is designed to validate a newly developed injection device and to establish a procedure applicable to a clinical setting. The objective is to collect data essential for preparation of clinical trials. If successfully accomplished, the proposed study will significantly advance the field of gene delivery and provide a new technology for gene therapy.