Image-Guided Hydrodynamic Gene Delivery

描述(由申请人提供)：开发一种新的、改进的安全、部位特异和有效的基因传递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基因传递的概念。这一过程需要快速尾静脉注射相对大容量的含有质粒DNA的生理盐水。我们在啮齿类动物身上证明了基于流体动力学的程序在基因传递方面优于肝细胞。这种方法有效的机制涉及到肝血管内流体动力压力的增加，这增加了内皮的通透性，并在肝细胞中产生了暂时性的膜孔。利用这些早期的发现，我们已经开发出一种计算机控制的注射装置，适用于人类的流体动力基因输送。在这项拟议的研究中，我们将结合流体动力基因传递和成熟的图像引导导管插入技术，以猪为动物模型来演示位置特异性基因传递到肝细胞中。这项研究的目的是建立一种微创、易于重复的程序，能够在假定的治疗水平上实现转基因表达，同时保持可接受的毒性分布。在具体目标1中，我们将建立基因转移到肝脏各个叶的程序和优化注射参数。我们将确认在啮齿类动物身上建立的参数是猪肝脏基因转移的最佳参数。在特定的目标2中，我们将采取序贯注射的方法来转染整个肝脏。这种逐叶转移的方法旨在开发一种最大限度地将基因转移到人类肝脏的程序，用于治疗与肝脏相关的遗传病。具体目的3旨在确定流体动力基因转移在转基因表达的持久性、对组织毒性的影响、对转基因产物的免疫应答以及转基因的组织分布方面的长期效应。在具体目标4中，我们将检查重复基因转移的有效性。这项转化性研究旨在验证新开发的注射设备，并建立适用于临床环境的程序。其目的是收集临床试验准备所必需的数据。如果成功完成，这项研究将大大推进基因治疗领域的发展，并将为基因输送提供一种新的技术。 公共卫生相关性：在拟议的研究中，我们将把流体动力基因传递与成熟的图像引导导管插入技术相结合，以猪为动物模型来演示位置特异性基因传递到肝细胞中。这项拟议的研究旨在验证一种新开发的注射装置，并建立适用于临床环境的程序。其目的是收集临床试验准备所必需的数据。如果成功完成，这项研究将极大地促进基因传递领域的发展，并为基因治疗提供一种新技术。

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.