Image-Guided Hydrodynamic Gene Delivery

描述（由申请人提供）：开发一种新的、改进的、安全的、位点特异性的、高效的基因传递方法已经成为人类基因治疗的主要焦点。为了加速实现这一目标，我们引入了水动力基因传递的概念。该过程需要快速尾静脉注射相对大体积的含质粒DNA的生理盐水。我们在啮齿类动物中证明了基于流体动力学的程序在基因传递到肝细胞方面是优越的。这种方法有效的机制涉及肝血管的动水压力升高，从而增强内皮细胞的通透性，并在肝细胞中产生短暂的膜孔。利用这些早期发现，我们开发了一种适用于人体流体动力基因传递的计算机控制注射装置。在拟议的研究中，我们将结合流体动力基因传递和一种成熟的图像引导导管插入技术，以猪为动物模型，演示特定部位的基因传递到肝脏肝细胞。本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特征。在具体目标1中，我们将建立程序并优化基因转移到肝脏各个叶的注射参数。我们将证实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转染整个肝脏。叶接叶的方法旨在开发一种最大限度地将基因转移到人类肝脏以治疗肝脏相关遗传疾病的程序。特异性目的3旨在确定水动力基因转移的长期影响，包括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反应以及转基因在组织中的分布。在具体目标4中，我们将检查重复基因转移的有效性。这项转化研究旨在验证一种新开发的注射装置，并建立一种适用于临床环境的程序。目的是收集准备临床试验所必需的数据。如果研究成功，将对基因治疗领域产生重大的推动作用，并为基因传递提供一种新的技术。

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.