Image-Guided Hydrodynamic Gene Delivery

图像引导水动力基因传递

• 批准号: 7525862
• 负责人: DEXI LIU
• 金额: $ 41.69万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7525862
• 关键词: Animal Model; Animals; Area; Biochemistry; Biological Assay; CMV promoter; Catheters; Clinical; Clinical Trials; Complementary DNA; Computers; Data; Development; Devices; Disease; Drug or chemical Tissue Distribution; Effectiveness; Family suidae; Frequencies; Gene Delivery; Gene Expression; Gene Transfer; Genes; Genomics; Goals; Hemophilia B; Hepatocyte; Hereditary Disease; Human; Image; Imaging Techniques; Imaging technology; Immune response; Individual; Injection of therapeutic agent; Invasive; Laboratory Research; Liver; Lobe; Lobular; Long-Term Effects; Luciferases; Membrane; Methods; Mus; Nucleic Acids; Organ; Outcome; Permeability; Plasmids; Preparation; Procedures; Protein C Inhibitor; Public Health; Publishing; RNA; Range; Rodent; Rodent Model; Saline; Serum; Site; Structure; Tail; Techniques; Testing; Therapeutic; Time; Tissues; Toxic effect; Transfection; Transgenes; Translational Research; Veins; base; clinical application; concept; design; gene therapy; improved; intrahepatic; new technology; plasmid DNA; pressure; research study; transduction efficiency; transgene expression; vector

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.

描述（由申请人提供）：开发一种新的、改进的安全、位点特异性和高效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量的含有质粒 DNA 的盐水。我们在啮齿动物中证明，基于流体动力学的程序在基因递送至肝细胞方面具有优越性。该方法有效性的机制涉及肝脏血管系统中升高的流体动力压力，这增强了内皮通透性并在肝细胞中产生瞬时膜孔。利用这些早期发现，我们开发了一种计算机控制的注射装置，适用于人体流体动力学基因传递。在拟议的研究中，我们将把流体动力学基因递送与成熟的图像引导导管插入技术相结合，以猪作为动物模型来演示将位点特异性基因递送到肝细胞中。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将基因转移到各个肝叶的注射参数。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来说是最佳的。在具体目标2中，我们将采用顺序注射的方法来转染整个肝脏。逐叶方法旨在开发一种将最大基因转移到人类肝脏的程序，以治疗肝脏相关的遗传疾病。具体目标3旨在确定水动力基因转移对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的组织分布的长期影响。在具体目标 4 中，我们将检查重复基因转移的有效性。这项转化研究旨在验证新开发的注射装置并建立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。如果成功完成，拟议的研究将显着推进基因治疗领域的发展，并为基因传递提供新技术。 公共健康相关性：在拟议的研究中，我们将把流体动力学基因传递与成熟的图像引导导管插入技术相结合，以猪作为动物模型来演示将位点特异性基因传递到肝细胞中。拟议的研究旨在验证新开发的注射装置并建立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。如果成功完成，拟议的研究将显着推进基因传递领域并为基因治疗提供新技术。