Gene Delivery Agents for Ultrasound-Mediated Transfection

用于超声介导转染的基因递送剂

• 批准号: 7326481
• 负责人: Joshua J. Rychak
• 金额: $ 9.99万
• 依托单位: TARGESON, LLC
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7326481
• 关键词: Animal Disease Models; Animals; Antibodies; Biological Markers; Bioluminescence; Biotin; Caliber; Cell Adhesion Molecules; Charge; Chemistry; Clinical; Coupled; Coupling; Crohn' s disease; DNA; Development; Electroporation; Electrostatics; Equipment; Exhibits; Flow Cytometry; Focused Ultrasound Therapy; Frequencies; Gene Delivery; Gene Transfer; Genetic Materials; Goals; Hour; Image; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestines; Invasive; Lipids; Localized; Location; Luciferases; Measurement; Mediating; Methods; Microbubbles; Microbubbles Ultrasound Contrast Medium; Modality; Monoclonal Antibodies; Mucous Membrane; Mus; Peroxisome Proliferator-Activated Receptors; Plasmids; Population; Preparation; Procedures; Production; Range; Rattus; Recombinants; Reporter; Reporter Genes; Reproducibility; Research; Site; Streptavidin; Surface; Techniques; Testing; Tissues; Transfection; Translating; Ultrasonography; Variant; Viral; Viral Vector; Work; base; concept; density; desire; gel electrophoresis; gene therapy; in vivo; mouse model; mucosal addressin cell adhesion molecule-1; novel; optical imaging; plasmid DNA; pre-clinical research; shear stress; size; tool; zeta potential

DESCRIPTION (provided by applicant): We propose to develop a novel acoustically activated microbubble gene delivery vehicle for treatment of experimental inflammatory bowel disease. Microbubble ultrasound contrast agents composed of a cationic lipid shell are targeted to the adhesion molecule MAdCAM-1 by a monoclonal antibody immobilized to the surface of the agent. Plasmid DNA is coupled to the cationic surface of the agent via electrostatic interaction. Microbubbles are administered intravenously and accumulate at sites of intravascular MAdCAM-1 expression, corresponding to inflammatory foci. DNA is released from the agents at the target site by ultrasound-mediated microbubble destruction using a clinical ultrasound scanner. This method has shown efficacy for transfecting reporter genes to perivascular tissue in mice and rats. This strategy offers a safe, non-viral, non-invasive method of delivering genetic material in vivo. Selectivity is accomplished by targeting the agents to a molecular marker of inflammation, and ultrasound-mediated microbubble destruction is required for release of DNA from the agent surface. The stability of these gene delivery vehicles will be examined to assess the feasibility for commercial scale production of a research-grade tool for gene delivery. The ability of these agents to selectively transfect inflamed bowel will be examined using a luciferase reporter gene and bioluminescence imaging in a mouse model of inflammatory bowel disease. This project will result in the development of a novel and efficient gene delivery tool for pre-clinical research, which can eliminate the need for problematic viral vectors and invasive electroporation techniques. Additionally, this work has the potential to translate into a clinically viable method of treatment for inflammatory bowel disease. Gene therapy offers a treatment modality for Crohn's disease, for which only symptomatic treatment is currently available. Ultrasound-mediated microbubble gene delivery is a safe, non-viral, non-invasive and high- efficiency method of transfection. The development of targeted ultrasound-activated microbubble gene delivery vehicles presents a novel and effective method of accomplishing gene transfer in pre-clinical research. This technique will offer an alternative to viral and invasive electroporation transfection techniques, and offers a potential mode for accomplishing gene delivery in a clinical setting using existing ultrasound equipment.

描述（由申请人提供）：我们建议开发一种新型声激活微泡基因递送载体，用于治疗实验性炎症性肠病。由阳离子脂质壳组成的微泡超声造影剂通过固定在试剂表面的单克隆抗体靶向粘附分子 MAdCAM-1。质粒 DNA 通过静电相互作用与试剂的阳离子表面偶联。微泡通过静脉注射并积聚在血管内 MAdCAM-1 表达的部位，对应于炎症灶。使用临床超声波扫描仪通过超声波介导的微泡破坏，将 DNA 从靶位点的药剂中释放出来。该方法已显示出将报告基因转染至小鼠和大鼠血管周围组织的功效。该策略提供了一种安全、非病毒、非侵入性的体内传递遗传物质的方法。选择性是通过将药剂靶向炎症分子标记物来实现的，并且需要超声介导的微泡破坏来从药剂表面释放DNA。将检查这些基因递送载体的稳定性，以评估研究级基因递送工具商业规模生产的可行性。将使用荧光素酶报告基因和生物发光成像在炎症性肠病小鼠模型中检查这些试剂选择性转染炎症性肠的能力。该项目将开发一种用于临床前研究的新型高效基因传递工具，从而消除对有问题的病毒载体和侵入性电穿孔技术的需求。此外，这项工作有可能转化为临床上可行的炎症性肠病治疗方法。基因疗法为克罗恩病提供了一种治疗方式，目前只能对症治疗。超声介导的微泡基因递送是一种安全、非病毒、非侵入性、高效的转染方法。靶向超声激活微泡基因递送载体的开发为临床前研究中完成基因转移提供了一种新颖有效的方法。该技术将为病毒和侵入性电穿孔转染技术提供替代方案，并提供一种使用现有超声设备在临床环境中完成基因传递的潜在模式。

项目成果

Analysis of in vitro transfection by sonoporation using cationic and neutral microbubbles.

• DOI: 10.1016/j.ultrasmedbio.2010.05.014
• 发表时间: 2010-11
• 期刊: ULTRASOUND IN MEDICINE AND BIOLOGY
• 影响因子: 2.9
• 作者: Tlaxca, Jose L.;Anderson, Christopher R.;Klibanov, Alexander L.;Lowrey, Bryce;Hossack, John A.;Alexander, J. Steven;Lawrence, Michael B.;Rychak, Joshua J.
• 通讯作者: Rychak, Joshua J.