Development of novel tissue directed ultra sound therapeutic gene delivery system

描述（由申请人提供）：低水平的高密度脂蛋白（HDL）胆固醇是冠心病（CHD）的重要危险因素。高密度脂蛋白胆固醇水平可预测他汀类药物治疗患者的主要心血管事件。在低密度脂蛋白胆固醇水平低于每分升70毫克的患者中也观察到这种关系。研究表明，多核苷酸，如DNA和RNA，可以被传递到细胞中，产生具有治疗作用的细胞变化。为了成功地实现基因治疗，至关重要的是，遗传物质与有效的递送载体一起工作，以提供局部治疗。编码表达全部或部分蛋白质的DNA既可以直接原位传递给生物体，也可以通过转移到然后移植到生物体中的细胞中间接传递。SonoGene公司正在研究哺乳动物的非病毒转染作为基因治疗的一种手段。我们提出了一种靶向转染DNA质粒到肝脏以提高apoA-I表达的方法。这项拨款的申请是为了支持研究开发一种合适的非侵入性途径，以诱导apoa - 1基因在肝脏中的表达，从而有利于改变胆固醇代谢。我们建议利用微球或微泡作为载体将裸质粒apoA-I DNA注射到血液中。肝脏的毛细血管内皮细胞是开孔的，可以增强质粒进入肝细胞的摄取。然后用超声波处理肝脏，破坏微球，导致肝内皮细胞短暂多孔，允许肝脏摄取和表达质粒DNA。公共卫生相关性：高密度脂蛋白（HDL）胆固醇水平是冠心病的重要危险因素。低高密度脂蛋白胆固醇水平可预测患者的主要心血管事件。SonoGene建议开发新的超声技术来提高高密度脂蛋白胆固醇水平，作为一种心脏保护治疗策略。

DESCRIPTION (provided by applicant): Low levels of high-density lipoprotein (HDL) cholesterol is an important risk factor for coronary heart disease (CHD). HDL cholesterol levels are predictive of major cardiovascular events in patients treated with statins. This relationship was also observed among patients with LDL cholesterol levels below 70 mg per deciliter. Studies have demonstrated that polynucleotides, such as DNA and RNA, can be delivered to cells producing cellular changes that can be therapeutic. In order to successfully achieve gene therapy, it is essential that the genetic materials work in conjunction with an effective delivery vehicle to provide localized therapy. DNA coded to express a whole or partial protein can be delivered either directly to the organism, in situ, or indirectly by transfer to a cell that is then transplanted into the organism. SonoGene Corporation is researching the non-viral transfection of mammals as a means of gene therapy. It is proposed to develop a method for targeting the transfection of DNA plasmid to the liver to enhance apoA-I expression. The request for this grant is to support research in developing a suitable non-invasive route to induce apoA-I gene expression in the liver for the purpose of favorably altering cholesterol metabolism. We propose to inject naked plasmid apoA-I DNA into the blood utilizing microspheres or microbubbles as a delivery vehicle. The capillary endothelial cells in the liver are fenestrated and permit enhanced uptake of plasmids into liver cells. The liver is then treated with ultrasound waves that disrupt the microspheres and cause transient porosity of liver endothelial cells, allowing for the uptake and expression of plasmid DNA by the liver. PUBLIC HEALTH RELEVANCE: High-density lipoprotein (HDL) cholesterol level is an important risk factor for coronary heart disease. Low HDL cholesterol levels are predictive of major cardiovascular events in patients. SonoGene proposes to develop novel ultrasound technology to raise HDL cholesterol levels as a cardio-protective therapeutic strategy.