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。HDL胆固醇水平是他汀类药物治疗患者主要心血管事件的预测指标。在LDL胆固醇水平低于70 mg/dl的患者中也观察到这种关系。研究已经证明，多核苷酸，如DNA和RNA，可以被递送到细胞，产生可以是治疗性的细胞变化。为了成功地实现基因治疗，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可以直接原位递送至生物体，或通过转移至细胞然后移植至生物体中而间接递送。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。本研究拟建立一种靶向转染DNA质粒至肝脏以增强apoA-I表达的方法。该基金的申请是为了支持开发合适的非侵入性途径的研究，以诱导apoA-I基因在肝脏中表达，从而有利地改变胆固醇代谢。我们建议利用微球或微泡作为递送载体将裸质粒apoA-I DNA注射到血液中。肝脏中的毛细血管内皮细胞是有孔的，允许质粒被增强地摄取到肝细胞中。然后用超声波处理肝脏，超声波破坏微球并引起肝脏内皮细胞的瞬时多孔性，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HDL胆固醇水平可预测患者的主要心血管事件。SonoGene提出开发新的超声技术，以提高HDL胆固醇水平作为心脏保护治疗策略。

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.