Development of topical gene delivery system for clinical application

临床应用局部基因递送系统的开发

基本信息

批准号：
15590133
负责人：
SASAKI Hitoshi
金额：
$ 2.24万
依托单位：
Nagasaki University Hospital of Medicine and Dentistry
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Gene therapy is a useful treatment for curing inborn and acquired diseases. The eye has a number of advantages as a target organ for gene delivery. The use of non-viral vectors attracts great interests for in vivo gene delivery because they lack some of the risks inherent in viral vector systemsWe have investigated the transfection activity of plasmid DNA (pDNA)/cationic liposome complexes to the eye in rabbits. Among the transfection experiments with various lipids, pDNA complexed with DOTMA/Chol liposomes showed the highest transfection activity after intravitreal injection. The complexes at a charge raio of +2.0 produced maximal gene expression in the eye. In conlusion, optimizing the lipid composition of cationic liposomes and the charge ratio of pDNA/cationic liposome complexes should enhance the transfection efficiency to the eye.We have examined the effect of blood components on gene delivery with polyethyleneimine (PEI) in a human endothelial cell line. The transfection activity with pDNA/PEI complex was markedly decreased in the presence of fetal bovine serum, bovine serum albumin, human blood, human plasma, cholesterol and fatty acids, although it was not affected by lecithin and bile salts. Ternary complexes using polyanion could decrease the influence of blood components on gene expression. On the other hand, the transfection activity with pDNA/PEI complex was increased 2-3 times by the presence of glucose. We also found that the gene expression of pDNA/PEI complex and pDNA/liposomes complex was influenced by the disease such as hepatitis.We have developed the ocular pharmacokinetic/pharmacodynamic (PK/PD) model of model drug, bunazosin, on the basis of the concentration-time profiles and hypotensive profiles of bunazosin. The PK/PD models can predict the concentrations of bunazosin in the ocular tissues and its hypotensive effect after instillation and ocular injection. This PK/PD model must be useful for designing the gene delivery system.

基因疗法是治愈天生和获得性疾病的有用疗法。眼睛具有许多优势作为基因递送的靶器官。非病毒载体的使用吸引了体内基因递送的极大兴趣，因为它们缺乏病毒载体系统中固有的一些风险，研究了质粒DNA（PDNA）/阳离子脂质体复合物的转染活性。在与各种脂质的转染实验中，与DOTMA/CHOL脂质体复合的pDNA在玻璃体内注射后的转染活性最高。 +2.0的电荷RAIO的复合物在眼中产生最大基因表达。结合性，优化阳离子脂质体的脂质组成以及pDNA/阳离子脂质体配合物的电荷比应提高眼睛的转染效率。我们已经检查了人类内皮细胞系中血液成分对基因递送（PEI）的基因递送的影响。在存在胎牛血清，牛血清白蛋白，人血，人血浆，胆固醇和脂肪酸的情况下，pDNA/PEI复合物的转染活性显着降低，尽管它不受卵磷脂和胆汁盐的影响。使用多支支着的三元复合物可以降低血液成分对基因表达的影响。另一方面，通过葡萄糖的存在增加了用pDNA/PEI复合物的转染活性2-3倍。我们还发现，PDNA/PEI复合物和pDNA/脂质体复合物的基因表达受到肝炎等疾病的影响。我们已经根据Bunazosin的模型药物（PK/PD）模型，基于Bunazosin的模型药物模型，基于Bunazosin的浓度和下降。 PK/PD模型可以预测眼部组织中Bunazosin的浓度及其滴注和眼注射后的降压作用。该PK/PD模型必须对设计基因输送系统有用。