Development of new DDS to individual organs by means of cell technology and its opplication to treatment of human diseases

利用细胞技术开发个体器官新型DDS及其在人类疾病治疗中的应用

基本信息

批准号：
07558126
负责人：
KANEDA Yasufumi
金额：
$ 6.21万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07558126/
关键词：
DDS (Drug Delivery System)Liposome HVJ-liposome Gene therapy 遺伝子導入

项目摘要

We developed a novel gene delivery vector system called HVJ-liposome. In this delivery system, DNA was entrapped into negative-charge liposome and the liposome was fused with UV-inactivated HVJ,Sendai virus, to form HVJ-liposome. HVJ-liposome is now widely used as an efficient non-viral vector for introducing DNA,oligonucleotides and proteins into various animal organs. Using this vector system, we succeeded in prevention of restenosis of balloon injured areteries of rats, suppression of glomerulosclerosis, protection of transplanted organs, and inhibition of disseminated brain tumors. However, the system should be improved for more efficient delivery system both in vitro and in vivo toward future gene therapy.When HVJ-cationic liposome having cationic DC-cholesterol (DC-Chol) was prepared by vortexing and extrusion, the delivery of FITC-labeled antisense oligonucleotides or ribozymes was greatly enhanced, and luciferase gene expression was also increased about 70 times more than conve … More ntional HVJ-liposome containing phosphatidylserine (PS). The HVJ-DC-Chol-liposome was very useful for suicide gene therapy of disseminated glioblastoma in mouse brain. We further optimized lipid components of liposome and developed a novel HVJ-liposome consisting of sphingomyelin (Sph), dioleoylphosphatidylethanolamine (DOPE), phosphatidylcholine (PC), cholesterol (Chol) and DC-Chol (1.7 : 1.7 : 1.7 : 4.0 : 1.0 in molar ratio) as the most efficient vector for gene expression in culture cells (100-800 times higher than conventional HVJ-liposome). However, the use of cationic lipids rather reduced gene expression in animal organs, compared with conventional HVJ-liposome. Then, HVJ-liposome containing Sph, DOPE,PC,PS and Chol (1.3 : 1.3 : 1.0 : 5.0 in molar ratio) mimicking HIV envelope (called HVJ-AVE liposome) raised gene expression in either liver or muscle 5-10 times more than conventional HVJ-liposome. Thus, cationic lipids appeared to have reciprocal effects in gene expression in vitro and in vivo, and our finding indicates the alternative usage of liposomes for in vitro and in vivo gene transfer. Less

我们开发了一种新型基因递送载体系统，称为HVJ-脂质体。在该递送系统中，DNA被包埋在负电荷脂质体中，并且脂质体与紫外线灭活的仙台病毒融合，形成HVJ-脂质体。现在广泛用作将DNA、寡核苷酸和蛋白质引入各种动物器官的有效非病毒载体，利用该载体系统，我们成功地预防了球囊损伤的再狭窄。然而，该系统还需要改进，以获得更有效的体外和体内递送系统，以实现未来的基因治疗。当HVJ-阳离子脂质体具有阳离子时。通过涡旋和挤压制备DC-胆固醇（DC-Chol），大大增强了FITC标记的反义寡核苷酸或核酶的递送，并且荧光素酶基因表达也比含有磷脂酰丝氨酸 (PS) 的传统 HVJ 脂质体增加了约 70 倍。HVJ-DC-Chol 脂质体对于小鼠脑中播散性胶质母细胞瘤的自杀基因治疗非常有用。脂质体的成分，并开发了一种由鞘磷脂（Sph）组成的新型HVJ脂质体，二油酰磷脂酰乙醇胺 (DOPE)、磷脂酰胆碱 (PC)、胆固醇 (Chol) 和 DC-Chol（摩尔比为 1.7 : 1.7 : 1.7 : 4.0 : 1.0）作为培养细胞中基因表达最有效的载体（比传统载体高 100-800 倍）传统的HVJ-脂质体）然而，阳离子脂质的使用相当降低了动物器官中的基因表达，与传统的HVJ-脂质体相比，含有Sph、DOPE、PC、PS和Chol（摩尔比为1.3：1.3：1.0：5.0）的模拟HIV包膜的HVJ-脂质体（称为HVJ-AVE脂质体）提高了任一肝脏中的基因表达。或肌肉比传统的 HVJ 脂质体多 5-10 倍因此，阳离子脂质似乎在体外和基因表达中具有相互影响。体内，我们的发现表明脂质体可以替代用于体外和体内基因转移。