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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

我们开发了一种新的基因传递载体系统HVJ-脂质体。在该系统中，将DNA包埋在带负电荷的脂质体中，并将脂质体与紫外线灭活的仙台病毒HVJ融合，形成HVJ脂质体。HVJ-脂质体作为一种非病毒载体被广泛用于将DNA、寡核苷酸和蛋白质导入各种动物器官。利用该载体系统，我们成功地预防了大鼠球囊损伤动脉的再狭窄，抑制了肾小球硬化，保护了移植器官，并抑制了播散性脑肿瘤。然而，该系统仍需进一步改进，以获得更有效的体内、外给药系统，从而为将来的基因治疗奠定基础。当采用涡旋和挤压法制备含有阳离子DC-胆固醇（DC-Chol）的HVJ-阳离子脂质体时，FITC标记的反义寡核苷酸或核酶的给药能力大大增强，荧光素酶基因的表达也比常规方法提高了约70倍。 ...更多信息含有磷脂酰丝氨酸（PS）的天然HVJ-脂质体。HVJ-DC-Chol-脂质体可用于小鼠脑内播散性胶质母细胞瘤的自杀基因治疗。我们进一步优化脂质体的脂质组成，制备了一种新型的HVJ脂质体，其主要成分为鞘磷脂（Sph）、二油酰磷脂酰乙醇胺（DOPE）、磷脂酰胆碱（PC）、胆固醇（Chol）和DC-Chol（1.7：1.7：1.7：4.0：1.0摩尔比）作为在培养细胞中基因表达的最有效载体（比常规HVJ-脂质体高100-800倍）。然而，与常规HVJ脂质体相比，阳离子脂质体的使用反而降低了动物器官中的基因表达。然后，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）在肝脏或肌肉中的基因表达比常规HVJ-脂质体高5-10倍。因此，阳离子脂质体似乎在体外和体内基因表达中具有相互作用，并且我们的发现表明脂质体用于体外和体内基因转移的替代用途。少