Development of fusogenic liposomes which can deliver any substances into the cells through membrane fusion

开发融合脂质体，可通过膜融合将任何物质输送到细胞中

基本信息

批准号：
07557312
负责人：
MAYUMI Tadanori
金额：
$ 1.15万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
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-07557312/
关键词：
HVJ Liposomes Fusogenic liposomes Membrane fusion gene transfer Drug Delivery System

项目摘要

Many investigators have examined liposomes as carriers for transporting substances into cells. However, conventional liposomes do not directly fuse with the cell membrane. Even when they are actively bound to the cell surface by adding an antibody or ligand on the surface, they are taken into the cell by endocytosis. Therefore, most substances encapsulated in liposomes are degraded by lysosome enzymes, resulting in loss of their efficiency. On the other hand, the envelope of the Sendai virus contains a protein responsible for binding and fusion to the cell membrane, and Sendai virus genes are introduced into cells via membrane fusion. Therefore, the introduced genes are not decomposed by lysomome enzymes. Based on these findings, fusogenic liposomes which possess the envelope proteins of the Sendai virus on the liposomal surface were developed. Fusogenic liposomes can introduce their encapsulated materials into the cytoplasm through direct membrane fusion. In this study, we examined th … More e preparation and characterization of unilamellar fusogenic liposome, and the efficiency of delivery of proteins and DNA by unilamellar fusogenic liposomes.A fusogenic liposome was generated by fusion between a Sendai virus particle and a simple liposome (PA : PC : CHOL=1 : 4 : 5 (mol)). The liposomes with a negative surface charge of phosphatidic acid (PA) gave the best results. The amount of cholesterol (30-50%) and the presence of sialic acid did not affect the efficiency of delivery. In addition, we purified fusogenic liposomes based on the difference of the density of the particles. The purified fusogenic liposomes are similar in size and structure to the Sendai virus particle, and fuse with cells as efficiently as the Sendai virus. These liposomes can fuse with cells derived from various species (human, monkey, rabbit, canine, cattle, mouse and hamster) and from various tissues (fibroblasts, epithelial cells, endothelial cells, hepatocytes, neuronal cells and lymphoma cells) with almost the same efficiency. They can deliver any substances that can be encapsulated in the liposomes, including genes or proteins, into the cells.In conclusion, the fusogenic lipsomes are new tools for direct introduction of genetic information into animal tissues. Less

许多研究人员已经研究了脂质体作为将物质转运到细胞中的载体，但是，传统的脂质体不会直接与细胞膜融合，即使通过在表面添加抗体或配体将它们主动结合到细胞表面上，它们也会被带入细胞内。因此，大多数封装在脂质体中的物质都会被溶酶体酶降解，导致其效率丧失。另一方面，仙台病毒的包膜含有负责结合和融合的蛋白质。仙台病毒基因通过膜融合被引入细胞内，因此，引入的基因不会被溶酶体酶分解。基于这些发现，在脂质体表面具有仙台病毒包膜蛋白的融合脂质体。开发的融合脂质体可以通过直接膜融合将其封装材料引入细胞质中。在这项研究中，我们检查了单层融合脂质体的制备和表征。融合脂质体是通过仙台病毒颗粒和简单脂质体（PA：PC：CHOL=1：4：5（mol））之间的融合产生的。带有负表面电荷的磷脂酸（PA）给出了最好的结果，而胆固醇的量（30-50%）和唾液酸的存在则没有。此外，我们根据颗粒密度的差异纯化了融合脂质体，纯化的融合脂质体在大小和结构上与仙台病毒颗粒相似，并与仙台病毒一样有效地与细胞融合。这些脂质体可以与来自不同物种（人、猴、兔、犬、牛、小鼠和仓鼠）和不同组织（成纤维细胞、上皮细胞、内皮细胞、它们可以以几乎相同的效率将任何可以封装在脂质体中的物质（包括基因或蛋白质）递送到细胞中。总之，融合脂质体是直接引入基因的新工具。进入动物组织的信息较少。