Development of lipid particles targeted via sugar–lipid conjugates as novel nuclear gene delivery system
- 作者：Tomoya Masuda ; Hidetaka Akita ; Takashi Nishio ; Kenichi Niikura ; Kentaro Kogure ; Kuniharu Ijiro ; Hideyoshi Harashima
- 关键词：Non-viral vector ; Gene delivery ; Nuclear delivery ; Sugar ; Envelope-type nano device
- 刊名：Biomaterials
- 出版年：2008
- 期刊代码：7_01429612
- 类别：ms
- 出版时间：February 2008
- 卷：29
- 期：6
- 页码：709-723
- 文件大小：1146 K
摘要
Efficient nuclear gene delivery is essential for successful gene therapy. This study developed a novel system that mimics the mechanism of nuclear entry of adenovirus (Ad) by means of a Multifunctional Envelope-type Nano Device (MEND). In this system, plasmid DNA (pDNA) was condensed with polycation, followed by encapsulation in a lipid membrane. To target MEND to the nuclear pore complex (NPC), sugar served as a NPC-mediated nuclear targeting device was modified on the surface of the lipid envelope. This was accomplished via synthesis of a sugar–cholesterol conjugate. After binding of the MEND to the NPC, the pDNA core was transferred into the nucleus in conjunction with a breakdown of the lipid envelope. Sugar-modified MEND showed higher transfection efficiency compared with unmodified MEND, in non-dividing and dividing cells. Confocal microscopy confirmed that nuclear transfer of pDNA was improved by sugar modification of MEND. Furthermore, destabilization of the lipid envelope significantly enhanced transfection activity: therefore, nuclear-delivery efficiency was closely related to lipid envelope stability. Moreover, quantitative evaluation of cellular uptake and nuclear transfer processes by real-time PCR confirmed that the surface sugars affected nuclear transfer, but not cellular uptake. In summary, a novel system for the nuclear delivery of pDNA was successfully developed by using a sugar-modified MEND and by optimizing the lipid envelope stability.