Development of lipid particles targeted via sugar-lipid conjugates as novel nuclear gene delivery system
Development of lipid particles targeted via sugar-lipid conjugates as novel nuclear gene delivery system
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DOI：
10.1016/j.biomaterials.2007.09.039
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发表时间：
2008-02-01
期刊：
影响因子：
14
通讯作者：
Harashima, Hideyoshi
中科院分区：
文献类型：
--
作者：
Masuda, Tomoya;Akita, Hidetaka;Harashima, Hideyoshi
有效的核基因递送对于成功的基因治疗至关重要。这项研究开发了一种新颖的系统，通过多功能包膜型纳米器件（MEND）模拟腺病毒（Ad）进入核的机制。在此系统中，质粒 DNA (pDNA) 与聚阳离子缩合，然后封装在脂质膜中。为了将 MEND 靶向核孔复合体 (NPC)，在脂质包膜表面对糖作为 NPC 介导的核靶向装置进行了修饰。这是通过合成糖-胆固醇缀合物来实现的。 MEND 与 NPC 结合后，pDNA 核心随着脂质包膜的破裂而转移到细胞核中。与未修饰的 MEND 相比，糖修饰的 MEND 在非分裂和分裂细胞中表现出更高的转染效率。共聚焦显微镜证实，MEND 的糖修饰改善了 pDNA 的核转移。此外，脂质包膜的不稳定显着增强了转染活性：因此，核递送效率与脂质包膜稳定性密切相关。此外，通过实时PCR对细胞摄取和核转移过程的定量评估证实，表面糖影响核转移，但不影响细胞摄取。总之，通过使用糖修饰的 MEND 并优化脂质包膜稳定性，成功开发了一种用于 pDNA 核递送的新型系统。 (c) 2007 Elsevier Ltd. 保留所有权利。
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. (c) 2007 Elsevier Ltd. All rights reserved.