Jiang Qian, Yue Dong, Nie Yu, Xu Xianghui, He Yiyan, Zhang Shiyong, Wagner Ernst, Gu Zhongwei
National Engineering Research Center for Biomaterials, Sichuan University , Chengdu 610064, Sichuan, P. R. China.
Center for Drug Research, Department of Pharmacy, Pharmaceutical Biology-Biotechnology, and Center for NanoScience (CeNS), Ludwig-Maximilians-Universitat , Butenandtstrasse 5-13, D-81377, Munich, Germany.
Mol Pharm. 2016 Jun 6;13(6):1809-21. doi: 10.1021/acs.molpharmaceut.5b00967. Epub 2016 May 12.
Cationic lipid based assemblies provide a promising platform for effective gene condensation into nanosized particles, and the peripheral properties of the assemblies are vital for complexation and interaction with physical barriers. Here, we report three cationic twin head lipids, and each of them contains a dioleoyl-glutamate hydrophobic tail and a twin polar head of lysine, arginine, or histidine. Such lipids were proven to self-assemble in aqueous solution with well-defined nanostructures and residual amino-, guanidine-, or imidazole-rich periphery, showing strong buffering capacity and good liquidity. The assemblies with arginine (RL) or lysine (KL) periphery exhibited positive charges (∼+35 mV) and complete condensation of pDNA into nanosized complexes (∼120 nm). In contrast, assemblies composed of histidine-rich lipids (HL) showed relatively low cationic electric potential (∼+10 mV) and poor DNA binding ability. As expected, the designed RL assemblies with guanidine-rich periphery enhanced the in vitro gene transfection up to 190-fold as compared with the golden standard PEI25k and Lipofectamine 2000, especially in the presence of serum. Meanwhile, interaction with cell and endo/lysosome membrane also revealed the superiority of RL complexes, that the guanidine-rich surface efficiently promoted transmembrane process in cellular internalization and endosomal disruption. More importantly, RL complexes also succeeded beyond others in vivo with significantly (∼7-fold) enhanced expression in HepG2 tumor xenografts in mice, as well as stronger green fluorescence protein imaging in isolated tumors and tumor frozen sections.
基于阳离子脂质的组装体为将基因有效浓缩成纳米颗粒提供了一个有前景的平台，并且组装体的外围性质对于与物理屏障的络合和相互作用至关重要。在此，我们报道了三种阳离子双头脂质，它们各自含有一个二油酰谷氨酸疏水尾部和一个由赖氨酸、精氨酸或组氨酸组成的双极性头部。已证明此类脂质在水溶液中能自组装形成具有明确纳米结构且外围富含氨基、胍基或咪唑的结构，显示出强大的缓冲能力和良好的流动性。具有精氨酸（RL）或赖氨酸（KL）外围的组装体呈现正电荷（约 +35 mV），并能将 pDNA 完全浓缩成纳米级复合物（约 120 nm）。相比之下，由富含组氨酸的脂质（HL）组成的组装体显示出相对较低的阳离子电势（约 +10 mV）和较差的 DNA 结合能力。正如预期的那样，与金标准 PEI25k 和 Lipofectamine 2000 相比，设计的具有富含胍基外围的 RL 组装体将体外基因转染效率提高了 190 倍，尤其是在有血清存在的情况下。同时，与细胞和内体/溶酶体膜的相互作用也揭示了 RL 复合物的优势，即富含胍基的表面在细胞内化和内体破坏过程中有效地促进了跨膜过程。更重要的是，RL 复合物在体内也比其他复合物更成功，在小鼠 HepG2 肿瘤异种移植中表达显著增强（约 7 倍），并且在分离的肿瘤和肿瘤冰冻切片中绿色荧光蛋白成像更强。
