Mével Mathieu, Haudebourg Thomas, Colombani Thibault, Peuziat Pauline, Dallet Laurence, Chatin Benoît, Lambert Olivier, Berchel Mathieu, Montier Tristan, Jaffrès Paul-Alain, Lehn Pierre, Pitard Bruno
Unité INSERM UMR 1087, CNRS UMR 6291, Nantes, France.
Université de Nantes, l'institut du thorax, Nantes, France.
J Gene Med. 2016 Jan-Mar;18(1-3):3-15. doi: 10.1002/jgm.2869.
To optimize synthetic gene delivery systems, there is a need to develop more efficient lipid formulations. Most cationic lipid formulations contain 'helper' neutral lipids because of their ability to increase DNA delivery, in particular by improving endosomal escape of DNA molecules via the pH-buffering effect of protonatable groups and/or fusion with the lipid bilayer of endosomes.
We evaluated the influence of the linker structure between the two oleyl chains in the helper lipid on transfection efficiency in cell lines, as well as in primary cells (hepatocytes/cardiomyocytes). We reported the synthesis of two new pH-buffering imidazole helper lipids characterized by a polar headgroup containing one (compound 6) or two (compound 5) imidazole groups and two oleyl chains linked by an amide group. We studied their association with the aminoglycoside lipidic derivative dioleylsuccinylparomomycin (DOSP), which contains two oleyl chains linked to the aminoglycoside polar headgroup via an amide function. We compared the morphology and transfection properties of such binary liposomes of DOSP/5 and DOSP/6 with those of liposomes combining DOSP with another imidazole-based dioleyl helper lipid (MM27) in which a phosphoramido group acts as a linker between the two oleyl chains and imidazole function.
The phosphoramido linker in the helper lipid induces a major difference in terms of morphology and resistance to decomplexation at physical pH for DOSP/helper lipid complexes.
This hybrid dioleyl linker composition of DOSP/MM27 led to higher transfection efficiency in cell lines and in primary cells compared to complexes with homogeneous dioleyl linker.
为了优化合成基因递送系统，需要开发更有效的脂质制剂。大多数阳离子脂质制剂都含有“辅助”中性脂质，因为它们能够提高DNA递送效率，特别是通过可质子化基团的pH缓冲作用和/或与内体脂质双层融合来改善DNA分子的内体逃逸。
我们评估了辅助脂质中两条油酰链之间的连接结构对细胞系以及原代细胞（肝细胞/心肌细胞）转染效率的影响。我们报道了两种新型pH缓冲咪唑辅助脂质的合成，其特征在于极性头基团含有一个（化合物6）或两个（化合物5）咪唑基团以及通过酰胺基团连接的两条油酰链。我们研究了它们与氨基糖苷脂质衍生物二油酰琥珀酰巴龙霉素（DOSP）的缔合，DOSP包含通过酰胺官能团连接到氨基糖苷极性头基团的两条油酰链。我们将DOSP/5和DOSP/6的这种二元脂质体的形态和转染特性与将DOSP与另一种基于咪唑的二油酰辅助脂质（MM27）组合的脂质体进行了比较，其中磷酰胺基团充当两条油酰链和咪唑官能团之间的连接基团。
辅助脂质中的磷酰胺连接基团在DOSP/辅助脂质复合物的形态和对生理pH下解聚的抗性方面引起了重大差异。
与具有均匀二油酰连接基团的复合物相比，DOSP/MM27的这种混合二油酰连接基团组成在细胞系和原代细胞中导致更高的转染效率。
