Zeng Ye, Shen Mengjie, Pattipeiluhu Roy, Zhou Xuequan, Zhang Yun, Bakkum Thomas, Sharp Thomas H, Boyle Aimee L, Kros Alexander
Department of Supramolecular & Biomaterials Chemistry, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Nanoscale. 2023 Sep 29;15(37):15206-15218. doi: 10.1039/d3nr02175k.
Gene delivery has great potential in modulating protein expression in specific cells to treat diseases. Such therapeutic gene delivery demands sufficient cellular internalization and endosomal escape. Of various nonviral nucleic acid delivery systems, lipid nanoparticles (LNPs) are the most advanced, but still, are very inefficient as the majority are unable to escape from endosomes/lysosomes. Here, we develop a highly efficient gene delivery system using fusogenic coiled-coil peptides. We modified LNPs, carrying EGFP-mRNA, and cells with complementary coiled-coil lipopeptides. Coiled-coil formation between these lipopeptides induced fast nucleic acid uptake and enhanced GFP expression. The cellular uptake of coiled-coil modified LNPs is likely driven by membrane fusion thereby omitting typical endocytosis pathways. This direct cytosolic delivery circumvents the problems commonly observed with the limited endosomal escape of mRNA. Therefore fusogenic coiled-coil peptide modification of existing LNP formulations to enhance nucleic acid delivery efficiency could be beneficial for several gene therapy applications.
基因递送在调节特定细胞中的蛋白质表达以治疗疾病方面具有巨大潜力。这种治疗性基因递送需要足够的细胞内化和内体逃逸。在各种非病毒核酸递送系统中，脂质纳米颗粒（LNP）是最先进的，但仍然效率非常低，因为大多数无法从内体/溶酶体中逃逸。在这里，我们使用融合卷曲螺旋肽开发了一种高效的基因递送系统。我们用携带增强型绿色荧光蛋白（EGFP）-mRNA的LNP和具有互补卷曲螺旋脂肽的细胞进行修饰。这些脂肽之间的卷曲螺旋形成诱导了快速的核酸摄取并增强了绿色荧光蛋白的表达。卷曲螺旋修饰的LNP的细胞摄取可能是由膜融合驱动的，从而省略了典型的内吞途径。这种直接的胞质递送避免了通常观察到的mRNA内体逃逸受限的问题。因此，对现有LNP制剂进行融合卷曲螺旋肽修饰以提高核酸递送效率可能对几种基因治疗应用有益。
