Huysmans Hanne, Zhong Zifu, De Temmerman Joyca, Mui Barbara L, Tam Ying K, Mc Cafferty Séan, Gitsels Arlieke, Vanrompay Daisy, Sanders Niek N
Laboratory of Gene Therapy, Department of Nutrition, Genetics and Ethology, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium.
Laboratory of Gene Therapy, Department of Nutrition, Genetics and Ethology, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium; Department of Pathology, Bacteriology and Poultry Diseases, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium.
Mol Ther Nucleic Acids. 2019 Sep 6;17:867-878. doi: 10.1016/j.omtn.2019.08.001. Epub 2019 Aug 7.
In this work, we studied the expression kinetics and innate immune response of a self-amplifying mRNA (sa-RNA) after electroporation and lipid-nanoparticle (LNP)-mediated delivery in the skin of mice. Intradermal electroporation of the sa-RNA resulted in a plateau-shaped expression, with the plateau between day 3 and day 10. The overall protein expression of sa-RNA was significantly higher than that obtained after electroporation of plasmid DNA (pDNA) or non-replication mRNAs. Moreover, using IFN-β reporter mice, we elucidated that intradermal electroporation of sa-RNA induced a short-lived moderate innate immune response, which did not affect the expression of the sa-RNA. A completely different expression profile and innate immune response were observed when LNPs were used. The expression peaked 24 h after intradermal injection of sa-RNA-LNPs and subsequently showed a sharp drop. This drop might be explained by a translational blockage caused by the strong innate immune response that we observed in IFN-β reporter mice shortly (4 h) after intradermal injection of sa-RNA-LNPs. A final interesting observation was the capacity of sa-RNA-LNPs to transfect the draining lymph nodes after intradermal injection.
在本研究中，我们研究了自扩增mRNA（sa-RNA）在小鼠皮肤中经电穿孔和脂质纳米颗粒（LNP）介导递送后的表达动力学和先天免疫反应。sa-RNA的皮内电穿孔导致表达呈平台状，平台期在第3天至第10天之间。sa-RNA的总体蛋白表达显著高于质粒DNA（pDNA）或非复制mRNA电穿孔后获得的表达。此外，使用IFN-β报告基因小鼠，我们阐明了sa-RNA的皮内电穿孔诱导了短暂的中度先天免疫反应，这并不影响sa-RNA的表达。当使用LNP时，观察到了完全不同的表达谱和先天免疫反应。在皮内注射sa-RNA-LNP后24小时表达达到峰值，随后急剧下降。这种下降可能是由于我们在皮内注射sa-RNA-LNP后不久（4小时）在IFN-β报告基因小鼠中观察到的强烈先天免疫反应导致的翻译阻断。最后一个有趣的观察结果是sa-RNA-LNP在皮内注射后转染引流淋巴结的能力。
