Batta Gyula, Kárpáti Levente, Henrique Gabriela Fulaneto, Tóth Gabriella, Tarapcsák Szabolcs, Kovacs Tamas, Zakany Florina, Mándity István M, Nagy Peter
Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.
Department of Genetics and Applied Microbiology, Faculty of Science and Technology, University of Debrecen, Debrecen, Hungary.
Br J Pharmacol. 2021 Sep;178(18):3667-3681. doi: 10.1111/bph.15509. Epub 2021 May 27.
Cell penetrating peptides are promising tools for delivery of cargo into cells, but factors limiting or facilitating their cellular uptake are largely unknown. We set out to study the effect of the biophysical properties of the cell membrane on the uptake of penetratin, a cell penetrating peptide.
Using labelling with pH-insensitive and pH-sensitive dyes, the kinetics of cellular uptake and endo-lysosomal escape of penetratin were studied by flow cytometry.
We report that escape of penetratin from acidic endo-lysosomal compartments is retarded compared with its total cellular uptake. The membrane dipole potential, known to alter transmembrane transport of charged molecules, is shown to be negatively correlated with the concentration of penetratin in the cytoplasmic compartment. Treatment of cells with therapeutically relevant concentrations of atorvastatin, an inhibitor of HMG-CoA reductase and cholesterol synthesis, significantly increased endosomal escape of penetratin in two different cell types. This effect of atorvastatin correlated with its ability to decrease the membrane dipole potential.
These results highlight the importance of the dipole potential in regulating cellular uptake of cell penetrating peptides and suggest a clinically relevant way of boosting this process.
细胞穿透肽是将货物递送至细胞内的有前景的工具，但限制或促进其细胞摄取的因素很大程度上尚不清楚。我们着手研究细胞膜的生物物理特性对穿膜肽（一种细胞穿透肽）摄取的影响。
使用对pH不敏感和对pH敏感的染料进行标记，通过流式细胞术研究穿膜肽的细胞摄取动力学和内吞溶酶体逃逸情况。
我们报告称，与穿膜肽的总细胞摄取相比，其从酸性内吞溶酶体区室的逃逸受到阻碍。已知会改变带电分子跨膜转运的膜偶极电势，与细胞质区室中穿膜肽的浓度呈负相关。用治疗相关浓度的阿托伐他汀（一种HMG-CoA还原酶和胆固醇合成抑制剂）处理细胞，可显著增加两种不同细胞类型中穿膜肽的内体逃逸。阿托伐他汀的这种作用与其降低膜偶极电势的能力相关。
这些结果突出了偶极电势在调节细胞穿透肽细胞摄取中的重要性，并提示了一种促进该过程的临床相关方法。
