University of Coimbra, CNC-Center for Neuroscience and Cell Biology, Department of Life Sciences, Calçada Martim de Freitas, 3000-456, Coimbra, Portugal.
CNC- Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504, Coimbra, Portugal.
Pharm Res. 2020 Sep 4;37(10):188. doi: 10.1007/s11095-020-02904-x.
This study aimed to endow the cell-penetrating peptide (CPP) S4-PV with adequate features towards a safe and effective application in cancer gene therapy.
Peptide/siRNA complexes were prepared with two new derivatives of the CPP S4-PV, which combine a lauroyl group attached to the N- or C-terminus with a histidine-enrichment in the N-terminus of the S4-PV peptide, being named C12-H-S4-PV and H-S4-PV-C12, respectively. Physicochemical characterization of siRNA complexes was performed and their cytotoxicity and efficiency to mediate siRNA delivery and gene silencing in cancer cells were assessed in the absence and presence of serum.
Peptide/siRNA complexes prepared with the C12-H-S4-PV derivative showed a nanoscale (ca. 100 nm) particle size, as revealed by TEM, and efficiently mediated gene silencing (37%) in human U87 glioblastoma cells in the presence of 30% serum. In addition, the new C12-H-S4-PV-based siRNA delivery system efficiently downregulated stearoyl-CoA desaturase-1, a key-enzyme of lipid metabolism overexpressed in cancer, which resulted in a significant decrease in the viability of U87 cells. Importantly, these complexes were able to spare healthy human astrocytes.
These encouraging results pave the way for a potential application of the C12-H-S4-PV peptide as a promising tool in cancer gene therapy.
本研究旨在赋予穿膜肽（CPP）S4-PV 足够的特性，使其在癌症基因治疗中安全有效应用。
用 S4-PV 肽的两个新衍生物——在 N 端带有豆蔻酰基团和 N 端富含组氨酸的 C12-H-S4-PV 和 H-S4-PV-C12，制备肽/siRNA 复合物。对 siRNA 复合物进行理化性质表征，并评估其在无血清和有血清存在的情况下对癌细胞中 siRNA 递释和基因沉默的细胞毒性和效率。
通过 TEM 揭示，用 C12-H-S4-PV 衍生物制备的肽/siRNA 复合物具有纳米级（约 100nm）的粒径，并在 30%血清存在的情况下，有效介导人 U87 神经胶质瘤细胞中的基因沉默（37%）。此外，新型 C12-H-S4-PV 基 siRNA 递释系统可有效下调脂代谢中过表达的关键酶硬脂酰辅酶 A 去饱和酶-1，从而显著降低 U87 细胞的活力。重要的是，这些复合物能够保护健康的人星形胶质细胞。
这些令人鼓舞的结果为 C12-H-S4-PV 肽作为癌症基因治疗中一种有前途的工具的潜在应用铺平了道路。
