Dept. of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 26, 6525 GA, Nijmegen, The Netherlands; Dept. of Obstetrics and Gynaecology, Radboud University Medical Center, Geert Grooteplein 10, 6525 GA, Nijmegen, The Netherlands.
Dept. of Cell Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein 26, 6525 GA, Nijmegen, The Netherlands.
Biochim Biophys Acta Biomembr. 2018 Jun;1860(6):1342-1349. doi: 10.1016/j.bbamem.2018.03.007. Epub 2018 Mar 14.
Drug delivery into tumors and metastases is a major challenge in the eradication of cancers such as epithelial ovarian carcinoma. Cationic cell-penetrating peptides (CPPs) are a promising group of delivery vehicles to mediate cellular entry of molecules that otherwise poorly enter cells. However, little is known about their penetration behavior in tissues. Here, we investigated penetration of cationic CPPs in 3D ovarian cancer spheroids and patient-derived 3D tumor explants. Penetration kinetics and distribution after long-term incubation were imaged by confocal microscopy. In addition, spheroids and tumor explants were dissociated and cell-associated fluorescence determined by flow cytometry. CPPs with high uptake activity showed enhanced sequestration in the periphery of the spheroid, whereas less active CPPs were able to penetrate deeper into the tissue. CPPs consisting of d-amino acids were advantageous over l-amino acid CPPs as they showed less but long lasting cellular uptake activity, which benefitted penetration and retention over time. In primary tumor cultures, in contrast to nonaarginine, the amphipathic CPP penetratin was strongly sequestered by cell debris and matrix components pointing towards arginine-rich CPPs as a preferred choice. Overall, the data show that testing in 3D models leads to a different choice of the preferred peptide in comparison to a standard 2D cell culture.
将药物递送到肿瘤和转移灶是根除上皮性卵巢癌等癌症的主要挑战。阳离子细胞穿透肽（CPPs）是一类很有前途的递药载体，可介导其他不易进入细胞的分子进入细胞。然而，人们对它们在组织中的穿透行为知之甚少。在这里，我们研究了阳离子 CPP 在 3D 卵巢癌球体和患者来源的 3D 肿瘤外植体中的穿透行为。通过共聚焦显微镜对长期孵育后的穿透动力学和分布进行了成像。此外，还通过流式细胞术检测了球体和肿瘤外植体的解离和细胞相关荧光。摄取活性高的 CPP 表现出在球体边缘的强烈隔离，而活性较低的 CPP 则能够更深地穿透组织。由 d-氨基酸组成的 CPP 比 l-氨基酸 CPP 具有优势，因为它们的细胞摄取活性较低但持续时间较长，这有利于随着时间的推移穿透和保留。在原发性肿瘤培养物中，与非精氨酸相反，两亲性 CPP penetratin 被细胞碎片和基质成分强烈隔离，这表明富含精氨酸的 CPP 是首选。总的来说，数据表明与标准 2D 细胞培养相比，在 3D 模型中进行测试会导致对首选肽的不同选择。
