Scavenger receptor-mediated uptake of cell-penetrating peptide nanocomplexes with oligonucleotides
Scavenger receptor-mediated uptake of cell-penetrating peptide nanocomplexes with oligonucleotides
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DOI：
10.1096/fj.11-191536
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发表时间：
2012-03-01
期刊：
影响因子：
4.8
通讯作者：
Langel, Ulo
中科院分区：
文献类型：
--
作者：
Ezzat, Kariem;Helmfors, Henrik;Langel, Ulo
细胞穿透肽 (CPP) 是短的阳离子肽，通过与带负电的质膜相互作用来穿透细胞；然而，详细的吸收机制尚不清楚。与 CPP 的传统作用模式相反，我们在此表明 CPP PepFect14 (PF14) 与寡核苷酸形成带负电荷的纳米复合物，并且它们的摄取是由 A 类清道夫受体 (SCARA) 介导的。 SCARA 的特异性抑制配体，例如岩藻多糖、聚肌苷酸和硫酸葡聚糖，完全抑制 HeLa pLuc705 剪接校正细胞模型中 PF14-寡核苷酸纳米复合物的活性，而非特异性化学相关分子则不会。此外，该细胞系中表达的 SCARA 亚型（SCARA3 和 SCARA5）的 RNA 干扰 (RNAi) 敲低导致活性显着降低至 < 50%。与此一致，免疫染色显示纳米复合物与受体普遍共定位，并且电子显微镜图像显示在抑制性配体存在的情况下纳米复合物没有结合或内化。有趣的是，当高浓度使用时，裸寡核苷酸也与 SCARA 共定位。这些结果证明 SCARA3 和 SCARA5 参与 PF14-寡核苷酸纳米复合物的摄取，并首次表明一些基于 CPP 的系统通过清道夫受体发挥作用，这可能为理解和改善 CPP 转染提供新的可能性。-Ezzat, K.、Helmfors, H.、Tudoran, O.、Juks, C.、Lindberg, S.、Padari, K., El-Andaloussi，S.，Pooga，M.，Langel，U.清道夫受体介导的细胞穿透肽纳米复合物与寡核苷酸的摄取。 FASEB J. 26, 1172-1180 (2012)。 www.fasebj.org
Cell-penetrating peptides (CPPs) are short cationic peptides that penetrate cells by interacting with the negatively charged plasma membrane; however, the detailed uptake mechanism is not clear. In contrary to the conventional mode of action of CPPs, we show here that a CPP, PepFect14 (PF14), forms negatively charged nanocomplexes with oligonucleotides and their uptake is mediated by class-A scavenger receptors (SCARAs). Specific inhibitory ligands of SCARAs, such as fucoidin, polyinosinic acid, and dextran sulfate, totally inhibit the activity of PF14-oligonucleotide nanocomplexes in the HeLa pLuc705 splice-correction cell model, while nonspecific, chemically related molecules do not. Furthermore, RNA interference (RNAi) knockdown of SCARA subtypes (SCARA3 and SCARA5) that are expressed in this cell line led to a significant reduction of the activity to < 50%. In line with this, immunostaining shows prevalent colocalization of the nanocomplexes with the receptors, and electron microscopy images show no binding or internalization of the nanocomplexes in the presence of the inhibitory ligands. Interestingly, naked oligonucleotides also colocalize with SCARAs when used at high concentrations. These results demonstrate the involvement of SCARA3 and SCARA5 in the uptake of PF14-oligonucleotide nanocomplexes and suggest for the first time that some CPP-based systems function through scavenger receptors, which could yield novel possibilities to understand and improve the transfection by CPPs.-Ezzat, K., Helmfors, H., Tudoran, O., Juks, C., Lindberg, S., Padari, K., El-Andaloussi, S., Pooga, M., Langel, U. Scavenger receptor-mediated uptake of cell-penetrating peptide nanocomplexes with oligonucleotides. FASEB J. 26, 1172-1180 (2012). www.fasebj.org