Begum Anjuman Ara, Wan Yu, Toth Istvan, Moyle Peter M
School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia 4072, Australia; School of Pharmacy, The University of Queensland, Woolloongabba 4102, Australia.
School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia 4072, Australia; School of Pharmacy, The University of Queensland, Woolloongabba 4102, Australia; Institute of Molecular Bioscience, The University of Queensland, St Lucia 4072, Australia.
Bioorg Med Chem. 2018 Jan 15;26(2):516-526. doi: 10.1016/j.bmc.2017.12.013. Epub 2017 Dec 12.
The development of non-viral gene delivery systems, with the capacity to overcome most of the biological barriers facing gene delivery, is challenging. We have developed peptide-based, multicomponent, non-viral delivery systems, incorporating: a bombesin peptide ligand (BBN(6-14)), to selectively target the gastrin releasing peptide receptor (GRPR); oligoarginine peptides (hexa- (R) and nona-arginine (R)), for plasmid DNA (pDNA) condensation; and GALA, to facilitate endosome escape. The uptake and endosome escape efficiency of bombesin/oligoarginine and bombesin/oligoarginine/GALA fusion peptides for oligonucleotide delivery was evaluated in terms of their complex size, cellular uptake, endosome escape, and cellular toxicity. Complex size and cell uptake studies demonstrated that the nona-arginine/bombesin delivery system was more efficient at condensing and delivering pDNA into PC-3 prostate cancer cells compared to the hexa-arginine/bombesin delivery system. Further, competition with free bombesin peptide, and comparative uptake studies in Caco-2 cells, which express GRPR at a lower level, suggested that GRPR contributes to the targeted uptake of this system. The addition of GALA into the nona-arginine/bombesin-based system further increased the pDNA cellular uptake at all tested N/P ratios; facilitated endosomal pDNA release; and had limited effects on cell viability. In conclusion, the delivery system combining BBN(6-14) with nona-arginine and GALA had optimal characteristics for the delivery of pDNA into the GRPR overexpressing cell line PC-3.
开发能够克服基因递送所面临的大多数生物屏障的非病毒基因递送系统具有挑战性。我们已经开发了基于肽的多组分非病毒递送系统，该系统包含：一种蛙皮素肽配体（BBN(6 - 14)），用于选择性靶向胃泌素释放肽受体（GRPR）；寡聚精氨酸肽（六聚精氨酸（R）和九聚精氨酸（R）），用于质粒DNA（pDNA）凝聚；以及GALA，用于促进内体逃逸。通过其复合物大小、细胞摄取、内体逃逸和细胞毒性评估了蛙皮素/寡聚精氨酸和蛙皮素/寡聚精氨酸/GALA融合肽用于寡核苷酸递送的摄取和内体逃逸效率。复合物大小和细胞摄取研究表明，与六聚精氨酸/蛙皮素递送系统相比，九聚精氨酸/蛙皮素递送系统在将pDNA凝聚并递送至PC - 3前列腺癌细胞方面更有效。此外，与游离蛙皮素肽的竞争以及在低水平表达GRPR的Caco - 2细胞中的比较摄取研究表明，GRPR有助于该系统的靶向摄取。在基于九聚精氨酸/蛙皮素的系统中添加GALA进一步提高了在所有测试的N/P比下pDNA的细胞摄取；促进了内体pDNA释放；并且对细胞活力的影响有限。总之，将BBN(6 - 14)与九聚精氨酸和GALA结合的递送系统在将pDNA递送至GRPR过表达细胞系PC - 3方面具有最佳特性。
