School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, St. Lucia, QLD 4072, Australia. Department of Pharmaceutical Sciences and Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, 140 The Fenway, Boston, MA 02115, United States of America.
Nanotechnology. 2019 Oct 11;30(41):415604. doi: 10.1088/1361-6528/ab313d. Epub 2019 Jul 11.
The development of polymer-based nanoparticulate delivery systems for siRNA is important for the clinical success of gene therapy. However, there are some major drawbacks that need to be overcome. Short interfering RNA (siRNA) has been investigated as a potential therapeutic drug to silence disease-associated genes, but its usage is limited due to the lack of effective and safe nanocarriers. In this study, DOPE-PEI, a nanoparticle consisting of the fusogenic lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) conjugated with low-molecular-weight, 600 Da, branched polyethylenimine (PEI) was produced and optimized for siRNA delivery. This delivery system was modified with other components such as 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)2000] (DOPE-PEG2K), DOPE-PEG3.4K-bombesin and 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine/1,2-dioleoyl-3-trimethylammonium-propane (DOPE/DOTAP) and tested on PC-3 cells. The conjugation of DOPE to PEI polymer (DOPE-PEI) improved the efficiency of PEI to deliver siRNA into the cytosol and knockdown genes, but demonstrated high toxicity. The addition of DOPE-PEG2K reduced cellular toxicity by masking the surface positive charge of the DOPE-PEI/siRNA complex, with the incorporation of a gastrin-releasing peptide receptor (GRPR) targeting peptide and DOPE/DOTAP components improving the cellular uptake of siRNA into targeted cells and the siRNA knockdown efficiency.
聚合物基纳米颗粒递药系统的发展对于基因治疗的临床成功非常重要。然而，仍有一些主要的缺点需要克服。短干扰 RNA（siRNA）已被研究作为一种沉默疾病相关基因的潜在治疗药物，但由于缺乏有效和安全的纳米载体，其应用受到限制。在这项研究中，制备了一种由融合脂质 1,2-二油酰基-sn-甘油-3-磷酸乙醇胺（DOPE）与低分子量、600Da、支化聚乙烯亚胺（PEI）偶联而成的纳米颗粒 DOPE-PEI，并对其进行了优化以用于 siRNA 的递药。该递药系统经过其他成分的修饰，如 1,2-二油酰基-sn-甘油-3-磷酸乙醇胺-N-[甲氧基（聚乙二醇）2000]（DOPE-PEG2K）、DOPE-PEG3.4K-蛙皮素和 1,2-二油酰基-sn-甘油-3-磷酸乙醇胺/1,2-二油酰基-3-三甲基铵丙烷（DOPE/DOTAP），并在 PC-3 细胞上进行了测试。DOPE 与 PEI 聚合物（DOPE-PEI）的偶联提高了 PEI 将 siRNA 递送至细胞质并敲低基因的效率，但表现出高毒性。DOPE-PEG2K 的添加通过掩盖 DOPE-PEI/siRNA 复合物表面的正电荷来降低细胞毒性，而胃泌素释放肽受体（GRPR）靶向肽和 DOPE/DOTAP 成分的加入则提高了靶向细胞中 siRNA 的细胞摄取和 siRNA 敲低效率。
