Mbatha Londiwe Simphiwe, Maiyo Fiona, Daniels Aliscia, Singh Moganavelli
Nano-Gene and Drug Delivery Group, Discipline of Biochemistry, School of Life Sciences, University of KwaZulu-Natal, Private Bag X54001, Durban 4000, South Africa.
Pharmaceutics. 2021 Jun 17;13(6):900. doi: 10.3390/pharmaceutics13060900.
Messenger RNA (mRNA) is not an attractive candidate for gene therapy due to its instability and has therefore received little attention. Recent studies show the advantage of mRNA over DNA, especially in cancer immunotherapy and vaccine development. This study aimed to formulate folic-acid-(FA)-modified, poly-amidoamine-generation-5 (PAMAM G5D)-grafted gold nanoparticles (AuNPs) and to evaluate their cytotoxicity and transgene expression using the luciferase reporter gene (F-mRNA) in vitro. Nanocomplexes were spherical and of favorable size. Nanocomplexes at optimum nanoparticle:mRNA (/) binding ratios showed good protection of the bound mRNA against nucleases and were well tolerated in all cell lines. Transgene expression was significantly ( < 0.0001) higher with FA-targeted, dendrimer-grafted AuNPs (Au:G5D:FA) in FA receptors overexpressing MCF-7 and KB cells compared to the G5D and G5D:FA NPs, decreasing significantly ( < 0.01) in the presence of excess competing FA ligand, which confirmed nanocomplex uptake via receptor mediation. Overall, transgene expression of the Au:G5D and Au:G5D:FA nanocomplexes exceeded that of G5D and G5D:FA nanocomplexes, indicating the pivotal role played by the inclusion of the AuNP delivery system. The favorable properties imparted by the AuNPs potentiated an increased level of luciferase gene expression.
信使核糖核酸（mRNA）由于其不稳定性，并非基因治疗的理想候选者，因此很少受到关注。最近的研究显示了mRNA相对于DNA的优势，尤其是在癌症免疫治疗和疫苗开发方面。本研究旨在制备叶酸（FA）修饰的、接枝了第5代聚酰胺-胺（PAMAM G5D）的金纳米颗粒（AuNP），并在体外使用荧光素酶报告基因（F-mRNA）评估其细胞毒性和转基因表达。纳米复合物呈球形且尺寸适宜。纳米颗粒与mRNA的最佳结合比例（/）下的纳米复合物对结合的mRNA具有良好的核酸酶保护作用，并且在所有细胞系中耐受性良好。与G5D和G5D:FA纳米颗粒相比，在过表达FA受体的MCF-7和KB细胞中，FA靶向的、树枝状聚合物接枝的AuNP（Au:G5D:FA）的转基因表达显著更高（<0.0001），在存在过量竞争性FA配体时显著降低（<0.01），这证实了纳米复合物通过受体介导的摄取。总体而言，Au:G5D和Au:G5D:FA纳米复合物的转基因表达超过了G5D和G5D:FA纳米复合物，表明包含AuNP递送系统所起的关键作用。AuNP赋予的良好特性增强了荧光素酶基因表达水平。
