Highly efficient siRNA transfection in macrophages using apoptotic body-mimic Ca-PS lipopolyplex.
Highly efficient siRNA transfection in macrophages using apoptotic body-mimic Ca-PS lipopolyplex.
复制标题
使用模拟凋亡体 Ca-PS 脂多聚复合物在巨噬细胞中高效转染 siRNA
DOI：
10.2147/ijn.s176991
复制
发表时间：
2018
影响因子：
8
通讯作者：
Hua Z
中科院分区：
文献类型：
--
作者：
Lai Y;Xu X;Zhu Z;Hua Z
RNA干扰的发现和发展为生物化学和生物医学领域做出了巨大贡献。然而，由于转染效率极低，将sirna传递到某些类型的细胞（如免疫细胞）的脂质体转染方案是不可行的。虽然病毒传递和电穿孔是转染免疫细胞的两种广泛采用的方法，但它们存在一些缺点，如制备的复杂性、生物安全性问题和高细胞毒性。我们相信可以对脂质体配方和方案进行修改，以实现脂质体负载sirna的高效基因敲低。本研究的目的是利用凋亡模拟Ca-PS脂复合物在难以转染的巨噬细胞中实现高效的siRNA基因敲低，从而降低细胞毒性和提高细胞摄取效率。结果我们设计了一种含有磷脂酰丝氨酸的阴离子脂质体配方来模拟凋亡小体Ca-PS脂复合物。Ca-PS脂复合物被证明能够在降低细胞毒性的情况下，在多种细胞系中传递和影响有效的基因敲除。在Ana-1和骨髓源性巨噬细胞两种类型的巨噬细胞中，Ca-PS脂复合物的敲除效率比Lipo2000提高了157%。进一步的研究表明，Ca-PS促进巨噬细胞的细胞摄取，溶酶体逃逸和sirna定位到核周区域。最后，Ca-PS脂复合物转染不会诱导巨噬细胞自发极化。结论凋亡模拟体Ca-PS脂复合物是一种稳定的、无细胞毒性的sirna脂质体递送系统，其对巨噬细胞的效力大大提高，细胞毒性降低。推测Ca-PS脂复合物可应用于其他免疫细胞，如T细胞和DC细胞，但需要进一步的研究来探索其前景。
Background The discovery and development of RNA interference has made a tremendous contribution to the biochemical and biomedical field. However, liposomal transfection protocols to deliver siRNAs to certain types of cells, eg, immune cells, are not viable due to exceedingly low transfection efficiency. While viral delivery and electroporation are two widely adopted approaches to transfect immune cells, they are associated with certain drawbacks such as complexity of preparation, biosafety issues, and high cytotoxicity. We believe amendments can be made to liposomal formulas and protocols to achieve a highly efficient knockdown of genes by liposome-loaded siRNAs. Aim The aim of this study was to use the apoptotic-mimic Ca-PS lipopolyplex to achieve highly efficient siRNA knockdown of genes in the hard-to-transfect macrophages with reduced cytotoxicity and more efficient cellular uptake. Results We devised an anionic liposomal formula containing phosphatidylserine to mimic the apoptotic body, the Ca-PS lipopolyplex. Ca-PS lipopolyplex was proven to be capable of delivering and effecting efficient gene knockdown in multiple cell lines at lowered cytotoxicity. Among the two types of macrophages, namely Ana-1 and bone-marrow derived macrophages, Ca-PS lipopolyplex showed an improvement in knockdown efficiency, as high as 157%, over Lipo2000. Further investigations revealed that Ca-PS promotes increased cellular uptake, lysosomal escape and localization of siRNAs to the perinuclear regions in macrophages. Lastly, transfection by Ca-PS lipopolyplex did not induce spontaneous polarization of macrophages. Conclusion The apoptotic body-mimic Ca-PS lipopolyplex is a stable, non-cytotoxic liposomal delivery system for siRNAs featuring vastly improved potency for macrophages and lowered cytotoxicity. It is speculated that Ca-PS lipopolyplex can be applied to other immune cells such as T cells and DC cells, but further research efforts are required to explore its promising potentials.