Influence of cationic lipid composition on gene silencing properties of lipid nanoparticle formulations of siRNA in antigen-presenting cells.
Influence of cationic lipid composition on gene silencing properties of lipid nanoparticle formulations of siRNA in antigen-presenting cells.
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DOI：
10.1038/mt.2011.190
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发表时间：
2011-12
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脂质纳米颗粒（LNPs）是目前使用小干扰RNA沉默肝细胞靶基因的最有效的体内递送系统。抗原呈递细胞（APCs）也是LNP siRNA的潜在靶标。我们检测了原代骨髓巨噬细胞（bmMΦ）和树突细胞中siRNA的摄取、细胞内运输和基因沉默效力，这些siRNA是在含有四种不同的可电离阳离子脂质的LNPs中形成的，即DLinDAP、DLinDMA、DLinK-DMA和DLinKC2-DMA。含有DLinKC2-DMA的LNPs是最有效的配方，因为它们能够抑制GAPDH靶蛋白的产生。此外，含有DLinKC2-DMA的LNPs是最有效的细胞内递送剂，使用荧光标记的siRNA对内体和细胞质siRNA定位的共聚焦研究表明。与DLinDMA相比，dlinkdma和DLinKC2-DMA配方表现出更好的基因沉默能力，但毒性较小。体内实验结果表明，含有DLinKC2-DMA的LNP siRNA系统在系统给药后可以有效地沉默脾脏和腹腔APCs中的GAPDH。APCs中的基因沉默是RNAi介导的，使用较大的LNPs可显著减少肝细胞沉默，而APCs中也保持了类似的效果。这些结果讨论了LNP siRNA制剂治疗免疫介导疾病的潜力。
Lipid nanoparticles (LNPs) are currently the most effective in vivo delivery systems for silencing target genes in hepatocytes employing small interfering RNA. Antigen-presenting cells (APCs) are also potential targets for LNP siRNA. We examined the uptake, intracellular trafficking, and gene silencing potency in primary bone marrow macrophages (bmMΦ) and dendritic cells of siRNA formulated in LNPs containing four different ionizable cationic lipids namely DLinDAP, DLinDMA, DLinK-DMA, and DLinKC2-DMA. LNPs containing DLinKC2-DMA were the most potent formulations as determined by their ability to inhibit the production of GAPDH target protein. Also, LNPs containing DLinKC2-DMA were the most potent intracellular delivery agents as indicated by confocal studies of endosomal versus cytoplamic siRNA location using fluorescently labeled siRNA. DLinK-DMA and DLinKC2-DMA formulations exhibited improved gene silencing potencies relative to DLinDMA but were less toxic. In vivo results showed that LNP siRNA systems containing DLinKC2-DMA are effective agents for silencing GAPDH in APCs in the spleen and peritoneal cavity following systemic administration. Gene silencing in APCs was RNAi mediated and the use of larger LNPs resulted in substantially reduced hepatocyte silencing, while similar efficacy was maintained in APCs. These results are discussed with regard to the potential of LNP siRNA formulations to treat immunologically mediated diseases.