Cell-penetrating peptide-conjugated lipid nanoparticles for siRNA delivery
Cell-penetrating peptide-conjugated lipid nanoparticles for siRNA delivery
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DOI：
10.1016/j.bbrc.2014.01.107
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发表时间：
2014-02-21
影响因子：
3.1
通讯作者：
Oku, Naoto
中科院分区：
文献类型：
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作者：
Asai, Tomohiro;Tsuzuku, Takuma;Oku, Naoto
制备了细胞穿透肽修饰的脂质纳米粒（LNP），用于向细胞内递送小干扰RNA（siRNA）。以鱼精蛋白为原料，合成了CPP的脂质衍生物，并将其用于制备CPP修饰的LNP（CPP-LNP）。将siRNA包封到CPP-LNP中提高了siRNA在血清中的稳定性。在CPP-LNP中配制的荧光标记的siRNA以时间依赖性的方式有效地内化到B16 F10鼠黑素瘤细胞中，尽管在没有CPP的LNP中几乎没有内化到这些细胞中。在用CPP-LNP中的siRNA转染的细胞中，大部分siRNA分布在这些细胞的细胞质中，而不在溶酶体中定位。内吞途径的分析表明，CPP-LNP主要通过巨胞饮和硫酸乙酰肝素介导的内吞作用内化。在表达荧光素酶的B16 F10细胞和表达增强型绿色荧光蛋白的HT 1080人纤维肉瘤细胞中，封装siRNA的CPP-LNP有效地诱导RNA干扰介导的报告基因沉默。这些数据表明，用鱼精蛋白衍生的CPP修饰LNP可有效促进siRNA在细胞质中的内化，从而增强基因沉默。(C)2014 Elsevier Inc. All rights reserved.
Lipid nanoparticles (LNP) modified with cell-penetrating peptides (CPP) were prepared for the delivery of small interfering RNA (siRNA) into cells. Lipid derivatives of CPP derived from protamine were newly synthesized and used to prepare CPP-decorated LNP (CPP-LNP). Encapsulation of siRNA into CPP-LNP improved the stability of the siRNA in serum. Fluorescence-labeled siRNA formulated in CPP-LNP was efficiently internalized into B16F10 murine melanoma cells in a time-dependent manner, although that in LNP without CPP was hardly internalized into these cells. In cells transfected with siRNA in CPP-LNP, most of the siRNA was distributed in the cytoplasm of these cells and did not localize in the lysosomes. Analysis of the endocytotic pathway indicated that CPP-LNP were mainly internalized via macropinocytosis and heparan sulfate-mediated endocytosis. CPP-LNP encapsulating siRNA effectively induced RNA interference-mediated silencing of reporter genes in B16F10 cells expressing luciferase and in HT1080 human fibrosarcoma cells expressing enhanced green fluorescent protein. These data suggest that modification of LNP with the protamine-derived CPP was effective to facilitate internalization of siRNA in the cytoplasm and thereby to enhance gene silencing. (C) 2014 Elsevier Inc. All rights reserved.