One-step encapsulation of siRNA between lipid-layers of multi-layer polycation liposomes by lipoplex freeze-thawing
One-step encapsulation of siRNA between lipid-layers of multi-layer polycation liposomes by lipoplex freeze-thawing
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DOI：
10.1016/j.jconrel.2016.01.032
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发表时间：
2016-04-28
影响因子：
10.8
通讯作者：
Oku, Naoto
中科院分区：
文献类型：
--
作者：
Koide, Hiroyuki;Okamoto, Ayaka;Oku, Naoto
小干扰RNA(SiRNA)有可能成为治疗难治性疾病的候选药物。然而，由于无载体的siRNA的转染率较低，且在体内容易降解，因此需要合适的载体来传递siRNA。在这里，我们报道了一种简单、只需一步就能通过冻融将siRNA包裹到脂类纳米载体中的方法：通过冻融由聚阳离子脂质体(PCL)和siRNA组成的脂合物，将siRNA包裹在多层脂质体的脂层之间。通过重复冻融脂复合体5个循环，提高了对PCL的siRNA持有量。常规脂复合体中的siRNA在90%胎牛血清中孵育72 h后被降解，但冷冻和解冻后的脂复合体中的siRNA不被降解。有趣的是，我们发现，单层PCL与siRNA复合体冻融后，脂复合体形成了“堆积的多层”结构，这表明siRNA存在于脂层之间，起到了结合的作用。与常规脂复合体相比，冷冻和解冻后的脂复合体显示出显著更高的基因敲除效率。此外，与聚乙二醇化的传统脂质体相比，聚乙二醇化的冻融脂合物在体内向肿瘤传递了更多的siRNA。这些结果为通过冻融将siRNA“一步”包裹到脂质体中提供了一种有吸引力的策略。(C)2016爱思唯尔B.V.保留所有权利。
Small interfering RNA (siRNA) has the potential to be a candidate as a cure for intractable diseases. However, an appropriate vector is required for siRNA delivery because of the low transfection efficiency of siRNA without a vector and its easy degradation in vivo. Here, we report a simple, only one step, and efficient method for siRNA encapsulation into a lipidic nanocarrier by freeze-thawing: siRNA was entrapped between the lipid layers of multi-layer liposomes by freeze-thawing of lipoplexes composed of polycation liposomes (PCLs) and siRNA. siRNA-holding capacity to the PCL was increased by repeating freeze-thaw of the lipoplex up to 5 cycles. Although siRNA in the conventional lipoplex was degraded after incubation in 90% fetal bovine serum for 72 h, siRNA in the frozen and thawed lipoplex was not degraded. Interestingly, we found that the lipoplex formed a "packed multi-layer" structure after the freeze-thawing of "single-layer" PCL and siRNA complex, suggesting that siRNA exists between the lipid layers working as a binder. The frozen and thawed lipoplex showed significantly higher knockdown efficacy compared with the conventional lipoplex. In addition, PEGylated freeze-thawed lipoplexes delivered a higher amount of siRNA to a tumor in vivo compared with the PEGylated conventional ones. These results provide an attractive strategy for "one-step" encapsulation of siRNA into liposomes by freeze-thawing. (C) 2016 Elsevier B.V. All rights reserved.