Precise Engineering of siRNA Delivery Vehicles to Tumors Using Polyion Complexes and Gold Nanoparticles
Precise Engineering of siRNA Delivery Vehicles to Tumors Using Polyion Complexes and Gold Nanoparticles
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DOI：
10.1021/nn502125h
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发表时间：
2014-09-01
期刊：
影响因子：
17.1
通讯作者：
Kataoka, Kazunori
中科院分区：
文献类型：
--
作者：
Kim, Hyun Jin;Takemoto, Hiroyasu;Kataoka, Kazunori
为了将小干扰RNA（siRNA）系统性递送至实体瘤，利用载有20 kDa siRNA的单聚体聚离子复合物（uPIC）和20纳米金纳米粒子（AuNP）构建了一种尺寸可调控且可逆稳定的纳米结构。通过电荷匹配的聚离子络合作用选择性地制备uPIC，即带有约40个正电荷（且在ω端有巯基）的聚乙二醇 - b - 聚（L - 赖氨酸）（PEG - PLL）共聚物与带有40个负电荷的单个siRNA进行络合。PEG - PLL在ω端的巯基进一步使得uPIC能够通过配位键成功结合到单个AuNP上，生成尺寸为38纳米且尺寸分布窄的纳米结构（uPIC - AuNP）。相反，在没有siRNA的情况下，将巯基化的PEG - Pas和AuNPs混合会产生大的聚集体，这表明预先形成的uPIC在纳米结构形成中起着至关重要的作用。与单独的uPIC相比，智能的uPIC - AuNPs在含血清的培养基中稳定，并且对肝素诱导的反聚阴离子交换更具抵抗力。另一方面，用细胞内浓度的谷胱甘肽处理uPIC - AuNPs会显著降低其稳定性并触发siRNA的释放，这表明这些纳米结构相对于巯基交换和带负电的AuNP表面具有可逆稳定性。uPIC - AuNPs有效地将siRNA递送至培养的癌细胞中，促进了显著的序列特异性基因沉默且无细胞毒性。与对照组（即裸AuNPs和uPIC）相比，系统性给药的uPIC - AuNPs显示出明显更长的血液循环时间，这表明uPIC结合到AuNP上对于延长血液循环时间至关重要。最后，uPIC - AuNPs在皮下接种的表达荧光素酶的宫颈癌（HeLa - Luc）模型中高效聚集，并在肿瘤组织中实现了显著的荧光素酶基因沉默。这些结果表明uPIC - AuNP纳米结构在将siRNA系统性递送至实体瘤方面具有巨大潜力。
For systemic delivery of siRNA to solid tumors, a size-regulated and reversibly stabilized nanoarchitecture was constructed by using a 20 kDa siRNA-loaded unimer polyion complex (uPIC) and 20 nm gold nanoparticle (AuNP). The uPIC was selectively prepared by charge-matched polyionic complexation of a poly(ethylene glycol)-b-poly(L-lysine) (PEG-PLL) copolymer bearing similar to 40 positive charges (and thiol group at the omega-end) with a single siRNA bearing 40 negative charges. The thiol group at the omega-end of PEG-PLL further enabled successful conjugation of the uPICs onto the single AuNP through coordinate bonding, generating a nanoarchitecture (uPIC-AuNP) with a size of 38 nm and a narrow size distribution. In contrast, mixing thiolated PEG-Pas and AuNPs produced a large aggregate in the absence of siRNA, suggesting the essential role of the preformed uPIC in the formation of nanoarchitecture. The smart uPIC-AuNPs were stable in serum-containing media and more resistant against heparin-induced counter polyanion exchange, compared to uPICs alone. On the other hand, the treatment of uPIC-AuNPs with an intracellular concentration of glutathione substantially compromised their stability and triggered the release of siRNA, demonstrating the reversible stability of these nanoarchitectures relative to thiol exchange and negatively charged AuNP surface. The uPIC-AuNPs efficiently delivered siRNA into cultured cancer cells, facilitating significant sequence-specific gene silencing without cytotoxicity. Systemically administered uPIC-AuNPs showed appreciably longer blood circulation time compared to controls, i.e., bare AuNPs and uPICs, indicating that the conjugation of uPICs onto AuNP was crucial for enhancing blood circulation time. Finally, the uPIC-AuNPs efficiently accumulated in a subcutaneously inoculated luciferase-expressing cervical cancer (HeLa-Luc) model and achieved significant luciferase gene silencing in the tumor tissue. These results demonstrate the strong potential of uPIC-AuNP nanoarchitectures for systemic siRNA delivery to solid tumors.