Overcoming endosomal barrier by amphotericin B-loaded dual pH-responsive PDMA-b-PDPA micelleplexes for siRNA delivery.
Overcoming endosomal barrier by amphotericin B-loaded dual pH-responsive PDMA-b-PDPA micelleplexes for siRNA delivery.
复制标题
通过两性霉素B负载双pH响应PDMA-B-PDPA胶束胶束来克服内体屏障，用于siRNA递送。
DOI：
10.1021/nn203503h
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发表时间：
2011-11-22
期刊：
影响因子：
17.1
通讯作者：
Gao J
中科院分区：
文献类型：
--
作者：
Yu H;Zou Y;Wang Y;Huang X;Huang G;Sumer BD;Boothman DA;Gao J
内体屏障是非病毒纳米载体在细胞内有效传递siRNA的主要瓶颈。在这里，我们报道了一种新型的两性霉素B(AmB)，双pH响应型胶束复合体平台，用于siRNA的传递。以聚(2-(二甲氨基)乙基-甲基丙烯酸乙酯)(methacrylate)-block-poly(2-(diisopropylamino)ethyl-b-PDPA)两嵌段共聚物自组装成胶束。在pH 7.4时，Amb被负载到疏水的PDPA核心中，siRNA与带正电的PDMA壳层络合形成胶束。细胞摄取后，PDMA-b-PDPA/siRNA胶束在早期内吞体内解离，释放Amb。活细胞成像研究表明，释放的Amb显著增加了siRNA克服内体屏障的能力。转染研究表明，与不含AmB的对照相比，负载AmB的胶束复合体显著提高了荧光素酶(Luc)的敲除效率。这种增强的Luc基因敲除效率可被Bafilmycin A1消除，后者是一种空泡ATPase抑制剂，可抑制内吞细胞器的酸化。这些数据支持这一中心假设，即Amb的膜穿孔和一种“质子海绵”聚合物增加内体肿胀和膜张力提供了一种协同策略，以破坏内体以改善siRNA的细胞内递送。
The endosomal barrier is a major bottleneck for the effective intracellular delivery of siRNA by nonviral nanocarriers. Here, we report a novel amphotericin B (AmB)-loaded, dual pH-responsive micelleplex platform for siRNA delivery. Micelles were self-assembled from poly(2-(dimethylamino)ethyl methacrylate)-block-poly(2-(diisopropylamino)ethyl methacrylate) (PDMA-b-PDPA) diblock copolymers. At pH 7.4, AmB was loaded into the hydrophobic PDPA core, and siRNA was complexed with a positively charged PDMA shell to form the micelleplexes. After cellular uptake, the PDMA-b-PDPA/siRNA micelleplexes dissociated in early endosomes to release AmB. Live cell imaging studies demonstrated that released AmB significantly increased the ability of siRNA to overcome the endosomal barrier. Transfection studies showed that AmB-loaded micelleplexes resulted in significant increase in luciferase (Luc) knockdown efficiency over the AmB-free control. The enhanced Luc knockdown efficiency was abolished by bafilomycin A1, a vacuolar ATPase inhibitor that inhibits the acidification of the endocytic organelles. These data support the central hypothesis that membrane poration by AmB and increased endosomal swelling and membrane tension by a “proton sponge” polymer provided a synergistic strategy to disrupt endosomes for improved intracellular delivery of siRNA.