Dalal Chumki, Jana Nikhil R
Centre for Advanced Materials, Indian Association for the Cultivation of Science , Kolkata 700032, India.
J Phys Chem B. 2017 Apr 13;121(14):2942-2951. doi: 10.1021/acs.jpcb.6b12182. Epub 2017 Mar 31.
Although trans-activating transcription (TAT) peptide-functionalized nanoparticle/polymer/liposome is widely used for cellular transfection applications, the multivalency (number of attached peptide per particle) effect on cell uptake mechanism and subcellular targeting performance is largely unexplored. Here we show that multivalency of nanoparticle controls the cellular interaction, cellular entry/exit mechanism, and subcellular targeting performance. We have synthesized TAT-peptide functionalized quantum dot (QD) of 30-35 nm hydrodynamic diameter with varied multivalency from 10 to 75 (e.g., QD(TAT), QD(TAT), QD(TAT), QD(TAT)) and studied the role of multivalency in endocytosis and subcellular trafficking. We found that both low and high multivalent nanoparticles enter into cell predominantly via lipid-raft mediated endocytosis but the higher multivalency of 40 and 75 induces vesicular trapping followed by exocytosis within 12 h. In contrast, lower multivalency of 10 and 20 offers efficient trafficking toward perinuclear region and Golgi apparatus. This work shows the functional role of nanoparticle multivalency in cellular uptake mechanism and importance of lower multivalency for efficient subcellular targeting.
尽管反式激活转录（TAT）肽功能化的纳米颗粒/聚合物/脂质体广泛应用于细胞转染，但颗粒的多价性（每个颗粒连接的肽数量）对细胞摄取机制和亚细胞靶向性能的影响在很大程度上尚未得到探索。在此，我们表明纳米颗粒的多价性控制细胞相互作用、细胞进出机制以及亚细胞靶向性能。我们合成了水动力直径为30 - 35 nm、多价性从10到75不等的TAT肽功能化量子点（QD）（例如QD(TAT)₁₀、QD(TAT)₂₀、QD(TAT)₄₀、QD(TAT)₇₅），并研究了多价性在内吞作用和亚细胞运输中的作用。我们发现，低价和高价多价纳米颗粒主要通过脂筏介导的内吞作用进入细胞，但40和75的较高多价性会诱导囊泡捕获，随后在12小时内发生胞吐作用。相比之下，10和20的较低多价性则能有效地运输到核周区域和高尔基体。这项工作展示了纳米颗粒多价性在细胞摄取机制中的功能作用，以及较低多价性对高效亚细胞靶向的重要性。
