PEGylated and targeted extracellular vesicles display enhanced cell specificity and circulation time
PEGylated and targeted extracellular vesicles display enhanced cell specificity and circulation time
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DOI：
10.1016/j.jconrel.2016.01.009
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发表时间：
2016-02-28
影响因子：
10.8
通讯作者：
Schiffelers, R. M.
中科院分区：
文献类型：
--
作者：
Kooijmans, S. A. A.;Fliervoet, L. A. L.;Schiffelers, R. M.
细胞外囊泡（EV）因其在细胞之间功能性转移生物货物的能力而越来越被认为是候选药物递送系统。然而，由于缺乏细胞靶向特异性以及外源性 EV 从循环中快速清除，EV 的治疗适用性可能受到限制。为了改善 EV 向肿瘤细胞递送药物的特性，我们开发了一种用与聚乙二醇 (PEG) 缀合的靶向配体修饰 EV 的新方法。将表皮生长因子受体（EGFR）特异性的纳米抗体与磷脂（DMPE）-PEG衍生物缀合以制备纳米抗体-PEG-胶束。当胶束与来自 Neuro2A 细胞或血小板的 EV 混合时，观察到纳米抗体-PEG-脂质向 EV 膜的温度依赖性转移，表明“插入后”机制。该过程不影响 EV 形态、大小分布或蛋白质组成。将 PEG 缀合的对照纳米抗体引入 EV 后，由于 PEG 的屏蔽特性，细胞结合受到损害。然而，当使用 EGFR 特异性纳米抗体时，与 EGFR 过表达肿瘤细胞的特异性结合显着增加。此外，未修饰的 EV 在小鼠静脉注射后 10 分钟内迅速从循环中清除，而用纳米抗体-PEG-脂质修饰的 EV 在注射后 60 分钟以上仍可在血浆中检测到。总之，我们建议将后插入作为一种新技术，赋予分离的 EV 靶向能力，从而避免修改 EV 分泌细胞的要求。重要的是，在 EV 膜中插入配体缀合的 PEG 衍生化磷脂可以提高 EV 的细胞特异性并延长循环时间，从而可能增加 EV 在目标组织中的积累并改善货物输送。 (C) 2015 Elsevier B.V. 保留所有权利。
Extracellular vesicles (EVs) are increasingly being recognized as candidate drug delivery systems due to their ability to functionally transfer biological cargo between cells. However, the therapeutic applicability of EVs may be limited due to a lack of cell-targeting specificity and rapid clearance of exogenous EVs from the circulation. In order to improve EV characteristics for drug delivery to tumor cells, we have developed a novel method for decorating EVs with targeting ligands conjugated to polyethylene glycol (PEG). Nanobodies specific for the epidermal growth factor receptor (EGFR) were conjugated to phospholipid (DMPE)-PEG derivatives to prepare nanobody-PEG-micelles. When micelles were mixed with EVs derived from Neuro2A cells or platelets, a temperature-dependent transfer of nanobody-PEG-lipids to the EV membranes was observed, indicative of a 'post-insertion' mechanism. This process did not affect EV morphology, size distribution, or protein composition. After introduction of PEG-conjugated control nanobodies to EVs, cellular binding was compromised due to the shielding properties of PEG. However, specific binding to EGFR-overexpressing tumor cells was dramatically increased when EGFR-specific nanobodies were employed. Moreover, whereas unmodified EVs were rapidly cleared from the circulation within 10 min after intravenous injection in mice, EVs modified with nanobody-PEG-lipids were still detectable in plasma for longer than 60 min post-injection. In conclusion, we propose post-insertion as a novel technique to confer targeting capacity to isolated EVs, circumventing the requirement to modify EV-secreting cells. Importantly, insertion of ligand-conjugated PEG-derivatized phospholipids in EV membranes equips EVs with improved cell specificity and prolonged circulation times, potentially increasing EV accumulation in targeted tissues and improving cargo delivery. (C) 2015 Elsevier B.V. All rights reserved.