Department of Bio and Brain Engineering, KAIST, Daejeon, 34141, Korea.
Cellex Life Sciences Inc., Daejeon, 34141, Korea.
Nat Commun. 2016 Jul 22;7:12277. doi: 10.1038/ncomms12277.
Nanoparticle-mediated delivery of functional macromolecules is a promising method for treating a variety of human diseases. Among nanoparticles, cell-derived exosomes have recently been highlighted as a new therapeutic strategy for the in vivo delivery of nucleotides and chemical drugs. Here we describe a new tool for intracellular delivery of target proteins, named 'exosomes for protein loading via optically reversible protein-protein interactions' (EXPLORs). By integrating a reversible protein-protein interaction module controlled by blue light with the endogenous process of exosome biogenesis, we are able to successfully load cargo proteins into newly generated exosomes. Treatment with protein-loaded EXPLORs is shown to significantly increase intracellular levels of cargo proteins and their function in recipient cells in vitro and in vivo. These results clearly indicate the potential of EXPLORs as a mechanism for the efficient intracellular transfer of protein-based therapeutics into recipient cells and tissues.
纳米颗粒介导的功能大分子传递是治疗多种人类疾病的有前途的方法。在纳米颗粒中，细胞衍生的外泌体最近作为体内传递核苷酸和化学药物的新治疗策略而受到关注。在这里，我们描述了一种用于靶蛋白细胞内传递的新工具，命名为“通过光可逆蛋白-蛋白相互作用进行蛋白质加载的外泌体”（EXPLORs）。通过将蓝光控制的可逆蛋白-蛋白相互作用模块与外泌体生物发生的内源性过程相结合，我们能够成功地将货物蛋白加载到新生成的外泌体中。研究表明，用负载蛋白的 EXPLORs 处理可显著增加受体细胞中货物蛋白的细胞内水平及其功能，无论是在体外还是体内。这些结果清楚地表明，EXPLORs 作为一种有效的将基于蛋白质的治疗剂递送到受体细胞和组织中的机制具有潜力。
