Gold nanoparticle based double-labeling of melanoma extracellular vesicles to determine the specificity of uptake by cells and preferential accumulation in small metastatic lung tumors
Gold nanoparticle based double-labeling of melanoma extracellular vesicles to determine the specificity of uptake by cells and preferential accumulation in small metastatic lung tumors
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DOI：
10.1186/s12951-020-0573-0
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发表时间：
2020-01-23
影响因子：
10.2
通讯作者：
Kogan, Marcelo J.
中科院分区：
文献类型：
--
作者：
Lara, Pablo;Palma-Florez, Sujey;Kogan, Marcelo J.
细胞外囊泡（EV）在靶向治疗方面显示出巨大的潜力，因为它们具有通过生物屏障的天然能力，并且根据其来源，可以优先在包括肿瘤在内的特定部位积聚。分析EV靶向特定细胞的潜力仍然具有挑战性，考虑到亲脂性示踪剂与其他蛋白质的非特异性结合，深部组织成像的荧光限制以及外部标记策略对其天然向性的影响。在这项工作中，我们通过荧光分析和定量金标记测量确定了B16 F10-EV对癌细胞和转移性肿瘤的细胞类型特异性向性。等离子体金纳米颗粒的表面功能化用于促进EV的间接标记，而不影响尺寸分布、多分散性、表面电荷、蛋白质标记物、细胞摄取或体内生物分布。将具有金和荧光染料的双标记EV注射到发展转移性肺结节的动物中，并通过荧光/计算机断层扫描成像、定量中子活化分析和金增强光学显微镜进行分析。结果与结肠腺癌、巨噬细胞和肾细胞来源的EV相比，我们确定B16 F10细胞优先摄取其自身的EV。此外，与其他器官相比，我们能够检测到B16 F10 EV在位于肺中的小转移性肿瘤中的优先积累，以及通过组织学分析它们在肿瘤血管，肺泡和肿瘤结节之间的精确分布。最后，我们观察到肿瘤EV可用作有效载体以增加金纳米颗粒向转移性结节的递送。结论我们的研究结果为研究EV在小鼠体内的分布和相互作用提供了一个有价值的工具，并为利用恶性EV的天然特性提高金纳米颗粒对癌细胞和转移性结节的靶向性提供了一种新的策略。
Background Extracellular vesicles (EVs) have shown great potential for targeted therapy, as they have a natural ability to pass through biological barriers and, depending on their origin, can preferentially accumulate at defined sites, including tumors. Analyzing the potential of EVs to target specific cells remains challenging, considering the unspecific binding of lipophilic tracers to other proteins, the limitations of fluorescence for deep tissue imaging and the effect of external labeling strategies on their natural tropism. In this work, we determined the cell-type specific tropism of B16F10-EVs towards cancer cell and metastatic tumors by using fluorescence analysis and quantitative gold labeling measurements. Surface functionalization of plasmonic gold nanoparticles was used to promote indirect labeling of EVs without affecting size distribution, polydispersity, surface charge, protein markers, cell uptake or in vivo biodistribution. Double-labeled EVs with gold and fluorescent dyes were injected into animals developing metastatic lung nodules and analyzed by fluorescence/computer tomography imaging, quantitative neutron activation analysis and gold-enhanced optical microscopy. Results We determined that B16F10 cells preferentially take up their own EVs, when compared with colon adenocarcinoma, macrophage and kidney cell-derived EVs. In addition, we were able to detect the preferential accumulation of B16F10 EVs in small metastatic tumors located in lungs when compared with the rest of the organs, as well as their precise distribution between tumor vessels, alveolus and tumor nodules by histological analysis. Finally, we observed that tumor EVs can be used as effective vectors to increase gold nanoparticle delivery towards metastatic nodules. Conclusions Our findings provide a valuable tool to study the distribution and interaction of EVs in mice and a novel strategy to improve the targeting of gold nanoparticles to cancer cells and metastatic nodules by using the natural properties of malignant EVs.