Dong Jiantong, Zhang Ryan Y, Sun Na, Hu Junhui, Smalley Matthew D, Zhou Anqi, Yue Hua, Rothermich Winston, Chen Mengxiang, Chen Jiayuan, Ye Jinglei, Teng Pai-Chi, Qi Dongping, Toretsky Jeffrey A, Tomlinson James S, Li Mengyuan, Weiss Paul S, Jonas Steven J, Federman Noah, Wu Lily, Zhao Meiping, Tseng Hsian-Rong, Zhu Yazhen
California NanoSystems Institute Crump Institute for Molecular Imaging Department of Molecular and Medical Pharmacology University of California Los Angeles (UCLA) 570 Westwood Plaza, Los Angeles, CA 90095, USA.
Department of Molecular and Medical Pharmacology David Geffen School of Medicine UCLA 650 Charles E Young Dr., Los Angeles, CA 90095, USA.
Adv Funct Mater. 2020 Dec 1;30(49). doi: 10.1002/adfm.202003237. Epub 2020 Sep 13.
Tumor-derived extracellular vesicles (EVs) play essential roles in intercellular communication during tumor growth and metastatic evolution. Currently, little is known about the possible roles of tumor-derived EVs in sarcoma because the lack of specific surface markers makes it technically challenging to purify sarcoma-derived EVs. In this study, a specific purification system is developed for Ewing sarcoma (ES)-derived EVs by coupling covalent chemistry-mediated EV capture/ release within a nanostructure-embedded microchip. The purification platform-ES-EV Click Chip-takes advantage of specific anti-LINGO-1 recognition and sensitive click chemistry-mediated EV capture, followed by disulfide cleavage-driven EV release. Since the device is capable of specific and efficient purification of intact ES EVs with high purity, ES-EV Click Chip is ideal for conducting downstream functional studies of ES EVs. Absolute quantification of the molecular hallmark of ES (i.e., EWS rearrangements) using reverse transcription Droplet Digital PCR enables specific quantification of ES EVs. The purified ES EVs can be internalized by recipient cells and transfer their mRNA cargoes, exhibiting their biological intactness and potential role as biological shuttles in intercellular communication.
肿瘤衍生的细胞外囊泡（EVs）在肿瘤生长和转移演变过程中的细胞间通讯中发挥着重要作用。目前，关于肿瘤衍生的EVs在肉瘤中的可能作用知之甚少，因为缺乏特异性表面标志物使得纯化肉瘤衍生的EVs在技术上具有挑战性。在本研究中，通过在嵌入纳米结构的微芯片内结合共价化学介导的EV捕获/释放，开发了一种用于尤因肉瘤（ES）衍生的EVs的特异性纯化系统。该纯化平台——ES-EV点击芯片利用特异性抗LINGO-1识别和灵敏的点击化学介导的EV捕获，随后通过二硫键裂解驱动EV释放。由于该装置能够特异性、高效地纯化高纯度的完整ES EVs，ES-EV点击芯片非常适合进行ES EVs的下游功能研究。使用逆转录液滴数字PCR对ES的分子标志物（即EWS重排）进行绝对定量，能够对ES EVs进行特异性定量。纯化的ES EVs可被受体细胞内化并转移其mRNA货物，显示出它们的生物学完整性以及作为细胞间通讯中生物穿梭体的潜在作用。
