Coupling Nanostructured Microchips with Covalent Chemistry Enables Purification of Sarcoma‐Derived Extracellular Vesicles for Downstream Functional Studies. (13th September 2020) - Dong, Jiantong
Abstract: 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. Abstract : A novel extracellular vesicle (EV) purification system is developed to realize the specific capture and recovery of Ewing sarcoma (ES)‐derived EVs. The device exploits specific anti‐LINGO‐1 identification of ES EVs, sensitive click chemistry‐mediated ES‐EV capture, and disulfide cleavage‐driven ES‐EV release within a nanostructure‐embedded microfluidic platform, maintaining the high viability and cargo contents of ES EVs for conducting downstream functional studies.