Real‐Time Dissection of the Transportation and miRNA‐Release Dynamics of Small Extracellular Vesicles

Abstract

Extracellular vesicles (EVs) are cell‐derived membrane‐enclosed structures that deliver biomolecules for intercellular communication. Developing visualization methods to elucidate the spatiotemporal dynamics of EVs’ behaviors will facilitate their understanding and translation. With a quantum dot (QD) labeling strategy, a single particle tracking (SPT) platform is proposed here for dissecting the dynamic behaviors of EVs. The interplays between tumor cell‐derived small EVs (T‐sEVs) and endothelial cells (ECs) are specifically investigated based on this platform. It is revealed that, following a clathrin‐mediated endocytosis by ECs, T‐sEVs are transported to the perinuclear region in a typical three‐stage pattern. Importantly, T‐sEVs frequently interact with and finally enter lysosomes, followed by quick release of their carried miRNAs. This study, for the first time, reports the entire process and detailed dynamics of T‐sEV transportation and cargo‐release in ECs, leading to better understanding of their proangiogenic functions. Additionally, the QD‐based SPT technique will help uncover more secrets of sEV‐mediated cell–cell communication.

Hypoxic‐preconditioned mesenchymal stem cell‐derived small extracellular vesicles promote the recovery of spinal cord injury by affecting the phenotype of astrocytes through the miR‐21/JAK2/STAT3 pathway

“…Recent studies have shown that content miRNA plays an important role in the function of EV. 63 , 64 In the animal model of myocardial infarction, EV exerts anti‐apoptotic effects and promotes cardiac tissue and functional recovery by delivering miR‐21 to endothelial cells and cardiomyocytes. 65 In the rat SCI model, EV loaded with miR‐29 transmits miR‐29 to nerve cell, promoting motor and sensory function repair.…”

Extracellular vesicles in tumor angiogenesis and resistance to anti‐angiogenic therapy

“…We recently found that following clathrin‐mediated endocytosis by ECs, T‐sEVs are transported to the perinuclear region in a typical three‐stage pattern. Importantly, T‐sEVs frequently interact with and finally enter lysosomes, followed by a quick release of their carried miRNAs 12 . The proliferation, migration, and tube formation of ECs are finely controlled by T‐EVs via multiple pathways.…”

Real-Time Dissection of the Exosome Pathway for Influenza Virus Infection

“…Quantum dots (QDs) are promising fluorescent probes due to their advantages of broad excitation, narrow emission, and stable luminescence and are widely used for fluorescent labeling of biomaterials . QD-based single-particle tracking (QSPT) has emerged as a robust technique for investigating dynamic information on biomolecules and has been extensively used to investigate the dynamic mechanisms of virus infection in live cells. − QDs with different emission wavelengths are used to label distinct viral components that help to obtain dynamic information on the infection pathway and genome release of labeled viruses by SPT. − However, although our group has previously used QDs to track exosomes by labeling the membrane structure, , a simultaneous dual-color labeling strategy for the internal and external components of exosomes with QDs is still lacking, which is of great importance for the real-time tracking of individual exosomes and elucidating the role of the exosome during virus infection.…”

“…19−25 QDs with different emission wavelengths are used to label distinct viral components that help to obtain dynamic information on the infection pathway and genome release of labeled viruses by SPT. 26−33 However, although our group has previously used QDs to track exosomes by labeling the membrane structure, 34,35 a simultaneous dual-color labeling strategy for the internal and external components of exosomes with QDs is still lacking, which is of great importance for the real-time tracking of individual exosomes and elucidating the role of the exosome during virus infection.…”