Quantitative characterization of extracellular vesicle uptake and content delivery within mammalian cells

Emeline Bonsergent, Eleonora Grisard, Julian Buchrieser, Olivier Schwartz, Clotilde Théry and Grégory Lavieu ()
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Emeline Bonsergent: Institut Curie, PSL Research University, INSERM U932
Eleonora Grisard: Institut Curie, PSL Research University, INSERM U932
Julian Buchrieser: Institut Pasteur, Virus and Immunity Unit, Department of Virology, CNRS, UMR 3569
Olivier Schwartz: Institut Pasteur, Virus and Immunity Unit, Department of Virology, CNRS, UMR 3569
Clotilde Théry: Institut Curie, PSL Research University, INSERM U932
Grégory Lavieu: Institut Curie, PSL Research University, INSERM U932

Nature Communications, 2021, vol. 12, issue 1, 1-11

Abstract: Abstract Extracellular vesicles (EVs), including exosomes, are thought to mediate intercellular communication through the transfer of cargoes from donor to acceptor cells. Occurrence of EV-content delivery within acceptor cells has not been unambiguously demonstrated, let alone quantified, and remains debated. Here, we developed a cell-based assay in which EVs containing luciferase- or fluorescent-protein tagged cytosolic cargoes are loaded on unlabeled acceptor cells. Results from dose-responses, kinetics, and temperature-block experiments suggest that EV uptake is a low yield process (~1% spontaneous rate at 1 h). Further characterization of this limited EV uptake, through fractionation of membranes and cytosol, revealed cytosolic release (~30% of the uptaken EVs) in acceptor cells. This release is inhibited by bafilomycin A1 and overexpression of IFITM proteins, which prevent virus entry and fusion. Our results show that EV content release requires endosomal acidification and suggest the involvement of membrane fusion.

Date: 2021
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DOI: 10.1038/s41467-021-22126-y

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