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

Author

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  • Emeline Bonsergent

    (Institut Curie, PSL Research University, INSERM U932
    Université de Paris, INSERM, CNRS UMR 7057)

  • 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
    Université de Paris, INSERM, CNRS UMR 7057)

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.

Suggested Citation

  • Emeline Bonsergent & Eleonora Grisard & Julian Buchrieser & Olivier Schwartz & Clotilde Théry & Grégory Lavieu, 2021. "Quantitative characterization of extracellular vesicle uptake and content delivery within mammalian cells," Nature Communications, Nature, vol. 12(1), pages 1-11, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-22126-y
    DOI: 10.1038/s41467-021-22126-y
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    Cited by:

    1. Chi-Ling Chiang & Yifan Ma & Ya-Chin Hou & Junjie Pan & Sin-Yu Chen & Ming-Hsien Chien & Zhi-Xuan Zhang & Wei-Hsiang Hsu & Xinyu Wang & Jingjing Zhang & Hong Li & Lili Sun & Shannon Fallen & Inyoul Le, 2023. "Dual targeted extracellular vesicles regulate oncogenic genes in advanced pancreatic cancer," Nature Communications, Nature, vol. 14(1), pages 1-18, December.
    2. Tobias Sahr & Pedro Escoll & Christophe Rusniok & Sheryl Bui & Gérard Pehau-Arnaudet & Gregory Lavieu & Carmen Buchrieser, 2022. "Translocated Legionella pneumophila small RNAs mimic eukaryotic microRNAs targeting the host immune response," Nature Communications, Nature, vol. 13(1), pages 1-18, December.

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