Specificities of exosome versus small ectosome secretion revealed by live intracellular tracking of CD63 and CD9

Mathilde Mathieu, Nathalie Névo, Mabel Jouve, José Ignacio Valenzuela, Mathieu Maurin, Frederik J. Verweij, Roberta Palmulli, Danielle Lankar, Florent Dingli, Damarys Loew, Eric Rubinstein, Gaëlle Boncompain, Franck Perez and Clotilde Théry ()
Additional contact information
Mathilde Mathieu: INSERM U932, Institut Curie Centre de Recherche, PSL Research University
Nathalie Névo: INSERM U932, Institut Curie Centre de Recherche, PSL Research University
Mabel Jouve: CNRS UMR3215, Institut Curie, PSL Research University
José Ignacio Valenzuela: CNRS UMR144, Institut Curie, PSL Research University
Mathieu Maurin: INSERM U932, Institut Curie Centre de Recherche, PSL Research University
Frederik J. Verweij: Institute of Psychiatry and Neuroscience of Paris (IPNP), INSERM U1266
Roberta Palmulli: Université de Paris
Danielle Lankar: INSERM U932, Institut Curie Centre de Recherche, PSL Research University
Florent Dingli: Institut Curie, PSL Research University, Centre de Recherche, Laboratoire de Spectrométrie de Masse Protéomique
Damarys Loew: Institut Curie, PSL Research University, Centre de Recherche, Laboratoire de Spectrométrie de Masse Protéomique
Eric Rubinstein: Sorbonne Université, INSERM, CNRS, Centre d’Immunologie et des Maladies Infectieuses, CIMI-Paris
Gaëlle Boncompain: CNRS UMR144, Institut Curie, PSL Research University
Franck Perez: CNRS UMR144, Institut Curie, PSL Research University
Clotilde Théry: INSERM U932, Institut Curie Centre de Recherche, PSL Research University

Nature Communications, 2021, vol. 12, issue 1, 1-18

Abstract: Abstract Despite their roles in intercellular communications, the different populations of extracellular vesicles (EVs) and their secretion mechanisms are not fully characterized: how and to what extent EVs form as intraluminal vesicles of endocytic compartments (exosomes), or at the plasma membrane (PM) (ectosomes) remains unclear. Here we follow intracellular trafficking of the EV markers CD9 and CD63 from the endoplasmic reticulum to their residency compartment, respectively PM and late endosomes. We observe transient co-localization at both places, before they finally segregate. CD9 and a mutant CD63 stabilized at the PM are more abundantly released in EVs than CD63. Thus, in HeLa cells, ectosomes are more prominent than exosomes. By comparative proteomic analysis and differential response to neutralization of endosomal pH, we identify a few surface proteins likely specific of either exosomes (LAMP1) or ectosomes (BSG, SLC3A2). Our work sets the path for molecular and functional discrimination of exosomes and small ectosomes in any cell type.

Date: 2021
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DOI: 10.1038/s41467-021-24384-2

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