Specificities of exosome versus small ectosome secretion revealed by live intracellular tracking of CD63 and CD9
Mathieu, Mathilde; Névo, Nathalie; Jouve, Mabel; Valenzuela, José Ignacio; Maurin, Mathieu; Verweij, Frederik J; Palmulli, Roberta; Lankar, Danielle; Dingli, Florent; Loew, Damarys; Rubinstein, Eric; Boncompain, Gaëlle; Perez, Franck; Théry, Clotilde
(2021) Nature Communications, volume 12, issue 1
(Article)
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
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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.
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Keywords: Cell Communication, Cell Membrane/metabolism, Endosomes/metabolism, Exosomes/metabolism, Extracellular Vesicles/metabolism, Fusion Regulatory Protein 1, Heavy Chain, Gene Knockout Techniques, HeLa Cells, Humans, Membrane Proteins/metabolism, Protein Transport, Proteomics, Tetraspanin 29/metabolism, Tetraspanin 30/metabolism, General Physics and Astronomy, General Chemistry, General Biochemistry,Genetics and Molecular Biology
ISSN: 2041-1723
Publisher: Nature Publishing Group
Note: Funding Information: We thank for fruitful discussions several team members, especially Drs Mercedes Tkach, Eleonora Grisard, Lorena Martin-Jaular, Jason Ecard, and for helpful discussions and tools, Dr Aurélien Dauphin, Institut Curie, Paris, Dr P. Zimmermann, KU Leuven, Belgium and CRCM Marseille, France, Dr G. van Niel, IPN Paris, France, Dr P Benaroch, Institut Curie, Paris, France, Dr Suresh Mathivanan, LaTrobe University, Melbourne, Australia. The H4A3 monoclonal antibody (anti-human LAMP1), developed by August, J.T. and Hildreth, J.E.K. from Johns Hopkins University School of Medicine, was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH, and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242. This work was funded by INSERM, CNRS, Institut Curie, French IdEx and LabEx (ANR-10-INSB-04, ANR-10-IDEX-0001-02 PSL, ANR-10-LABX-0038, ANR-11-LABX-0043, ANR-18-IDEX-0001 Université de Paris), grants from french ANR (ANR-18-CE13-0017-03; ANR-18-CE15-0008-01; ANR-18-CE16-0022-02), INCa (INCA-11548), Fon-dation ARC (PGA1 RF20180206962), FRM (FDT201904007945, EQU201903007925 and DGE20121125630), Cancéropôle Île-de-France (2013-2-EML-02-ICR-1), USA NIDA (DA040385), and a Long-Term EMBO Fellowship (ALTF 607-2015) co-funded by the European Commission FP7 (Marie Curie Actions, LTFCOFUND2013, GA-2013-609409) to J.I.V. We also acknowledge the following platforms of Institut Curie: PICT-IBiSA, member of the France-BioImaging national research infrastructure (ANR-10-INBS-04) for fluorescence and electron microscopy, flow cytometry for assistance in data acquisition, genomic for authentication of HeLa cells by STR analysis. Publisher Copyright: © 2021, The Author(s).
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