Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes

Joanna Kowal (1) , Guillaume Arras (2) , Marina Colombo (1) , Mabel Jouve (3, 1) , Jakob Paul Morath (1) , Bjarke Primdal-Bengtson (1) , Florent Dingli (2) , Damarys Loew (2) , Mercedes Tkach (1) , Clotilde Théry (1)

1 U932 - Immunité et cancer
2 Laboratoire de Spectrométrie de Masse Protéomique
3 Institut Curie [Paris]

Extracellular vesicles (EVs) have become the focus of rising interest because of their numerous functions in physiology and pathology. Cells release heterogeneous vesicles of different sizes and intracellular origins, including small EVs formed inside endosomal compartments (i.e., exosomes) and EVs of various sizes budding from the plasma membrane. Specific markers for the analysis and isolation of different EV populations are missing, imposing important limitations to understanding EV functions. Here, EVs from human dendritic cells were first separated by their sedimentation speed, and then either by their behavior upon upward floatation into iodixanol gradients or by immuno-isolation. Extensive quantitative proteomic analysis allowing comparison of the isolated populations showed that several classically used exosome markers, like major histocompatibility complex, flotillin, and heat-shock 70-kDa proteins, are similarly present in all EVs. We identified proteins specifically enriched in small EVs, and define a set of five protein categories displaying different relative abundance in distinct EV populations. We demonstrate the presence of exosomal and nonexosomal subpopulations within small EVs, and propose their differential separation by immuno-isolation using either CD63, CD81, or CD9. Our work thus provides guidelines to define subtypes of EVs for future functional studies.

Mots clés: dendritic cells; ectosomes; exosomes; extracellular vesicles; microvesicles

Domaines: Sciences du Vivant [q-bio]

Joanna Kowal, Guillaume Arras, Marina Colombo, Mabel Jouve, Jakob Paul Morath, et al.. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113 (8), pp.E968-E977. ⟨10.1073/pnas.1521230113⟩. ⟨inserm-02450678⟩