Shedding light on the cell biology of extracellular vesicles

Nature Reviews Molecular Cell Biology - Tập 19 Số 4 - Trang 213-228 - 2018
Guillaume van Niel1, Gisela D’angelo2,3, Graça Raposo3
1IPNP - U1266 Inserm - Paris Descartes - Institut de psychiatrie et neurosciences de Paris (102-108 Rue de la Santé 75014 PARIS - France)
2Biologie Cellulaire et Cancer (26 Rue d'Ulm 75248 PARIS CEDEX 05 - France)
3CDC - Compartimentation et dynamique cellulaires (26 Rue d'Ulm 75248 PARIS CEDEX 05 - France)

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Schorey, J. S., Cheng, Y., Singh, P. P. & Smith, V. L. Exosomes and other extracellular vesicles in host-pathogen interactions. EMBO Rep. 16, 24–43 (2015).

Deatherage, B. L. & Cookson, B. T. Membrane vesicle release in bacteria, eukaryotes, and archaea: a conserved yet underappreciated aspect of microbial life. Infect. Immun. 80, 1948–1957 (2012).

Robinson, D. G., Ding, Y. & Jiang, L. Unconventional protein secretion in plants: a critical assessment. Protoplasma 253, 31–43 (2016).

Johnstone, R. M., Adam, M., Hammond, J. R., Orr, L. & Turbide, C. Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles (exosomes). J. Biol. Chem. 262, 9412–9420 (1987). This is the first report of exosomes as intraluminal vesicles of multivesicular endosomes that are secreted upon fusion of these endosomes with the plasma membrane, coining the term 'exosomes'.

Colombo, M., Raposo, G. & Thery, C. Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu. Rev. Cell Dev. Biol. 30, 255–289 (2014).

Lo Cicero, A., Stahl, P. D. & Raposo, G. Extracellular vesicles shuffling intercellular messages: for good or for bad. Curr. Opin. Cell Biol. 35, 69–77 (2015).

Yanez-Mo, M. et al. Biological properties of extracellular vesicles and their physiological functions. J. Extracell. Vesicles 4, 27066 (2015).

Harding, C., Heuser, J. & Stahl, P. Receptor-mediated endocytosis of transferrin and recycling of the transferrin receptor in rat reticulocytes. J. Cell Biol. 97, 329–339 (1983).

Thery, C., Amigorena, S., Raposo, G. & Clayton, A. Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr. Protoc. Cell Biol. http://dx.doi.org/10.1002/0471143030.cb0322s30 (2006). This is the first exhaustive and detailed protocol providing isolation and characterization procedures of exosomes present in cell culture supernatants and biological fluids.

Raposo, G. & Stoorvogel, W. Extracellular vesicles: exosomes, microvesicles, and friends. J. Cell Biol. 200, 373–383 (2013).

Wasmuth, E. V., Januszyk, K. & Lima, C. D. Structure of an Rrp6-RNA exosome complex bound to poly(A) RNA. Nature 511, 435–439 (2014).

Trams, E. G., Lauter, C. J., Salem, N. Jr & Heine, U. Exfoliation of membrane ecto-enzymes in the form of micro-vesicles. Biochim. Biophys. Acta 645, 63–70 (1981).

Harding, C., Heuser, J. & Stahl, P. Endocytosis and intracellular processing of transferrin and colloidal-gold transferrin in rat reticulocytes: demonstration of a pathway for receptor shedding. Eur. J. Cell Biol. 35, 256–263 (1984).

Pan, B. T., Teng, K., Wu, C., Adam, M. & Johnstone, R. M. Electron microscopic evidence for externalization of the transferrin receptor in vesicular form in sheep reticulocytes. J. Cell Biol. 101, 942–948 (1985).

Raposo, G. et al. B lymphocytes secrete antigen-presenting vesicles. J. Exp. Med. 183, 1161–1172 (1996). Exploiting electron microscopy, biochemistry and functional assays, this manuscript shows for the first time that antigen-presenting cells secrete exosomes able to stimulate T cell proliferation.

Zitvogel, L. et al. Eradication of established murine tumors using a novel cell-free vaccine: dendritic cell-derived exosomes. Nat. Med. 4, 594–600 (1998). This study using tumour-bearing mice reports that exosomes secreted by dendritic cells induce antitumoural immune responses in vivo.

Wolf, P. The nature and significance of platelet products in human plasma. Br. J. Haematol. 13, 269–288 (1967).

Stein, J. M. & Luzio, J. P. Ectocytosis caused by sublytic autologous complement attack on human neutrophils. The sorting of endogenous plasma-membrane proteins and lipids into shed vesicles. Biochem. J. 274, 381–386 (1991).

Sims, P. J., Faioni, E. M., Wiedmer, T. & Shattil, S. J. Complement proteins C5b-9 cause release of membrane vesicles from the platelet surface that are enriched in the membrane receptor for coagulation factor Va and express prothrombinase activity. J. Biol. Chem. 263, 18205–18212 (1988).

Satta, N. et al. Monocyte vesiculation is a possible mechanism for dissemination of membrane-associated procoagulant activities and adhesion molecules after stimulation by lipopolysaccharide. J. Immunol. 153, 3245–3255 (1994).

Al-Nedawi, K. et al. Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles derived from tumour cells. Nat. Cell Biol. 10, 619–624 (2008).

Tricario, C., Clancy, J. & De Souza-Schorey, C. Biology and biogenesis of shed microvesicles. Small GTPases 1–13 (2016).

Willms, E. et al. Cells release subpopulations of exosomes with distinct molecular and biological properties. Sci. Rep. 6, 22519 (2016).

Ma, L. et al. Discovery of the migrasome, an organelle mediating release of cytoplasmic contents during cell migration. Cell Res. 25, 24–38 (2015).

Nabhan, J. F., Hu, R., Oh, R. S., Cohen, S. N. & Lu, Q. Formation and release of arrestin domain-containing protein 1-mediated microvesicles (ARMMs) at plasma membrane by recruitment of TSG101 protein. Proc. Natl Acad. Sci. USA 109, 4146–4151 (2012).

Gould, S. J. & Raposo, G. As we wait: coping with an imperfect nomenclature for extracellular vesicles. J. Extracell. Vesicles 2, 20398 (2013).

Kowal, J. et al. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proc. Natl Acad. Sci. USA 113, E968–E977 (2016).

Booth, A. M. et al. Exosomes and HIV Gag bud from endosome-like domains of the T cell plasma membrane. J. Cell Biol. 172, 923–935 (2006).

Gerber, P. P. et al. Rab27a controls HIV-1 assembly by regulating plasma membrane levels of phosphatidylinositol 4,5-bisphosphate. J. Cell Biol. 209, 435–452 (2015).

Kalra, H., Drummen, G. P. & Mathivanan, S. Focus on extracellular vesicles: introducing the next small big thing. Int. J. Mol. Sci. 17, 170 (2016).

Minciacchi, V. R., Freeman, M. R. & Di Vizio, D. Extracellular vesicles in cancer: exosomes, microvesicles and the emerging role of large oncosomes. Semin. Cell Dev. Biol. 40, 41–51 (2015).

Ostrowski, M. et al. Rab27a and Rab27b control different steps of the exosome secretion pathway. Nat. Cell Biol. 12, 19–30 (2010). This study uses a medium-throughput RNA-interference screen for RAB GTPases to reveal the involvement of RAB27 in exosome secretion, providing further evidence that exosomes derive from secretory multivesicular endosomes.

Berson, J. F., Harper, D., Tenza, D., Raposo, G. & Marks, M. S. Pmel17 initiates premelanosome morphogenesis within multivesicular bodies. Mol. Biol. Cell 12, 3451–3464 (2001).

Klumperman, J. & Raposo, G. The complex ultrastructure of the endolysosomal system. Cold Spring Harbor Perspect. Biol. 6, a016857 (2014).

Vidal, M., Mangeat, P. & Hoekstra, D. Aggregation reroutes molecules from a recycling to a shedding pathway during reticulocyte maturation. J. Cell Sci. 110, 1867–1877 (1997).

Zimmermann, P. et al. Syndecan recycling [corrected] is controlled by syntenin-PIP2 interaction and Arf6. Dev. Cell 9, 377–388 (2005).

Baietti, M. F. et al. Syndecan-syntenin-ALIX regulates the biogenesis of exosomes. Nat. Cell Biol. 14, 677–685 (2012). This study reveals the involvement of a particular machinery required for exosome biogenesis and signalling through the interaction of ESCRTs, ALIX and syndecans.

D'Souza-Schorey, C. & Chavrier, P. ARF proteins: roles in membrane traffic and beyond. Nat. Rev. Mol. Cell Biol. 7, 347–358 (2006).

Muralidharan-Chari, V. et al. ARF6-regulated shedding of tumor cell-derived plasma membrane microvesicles. Curr. Biol. 19, 1875–1885 (2009). This is the first report showing a mechanism for microvesicle release through the involvement of ARF6 and cytoskeletal rearrangements. It also shows that microvesicles but not exosomes carry metalloproteases able to digest the extracellular matrix.

Wang, T. et al. Hypoxia-inducible factors and RAB22A mediate formation of microvesicles that stimulate breast cancer invasion and metastasis. Proc. Natl Acad. Sci. USA 111, E3234–E3242 (2014).

Hurley, J. H. ESCRT complexes and the biogenesis of multivesicular bodies. Curr. Opin. Cell Biol. 20, 4–11 (2008).

Tamai, K. et al. Exosome secretion of dendritic cells is regulated by Hrs, an ESCRT-0 protein. Biochem. Biophys. Res. Commun. 399, 384–390 (2010).

Colombo, M. et al. Analysis of ESCRT functions in exosome biogenesis, composition and secretion highlights the heterogeneity of extracellular vesicles. J. Cell Sci. 126, 5553–5565 (2013). This study uses a high-throughput RNA-interference screen targeting ESCRT subunits and accessory proteins to reveal a role for selected ESCRT components in modulation of exosome secretion and composition.

Stuffers, S., Sem Wegner, C., Stenmark, H. & Brech, A. Multivesicular endosome biogenesis in the absence of ESCRTs. Traffic 10, 925–937 (2009).

Trajkovic, K. et al. Ceramide triggers budding of exosome vesicles into multivesicular endosomes. Science 319, 1244–1247 (2008).

Goni, F. M. & Alonso, A. Effects of ceramide and other simple sphingolipids on membrane lateral structure. Biochim. Biophys. Acta 1788, 169–177 (2009).

Kajimoto, T., Okada, T., Miya, S., Zhang, L. & Nakamura, S. Ongoing activation of sphingosine 1-phosphate receptors mediates maturation of exosomal multivesicular endosomes. Nat. Commun. 4, 2712 (2013).

Theos, A. C. et al. A lumenal domain-dependent pathway for sorting to intralumenal vesicles of multivesicular endosomes involved in organelle morphogenesis. Dev. Cell 10, 343–354 (2006).

van Niel, G. et al. The tetraspanin CD63 regulates ESCRT-independent and -dependent endosomal sorting during melanogenesis. Dev. Cell 21, 708–721 (2011).

van Niel, G. et al. Apolipoprotein E regulates amyloid formation within endosomes of pigment cells. Cell Rep. 13, 43–51 (2015).

Gauthier, S. A. et al. Enhanced exosome secretion in Down syndrome brain - a protective mechanism to alleviate neuronal endosomal abnormalities. Acta Neuropathol. Commun. 5, 65 (2017).

Buschow, S. I. et al. MHC II in dendritic cells is targeted to lysosomes or T cell-induced exosomes via distinct multivesicular body pathways. Traffic 10, 1528–1542 (2009). This study reports that upon interaction with T cells, dendritic cells target MHC II molecules towards exosome secretion by using a multivesicular body pathway distinct from those employed by dendritic cells to target MHC II molecules towards lysosomal degradation.

Chairoungdua, A., Smith, D. L., Pochard, P., Hull, M. & Caplan, M. J. Exosome release of beta-catenin: a novel mechanism that antagonizes Wnt signaling. J. Cell Biol. 190, 1079–1091 (2010).

Charrin, S., Jouannet, S., Boucheix, C. & Rubinstein, E. Tetraspanins at a glance. J. Cell Sci. 127, 3641–3648 (2014).

Zimmerman, B. et al. Crystal structure of a full-length human tetraspanin reveals a cholesterol-binding pocket. Cell 167, 1041–1051.e11 (2016).

Odintsova, E. et al. Metastasis suppressor tetraspanin CD82/KAI1 regulates ubiquitylation of epidermal growth factor receptor. J. Biol. Chem. 288, 26323–26334 (2013).

Thery, C. et al. Proteomic analysis of dendritic cell-derived exosomes: a secreted subcellular compartment distinct from apoptotic vesicles. J. Immunol. 166, 7309–7318 (2001).

Geminard, C., De Gassart, A., Blanc, L. & Vidal, M. Degradation of AP2 during reticulocyte maturation enhances binding of hsc70 and Alix to a common site on TfR for sorting into exosomes. Traffic 5, 181–193 (2004).

de Gassart, A., Geminard, C., Fevrier, B., Raposo, G. & Vidal, M. Lipid raft-associated protein sorting in exosomes. Blood 102, 4336–4344 (2003).

Buschow, S. I., Liefhebber, J. M., Wubbolts, R. & Stoorvogel, W. Exosomes contain ubiquitinated proteins. Blood Cells Mol. Dis. 35, 398–403 (2005).

Luhtala, N., Aslanian, A., Yates, J. R. 3rd & Hunter, T. Secreted glioblastoma nanovesicles contain intracellular signaling proteins and active Ras incorporated in a farnesylation-dependent manner. J. Biol. Chem. 292, 611–628 (2017).

Valadi, H. et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat. Cell Biol. 9, 654–659 (2007). This article demonstrates that exosomes contain mRNAs and miRNAs and mediate their transfer between cells.

Nolte-'t Hoen, E. N. et al. Deep sequencing of RNA from immune cell-derived vesicles uncovers the selective incorporation of small non-coding RNA biotypes with potential regulatory functions. Nucleic Acids Res. 40, 9272–9285 (2012).

Thakur, B. K. et al. Double-stranded DNA in exosomes: a novel biomarker in cancer detection. Cell Res. 24, 766–769 (2014).

Kahlert, C. et al. Identification of double-stranded genomic DNA spanning all chromosomes with mutated KRAS and p53 DNA in the serum exosomes of patients with pancreatic cancer. J. Biol. Chem. 289, 3869–3875 (2014).

Villarroya-Beltri, C. et al. Sumoylated hnRNPA2B1 controls the sorting of miRNAs into exosomes through binding to specific motifs. Nat. Commun. 4, 2980 (2013). This study shows that a ribonuclear sumoylated protein binds an RNA motif to promote the targeting of miRNA to exosomes.

Mateescu, B. et al. Obstacles and opportunities in the functional analysis of extracellular vesicle RNA - an ISEV position paper. J. Extracell. Vesicles 6, 1286095 (2017).

Irion, U. & St Johnston, D. bicoid RNA localization requires specific binding of an endosomal sorting complex. Nature 445, 554–558 (2007).

Perez-Hernandez, D. et al. The intracellular interactome of tetraspanin-enriched microdomains reveals their function as sorting machineries toward exosomes. J. Biol. Chem. 288, 11649–11661 (2013).

Gibbings, D. J., Ciaudo, C., Erhardt, M. & Voinnet, O. Multivesicular bodies associate with components of miRNA effector complexes and modulate miRNA activity. Nature Cell Biol. 11, 1143–1149 (2009).

McKenzie, A. J. et al. KRAS–MEK signaling controls Ago2 sorting into exosomes. Cell Rep. 15, 978–987 (2016).

Teng, Y. et al. MVP-mediated exosomal sorting of miR-193a promotes colon cancer progression. Nat. Commun. 8, 14448 (2017).

Shurtleff, M. J., Temoche-Diaz, M. M., Karfilis, K. V., Ri, S. & Schekman, R. Y-Box protein 1 is required to sort microRNAs into exosomes in cells and in a cell-free reaction. eLife 5, e19276 (2016).

Carayon, K. et al. Proteolipidic composition of exosomes changes during reticulocyte maturation. J. Biol. Chem. 286, 34426–34439 (2011).

Segura, E., Amigorena, S. & Thery, C. Mature dendritic cells secrete exosomes with strong ability to induce antigen-specific effector immune responses. Blood Cells Mol. Dis. 35, 89–93 (2005).

Mobius, W. et al. Immunoelectron microscopic localization of cholesterol using biotinylated and non-cytolytic perfringolysin O. J. Histochem. Cytochem. 50, 43–55 (2002).

van Niel, G. et al. Dendritic cells regulate exposure of MHC class II at their plasma membrane by oligoubiquitination. Immunity 25, 885–894 (2006).

Edgar, J. R., Eden, E. R. & Futter, C. E. Hrs- and CD63-dependent competing mechanisms make different sized endosomal intraluminal vesicles. Traffic 15, 197–211 (2014).

Hristov, M., Erl, W., Linder, S. & Weber, P. C. Apoptotic bodies from endothelial cells enhance the number and initiate the differentiation of human endothelial progenitor cells in vitro. Blood 104, 2761–2766 (2004).

Piccin, A., Murphy, W. G. & Smith, O. P. Circulating microparticles: pathophysiology and clinical implications. Blood Rev. 21, 157–171 (2007).

Jimenez, J. J. et al. Endothelial cells release phenotypically and quantitatively distinct microparticles in activation and apoptosis. Thromb. Res. 109, 175–180 (2003).

Connor, D. E., Exner, T., Ma, D. D. & Joseph, J. E. The majority of circulating platelet-derived microparticles fail to bind annexin V, lack phospholipid-dependent procoagulant activity and demonstrate greater expression of glycoprotein Ib. Thromb. Haemost. 103, 1044–1052 (2010).

Del Conde, I., Shrimpton, C. N., Thiagarajan, P. & Lopez, J. A. Tissue-factor-bearing microvesicles arise from lipid rafts and fuse with activated platelets to initiate coagulation. Blood 106, 1604–1611 (2005).

Li, B., Antonyak, M. A., Zhang, J. & Cerione, R. A. RhoA triggers a specific signaling pathway that generates transforming microvesicles in cancer cells. Oncogene 31, 4740–4749 (2012).

McConnell, R. E. et al. The enterocyte microvillus is a vesicle-generating organelle. J. Cell Biol. 185, 1285–1298 (2009).

Warburg, O. On respiratory impairment in cancer cells. Science 124, 269–270 (1956).

Wilson, K. F., Erickson, J. W., Antonyak, M. A. & Cerione, R. A. Rho GTPases and their roles in cancer metabolism. Trends Mol. Med. 19, 74–82 (2013).

Shen, B., Fang, Y., Wu, N. & Gould, S. J. Biogenesis of the posterior pole is mediated by the exosome/microvesicle protein-sorting pathway. J. Biol. Chem. 286, 44162–44176 (2011).

Yang, J. M. & Gould, S. J. The cis-acting signals that target proteins to exosomes and microvesicles. Biochem. Soc. Trans. 41, 277–282 (2013).

Bolukbasi, M. F. et al. miR-1289 and “zipcode”-like sequence enrich mRNAs in microvesicles. Mol. Ther. Nucleic Acids 1, e10 (2012).

Mittelbrunn, M., Vicente-Manzanares, M. & Sanchez-Madrid, F. Organizing polarized delivery of exosomes at synapses. Traffic 16, 327–337 (2015).

Eitan, E., Suire, C., Zhang, S. & Mattson, M. P. Impact of lysosome status on extracellular vesicle content and release. Ageing Res. Rev. 32, 65–74 (2016).

Alvarez-Erviti, L. et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat. Biotechnol. 29, 341–345 (2011).

Villarroya-Beltri, C. et al. ISGylation controls exosome secretion by promoting lysosomal degradation of MVB proteins. Nat. Commun. 7, 13588 (2016).

Edgar, J. R., Manna, P. T., Nishimura, S., Banting, G. & Robinson, M. S. Tetherin is an exosomal tether. eLife 5, e17180 (2016). This study shows that exosomes can be released as clusters that remain attached to each other and anchored to the cell surface by tetherin. It suggests that the tetherin-mediated attachment may have a key role in exosome fate.

Liegeois, S., Benedetto, A., Garnier, J. M., Schwab, Y. & Labouesse, M. The V0-ATPase mediates apical secretion of exosomes containing Hedgehog-related proteins in Caenorhabditis elegans. J. Cell Biol. 173, 949–961 (2006).

Guix, F. X. et al. Tetraspanin 6: a pivotal protein of the multiple vesicular body determining exosome release and lysosomal degradation of amyloid precursor protein fragments. Mol. Neurodegener. 12, 25 (2017).

van Niel, G., Porto-Carreiro, I., Simoes, S. & Raposo, G. Exosomes: a common pathway for a specialized function. J. Biochem. 140, 13–21 (2006).

Papandreou, M. E. & Tavernarakis, N. Autophagy and the endo/exosomal pathways in health and disease. Biotechnol. J. 12, 1600175 (2017).

Dias, M. V. et al. PRNP/prion protein regulates the secretion of exosomes modulating CAV1/caveolin-1-suppressed autophagy. Autophagy 12, 2113–2128 (2016).

Hessvik, N. P. et al. PIKfyve inhibition increases exosome release and induces secretory autophagy. Cell. Mol. Life Sci. 73, 4717–4737 (2016).

Bonifacino, J. S. & Glick, B. S. The mechanisms of vesicle budding and fusion. Cell 116, 153–166 (2004).

Cai, H., Reinisch, K. & Ferro-Novick, S. Coats, tethers, Rabs, and SNAREs work together to mediate the intracellular destination of a transport vesicle. Dev.≈Cell 12, 671–682 (2007).

Mittelbrunn, M. et al. Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells. Nat. Commun. 2, 282 (2011).

Rocha, N. et al. Cholesterol sensor ORP1L contacts the ER protein VAP to control Rab7-RILP-p150 Glued and late endosome positioning. J. Cell Biol. 185, 1209–1225 (2009).

Song, P., Trajkovic, K., Tsunemi, T. & Krainc, D. Parkin modulates endosomal organization and function of the endo-lysosomal pathway. J. Neurosci. 36, 2425–2437 (2016).

Mobius, W. et al. Recycling compartments and the internal vesicles of multivesicular bodies harbor most of the cholesterol found in the endocytic pathway. Traffic 4, 222–231 (2003).

Sinha, S. et al. Cortactin promotes exosome secretion by controlling branched actin dynamics. J. Cell Biol. 214, 197–213 (2016).

Marks, M. S., Heijnen, H. F. & Raposo, G. Lysosome-related organelles: unusual compartments become mainstream. Curr. Opin. Cell Biol. 25, 495–505 (2013).

Hsu, C. et al. Regulation of exosome secretion by Rab35 and its GTPase-activating proteins TBC1D10A-C. J. Cell Biol. 189, 223–232 (2010).

Savina, A., Furlan, M., Vidal, M. & Colombo, M. I. Exosome release is regulated by a calcium-dependent mechanism in K562 cells. J. Biol. Chem. 278, 20083–20090 (2003).

Jahn, R. & Scheller, R. H. SNAREs—engines for membrane fusion. Nat. Rev. Mol. Cell Biol. 7, 631–643 (2006).

Rao, J. & Fitzpatrick, R. E. Use of the Q-switched 755-nm alexandrite laser to treat recalcitrant pigment after depigmentation therapy for vitiligo. Dermatol. Surg. 30, 1043–1045 (2004).

Fader, C. M., Sánchez, D. G., Mestre, M. B. & Colombo, M. I. TI-VAMP/VAMP7 and VAMP3/cellubrevin: two v-SNARE proteins involved in specific steps of the autophagy/multivesicular body pathways. Biochim. Biophys. Acta 1793, 1901–1916 (2009).

Proux-Gillardeaux, V., Raposo, G., Irinopoulou, T. & Galli, T. Expression of the Longin domain of TI-VAMP impairs lysosomal secretion and epithelial cell migration. Biol. Cell 99, 261–271 (2007).

Faure, J. et al. Exosomes are released by cultured cortical neurones. Mol. Cell Neurosci. 31, 642–648 (2006).

Raposo, G. et al. Accumulation of major histocompatibility complex class II molecules in mast cell secretory granules and their release upon degranulation. Mol. Biol. Cell 8, 2631–2645 (1997).

Savina, A., Fader, C. M., Damiani, M. T. & Colombo, M. I. Rab11 promotes docking and fusion of multivesicular bodies in a calcium-dependent manner. Traffic 6, 131–143 (2005).

Puri, N. & Roche, P. A. Mast cells possess distinct secretory granule subsets whose exocytosis is regulated by different SNARE isoforms. Proc. Natl Acad. Sci. USA 105, 2580–2585 (2008).

Wei, Y. et al. Pyruvate kinase type M2 promotes tumour cell exosome release via phosphorylating synaptosome-associated protein 23. Nat. Commun. 8, 14041 (2017).

Gross, J. C., Chaudhary, V., Bartscherer, K. & Boutros, M. Active Wnt proteins are secreted on exosomes. Nat. Cell Biol. 14, 1036–1045 (2012).

Hyenne, V. et al. RAL-1 controls multivesicular body biogenesis and exosome secretion. J. Cell Biol. 211, 27–37 (2015).

Koles, K. & Budnik, V. Exosomes go with the Wnt. Cell Logist 2, 169–173 (2012).

Matsumoto, A. et al. Accelerated growth of B16BL6 tumor in mice through efficient uptake of their own exosomes by B16BL6 cells. Cancer Sci. 108, 1803–1810 (2017).

D'Souza-Schorey, C. & Clancy, J. W. Tumor-derived microvesicles: shedding light on novel microenvironment modulators and prospective cancer biomarkers. Genes Dev. 26, 1287–1299 (2012).

Sedgwick, A. E., Clancy, J. W., Olivia Balmert, M. & D'Souza-Schorey, C. Extracellular microvesicles and invadopodia mediate non-overlapping modes of tumor cell invasion. Sci. Rep. 5, 14748 (2015).

Schlienger, S., Campbell, S. & Claing, A. ARF1 regulates the Rho/MLC pathway to control EGF-dependent breast cancer cell invasion. Mol. Biol. Cell 25, 17–29 (2014).

Wehman, A. M., Poggioli, C., Schweinsberg, P., Grant, B. D. & Nance, J. The P4-ATPase TAT-5 inhibits the budding of extracellular vesicles in C. elegans embryos. Curr. Biol. 21, 1951–1959 (2011).

Taverna, S. et al. Shedding of membrane vesicles mediates fibroblast growth factor-2 release from cells. J. Biol. Chem. 278, 51911–51919 (2003).

Cocucci, E., Racchetti, G. & Meldolesi, J. Shedding microvesicles: artefacts no more. Trends Cell Biol. 19, 43–51 (2009).

Bianco, F. et al. Astrocyte-derived ATP induces vesicle shedding and IL-1 beta release from microglia. J. Immunol. 174, 7268–7277 (2005).

Thomas, L. M. & Salter, R. D. Activation of macrophages by P2X7-induced microvesicles from myeloid cells is mediated by phospholipids and is partially dependent on TLR4. J. Immunol. 185, 3740–3749 (2010).

Bianco, F. et al. Acid sphingomyelinase activity triggers microparticle release from glial cells. EMBO J. 28, 1043–1054 (2009).

Mulcahy, L. A., Pink, R. C. & Carter, D. R. Routes and mechanisms of extracellular vesicle uptake. J. Extracell. Vesicles 3, 24641 (2014).

Denzer, K. et al. Follicular dendritic cells carry MHC class II-expressing microvesicles at their surface. J. Immunol. 165, 1259–1265 (2000).

Mallegol, J. et al. T84-intestinal epithelial exosomes bear MHC class II/peptide complexes potentiating antigen presentation by dendritic cells. Gastroenterology 132, 1866–1876 (2007).

Nolte-'t Hoen, E. N., Buschow, S. I., Anderton, S. M., Stoorvogel, W. & Wauben, M. H. Activated T cells recruit exosomes secreted by dendritic cells via LFA-1. Blood 113, 1977–1981 (2009).

Chivet, M. et al. Exosomes secreted by cortical neurons upon glutamatergic synapse activation specifically interact with neurons. J. Extracell. Vesicles 3, 24722 (2014).

Hoshino, A. et al. Tumour exosome integrins determine organotropic metastasis. Nature 527, 329–335 (2015). This study shows for the first time that exosomal integrins have a key role in directing exosomes from cancer cells to particular organs and that this transfer induces metastasis.

Morelli, A. E. et al. Endocytosis, intracellular sorting and processing of exosomes by dendritic cells. Blood 104, 3257–3266 (2004).

Sung, B. H., Ketova, T., Hoshino, D., Zijlstra, A. & Weaver, A. M. Directional cell movement through tissues is controlled by exosome secretion. Nat. Commun. 6, 7164 (2015).

Purushothaman, A. et al. Fibronectin on the surface of myeloma cell-derived exosomes mediates exosome-cell interactions. J. Biol. Chem. 291, 1652–1663 (2016).

Leiss, M., Beckmann, K., Giros, A., Costell, M. & Fassler, R. The role of integrin binding sites in fibronectin matrix assembly in vivo. Curr. Opin. Cell Biol. 20, 502–507 (2008).

Rana, S., Yue, S., Stadel, D. & Zoller, M. Toward tailored exosomes: the exosomal tetraspanin web contributes to target cell selection. Int. J. Biochem. Cell Biol. 44, 1574–1584 (2012).

Nazarenko, I. et al. Cell surface tetraspanin Tspan8 contributes to molecular pathways of exosome-induced endothelial cell activation. Cancer Res. 70, 1668–1678 (2010).

Rana, S., Claas, C., Kretz, C. C., Nazarenko, I. & Zoeller, M. Activation-induced internalization differs for the tetraspanins CD9 and Tspan8: Impact on tumor cell motility. Int. J. Biochem. Cell Biol. 43, 106–119 (2011).

Melo, S. A. et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer. Nature 523, 177–182 (2015).

Bruno, S. et al. Mesenchymal stem cell-derived microvesicles protect against acute tubular injury. J. Am. Soc. Nephrol. 20, 1053–1067 (2009).

Barres, C. et al. Galectin-5 is bound onto the surface of rat reticulocyte exosomes and modulates vesicle uptake by macrophages. Blood 115, 696–705 (2010).

Frey, B. & Gaipl, U. S. The immune functions of phosphatidylserine in membranes of dying cells and microvesicles. Semin. Immunopathol. 33, 497–516 (2011).

Feng, D. et al. Cellular internalization of exosomes occurs through phagocytosis. Traffic 11, 675–687 (2010).

Nakase, I., Kobayashi, N. B., Takatani-Nakase, T. & Yoshida, T. Active macropinocytosis induction by stimulation of epidermal growth factor receptor and oncogenic Ras expression potentiates cellular uptake efficacy of exosomes. Sci. Rep. 5, 10300 (2015).

Tian, T. et al. Exosome uptake through clathrin-mediated endocytosis and macropinocytosis and mediating miR-21 delivery. J. Biol. Chem. 289, 22258–22267 (2014).

Laulagnier, K. et al. Amyloid precursor protein products concentrate in a subset of exosomes specifically endocytosed by neurons. Cell. Mol. Life Sci. http://dx.doi.org/10.1007/s00018-017-2664-0 (2017).

Vargas, A. et al. Syncytin proteins incorporated in placenta exosomes are important for cell uptake and show variation in abundance in serum exosomes from patients with preeclampsia. FASEB J. 28, 3703–3719 (2014).

Kamerkar, S. et al. Exosomes facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer. Nature 546, 498–503 (2017).

Prada, I. et al. A new approach to follow a single extracellular vesicle-cell interaction using optical tweezers. Biotechniques 60, 35–41 (2016).

Heusermann, W. et al. Exosomes surf on filopodia to enter cells at endocytic hot spots, traffic within endosomes, and are targeted to the ER. J. Cell Biol. 213, 173–184 (2016).

Escrevente, C., Keller, S., Altevogt, P. & Costa, J. Interaction and uptake of exosomes by ovarian cancer cells. BMC Cancer 11, 108 (2011).

Chen, W. et al. Nanoscale characterization of carrier dynamic and surface passivation in InGaN/GaN multiple quantum wells on GaN nanorods. ACS Appl. Mater. Interfaces 8, 31887–31893 (2016).

Tian, T., Wang, Y., Wang, H., Zhu, Z. & Xiao, Z. Visualizing of the cellular uptake and intracellular trafficking of exosomes by live-cell microscopy. J. Cell. Biochem. 111, 488–496 (2010).

Bissig, C. & Gruenberg, J. ALIX and the multivesicular endosome: ALIX in Wonderland. Trends Cell Biol. 24, 19–25 (2014).

Antonyak, M. A. et al. Cancer cell-derived microvesicles induce transformation by transferring tissue transglutaminase and fibronectin to recipient cells. Proc. Natl Acad. Sci. USA 108, 4852–4857 (2011).

Desrochers, L. M., Bordeleau, F., Reinhart-King, C. A., Cerione, R. A. & Antonyak, M. A. Microvesicles provide a mechanism for intercellular communication by embryonic stem cells during embryo implantation. Nat. Commun. 7, 11958 (2016).

Zhang, L. & Wrana, J. L. The emerging role of exosomes in Wnt secretion and transport. Curr. Opin. Genet. Dev. 27, 14–19 (2014).

Skog, J. et al. Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nat. Cell Biol. 10, 1470–1476 (2008).

Record, M., Carayon, K., Poirot, M. & Silvente-Poirot, S. Exosomes as new vesicular lipid transporters involved in cell-cell communication and various pathophysiologies. Biochim. Biophys. Acta 1841, 108–120 (2014).

Coleman, B. M. & Hill, A. F. Extracellular vesicles—their role in the packaging and spread of misfolded proteins associated with neurodegenerative diseases. Semin. Cell Dev. Biol. 40, 89–96 (2015).

van Dongen, H. M., Masoumi, N., Witwer, K. W. & Pegtel, D. M. Extracellular vesicles exploit viral entry routes for cargo delivery. Microbiol. Mol. Biol. Rev. 80, 369–386 (2016).

Zhao, H. et al. Tumor microenvironment derived exosomes pleiotropically modulate cancer cell metabolism. eLife 5, e10250 (2016).

Peinado, H. et al. Pre-metastatic niches: organ-specific homes for metastases. Nat. Rev. Cancer 17, 302–317 (2017).

Fais, S. et al. Evidence-based clinical use of nanoscale extracellular vesicles in nanomedicine. ACS Nano 10, 3886–3899 (2016).

Dinkins, M. B., Dasgupta, S., Wang, G., Zhu, G. & Bieberich, E. Exosome reduction in vivo is associated with lower amyloid plaque load in the 5XFAD mouse model of Alzheimer's disease. Neurobiol. Aging 35, 1792–1800 (2014).

Torrano, V. et al. Vesicle-MaNiA: extracellular vesicles in liquid biopsy and cancer. Curr. Opin. Pharmacol. 29, 47–53 (2016).

El Andaloussi, S., Lakhal, S., Mager, I. & Wood, M. J. Exosomes for targeted siRNA delivery across biological barriers. Adv. Drug Deliv. Rev. 65, 391–397 (2013).

Pitt, J. M. et al. Dendritic cell-derived exosomes as immunotherapies in the fight against cancer. J. Immunol. 193, 1006–1011 (2014).

Besse, B. et al. Dendritic cell-derived exosomes as maintenance immunotherapy after first line chemotherapy in NSCLC. Oncoimmunology 5, e1071008 (2016).

Lener, T. et al. Applying extracellular vesicles based therapeutics in clinical trials - an ISEV position paper. J. Extracell. Vesicles 4, 30087 (2015).

Lo Cicero, A. et al. Exosomes released by keratinocytes modulate melanocyte pigmentation. Nat. Commun. 6, 7506 (2015).

Beauvillain, C., Juste, M. O., Dion, S., Pierre, J. & Dimier-Poisson, I. Exosomes are an effective vaccine against congenital toxoplasmosis in mice. Vaccine 27, 1750–1757 (2009).

Ranghino, A. et al. The effects of glomerular and tubular renal progenitors and derived extracellular vesicles on recovery from acute kidney injury. Stem Cell Res. Ther. 8, 24 (2017).

Arslan, F. et al. Mesenchymal stem cell-derived exosomes increase ATP levels, decrease oxidative stress and activate PI3K/Akt pathway to enhance myocardial viability and prevent adverse remodeling after myocardial ischemia/reperfusion injury. Stem Cell Res. 10, 301–312 (2013).

Ophelders, D. R. et al. Mesenchymal stromal cell-derived extracellular vesicles protect the fetal brain after hypoxia-ischemia. Stem Cells Transl Med. 5, 754–763 (2016).

Mizrak, A. et al. Genetically engineered microvesicles carrying suicide mRNA/protein inhibit schwannoma tumor growth. Mol. Ther. 21, 101–108 (2013).

Pan, S., Yang, X., Jia, Y., Li, R. & Zhao, R. Microvesicle-shuttled miR-130b reduces fat deposition n recipient primary cultured porcine adipocytes by inhibiting PPAR-g expression. J. Cell. Physiol. 229, 631–639 (2014).

Coumans, F. A. W. et al. Methodological guidelines to study extracellular vesicles. Circ. Res. 120, 1632–1648 (2017).

Van Deun, J. et al. EV-TRACK: transparent reporting and centralizing knowledge in extracellular vesicle research. Nat. Methods 14, 228–232 (2017).

Takov, K., Yellon, D. M. & Davidson, S. M. Confounding factors in vesicle uptake studies using fluorescent lipophilic membrane dyes. J. Extracell. Vesicles 6, 1388731 (2017).

Hyenne, V., Lefebvre, O. & Goetz, J. G. Going live with tumor exosomes and microvesicles. Cell Adh. Migr. 11, 173–186 (2017).

Lai, C. P., Tannous, B. A. & Breakefield, X. O. Noninvasive in vivo monitoring of extracellular vesicles. Methods Mol. Biol. 1098, 249–258 (2014).

Zomer, A. et al. In vivo imaging reveals extracellular vesicle-mediated phenocopying of metastatic behavior. Cell 161, 1046–1057 (2015). This study uses in vivo imaging to reveal the existence of a functional transfer of mRNA mediated by extracellular vesicles in the context of cancer and metastasis.

Bobrie, A., Colombo, M., Krumeich, S., Raposo, G. & Thery, C. Diverse subpopulations of vesicles secreted by different intracellular mechanisms are present in exosome preparations obtained by differential ultracentrifugation. J. Extracell. Vesicles 1, 18397 (2012).

Roucourt, B., Meeussen, S., Bao, J., Zimmermann, P. & David, G. Heparanase activates the syndecan-syntenin-ALIX exosome pathway. Cell Res. 25, 412–428 (2015).

van Niel, G. et al. Intestinal epithelial cells secrete exosome-like vesicles. Gastroenterology 121, 337–349 (2001).

Tauro, B. J. et al. Two distinct populations of exosomes are released from LIM1863 colon carcinoma cell-derived organoids. Mol. Cell. Proteomics 12, 587–598 (2013).