Cellular and Molecular Life Sciences

, Volume 73, Issue 24, pp 4717–4737 | Cite as

PIKfyve inhibition increases exosome release and induces secretory autophagy

  • Nina Pettersen Hessvik
  • Anders Øverbye
  • Andreas Brech
  • Maria Lyngaas Torgersen
  • Ida Seim Jakobsen
  • Kirsten Sandvig
  • Alicia LlorenteEmail author
Original Article


Exosomes are vesicles released from cells by fusion of multivesicular bodies (MVBs) with the plasma membrane. This study aimed to investigate whether the phosphoinositide kinase PIKfyve affects this process. Our results show that in PC-3 cells inhibition of PIKfyve by apilimod or depletion by siRNA increased the secretion of the exosomal fraction. Moreover, quantitative electron microscopy analysis showed that cells treated with apilimod contained more MVBs per cell and more intraluminal vesicles per MVB. Interestingly, mass spectrometry analysis revealed a considerable enrichment of autophagy-related proteins (NBR1, p62, LC3, WIPI2) in exosomal fractions released by apilimod-treated cells, a result that was confirmed by immunoblotting. When the exosome preparations were investigated by electron microscopy a small population of p62-labelled electron dense structures was observed together with CD63-containing exosomes. The p62-positive structures were found in less dense fractions than exosomes in density gradients. Inside the cells, p62 and CD63 were found in the same MVB-like organelles. Finally, both the degradation of EGF and long-lived proteins were shown to be reduced by apilimod. In conclusion, inhibition of PIKfyve increases secretion of exosomes and induces secretory autophagy, showing that these pathways are closely linked. We suggest this is due to impaired fusion of lysosomes with both MVBs and autophagosomes, and possibly increased fusion of MVBs with autophagosomes, and that the cells respond by secreting the content of these organelles to maintain cellular homeostasis.


Exosomes Extracellular vesicles PIKfyve Secretory autophagy Exophagy Phosphoinositides 



Cation-independent mannose 6-phosphate receptor


Electron microscopy


Endosomal sorting complexes required for transport proteins


Intraluminal vesicle


Mass spectrometry


Multivesicular body


Nanoparticle tracking analysis








Trans-golgi network



This work was funded by The South-Eastern Norwegian Regional Health Authority, The Norwegian Cancer Society and The Research Council of Norway, and supported by The Research Council of Norway through its Centers of Excellence funding scheme, project number 179571. We thank Anne Engen, Anne Grethe Myrann and Marianne Smestad for their excellent technical assistance and Tore Skotland for valuable and interesting discussions. We also thank the proteomics core facility, especially Bernd Thiede and Christian Koehler, at the University of Oslo.

Supplementary material

18_2016_2309_MOESM1_ESM.pdf (556 kb)
Supplementary material 1 (PDF 556 kb)


  1. 1.
    Pan BT, Teng K, Wu C, Adam M, Johnstone RM (1985) Electron microscopic evidence for externalization of the transferrin receptor in vesicular form in sheep reticulocytes. J Cell Biol 101(3):942–948. doi: 10.1083/jcb.101.3.942 CrossRefPubMedGoogle Scholar
  2. 2.
    Johnstone RM, Adam M, Hammond JR, Orr L, Turbide C (1987) Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles (exosomes). J Biol Chem 262(19):9412–9420PubMedGoogle Scholar
  3. 3.
    Raposo G, Stoorvogel W (2013) Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol 200(4):373–383. doi: 10.1083/jcb.201211138 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Ostenfeld MS, Jeppesen DK, Laurberg JR, Boysen AT, Bramsen JB, Primdal-Bengtson B, Hendrix A, Lamy P, Dagnaes-Hansen F, Rasmussen MH, Bui KH, Fristrup N, Christensen EI, Nordentoft I, Morth JP, Jensen JB, Pedersen JS, Beck M, Theodorescu D, Borre M, Howard KA, Dyrskjot L, Orntoft TF (2014) Cellular disposal of miR23b by RAB27-dependent exosome release is linked to acquisition of metastatic properties. Cancer Res 74(20):5758–5771. doi: 10.1158/0008-5472.can-13-3512 CrossRefPubMedGoogle Scholar
  5. 5.
    Mathivanan S, Ji H, Simpson RJ (2010) Exosomes: extracellular organelles important in intercellular communication. J Proteomics 73(10):1907–1920CrossRefPubMedGoogle Scholar
  6. 6.
    Record M, Carayon K, Poirot M, Silvente-Poirot S (2014) Exosomes as new vesicular lipid transporters involved in cell-cell communication and various pathophysiologies. Biochim Biophys Acta 1841(1):108–120. doi: 10.1016/j.bbalip.2013.10.004 CrossRefPubMedGoogle Scholar
  7. 7.
    Thery C, Boussac M, Veron P, Ricciardi-Castagnoli P, Raposo G, Garin J, Amigorena S (2001) Proteomic analysis of dendritic cell-derived exosomes: a secreted subcellular compartment distinct from apoptotic vesicles. J Immunol 166(12):7309–7318CrossRefPubMedGoogle Scholar
  8. 8.
    Laulagnier K, Motta C, Hamdi S, Roy S, Fauvelle F, Pageaux JF, Kobayashi T, Salles JP, Perret B, Bonnerot C, Record M (2004) Mast cell- and dendritic cell-derived exosomes display a specific lipid composition and an unusual membrane organization. Biochem J 380(Pt 1):161–171. doi: 10.1042/bj20031594 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Llorente A, Skotland T, Sylvanne T, Kauhanen D, Rog T, Orlowski A, Vattulainen I, Ekroos K, Sandvig K (2013) Molecular lipidomics of exosomes released by PC-3 prostate cancer cells. Biochim Biophys Acta 1831(7):1302–1309CrossRefPubMedGoogle Scholar
  10. 10.
    Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO (2007) Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9(6):654–659CrossRefPubMedGoogle Scholar
  11. 11.
    Mittelbrunn M, Gutierrez-Vazquez C, Villarroya-Beltri C, Gonzalez S, Sanchez-Cabo F, Gonzalez MA, Bernad A, Sanchez-Madrid F (2011) Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells. Nat Commun 2:282CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Hessvik NP, Phuyal S, Brech A, Sandvig K, Llorente A (2012) Profiling of microRNAs in exosomes released from PC-3 prostate cancer cells. Biochim Biophys Acta 1819(11–12):1154–1163CrossRefPubMedGoogle Scholar
  13. 13.
    Ronquist KG, Ronquist G, Carlsson L, Larsson A (2009) Human prostasomes contain chromosomal DNA. Prostate 69(7):737–743. doi: 10.1002/pros.20921 CrossRefPubMedGoogle Scholar
  14. 14.
    Lazaro-Ibanez E, Sanz-Garcia A, Visakorpi T, Escobedo-Lucea C, Siljander P, Ayuso-Sacido A, Yliperttula M (2014) Different gDNA content in the subpopulations of prostate cancer extracellular vesicles: apoptotic bodies, microvesicles, and exosomes. Prostate 74(14):1379–1390. doi: 10.1002/pros.22853 CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    van der Goot FG, Gruenberg J (2006) Intra-endosomal membrane traffic. Trends Cell Biol 16(10):514–521. doi: 10.1016/j.tcb.2006.08.003 CrossRefPubMedGoogle Scholar
  16. 16.
    Tamai K, Tanaka N, Nakano T, Kakazu E, Kondo Y, Inoue J, Shiina M, Fukushima K, Hoshino T, Sano K, Ueno Y, Shimosegawa T, Sugamura K (2010) Exosome secretion of dendritic cells is regulated by Hrs, an ESCRT-0 protein. Biochem Biophys Res Commun 399(3):384–390. doi: 10.1016/j.bbrc.2010.07.083 CrossRefPubMedGoogle Scholar
  17. 17.
    Colombo M, Moita C, van Niel G, Kowal J, Vigneron J, Benaroch P, Manel N, Moita LF, Thery C, Raposo G (2013) Analysis of ESCRT functions in exosome biogenesis, composition and secretion highlights the heterogeneity of extracellular vesicles. J Cell Sci 126(Pt 24):5553–5565. doi: 10.1242/jcs.128868 CrossRefPubMedGoogle Scholar
  18. 18.
    Baietti MF, Zhang Z, Mortier E, Melchior A, Degeest G, Geeraerts A, Ivarsson Y, Depoortere F, Coomans C, Vermeiren E, Zimmermann P, David G (2012) Syndecan-syntenin-ALIX regulates the biogenesis of exosomes. Nat Cell Biol 14(7):677–685. doi: 10.1038/ncb2502 CrossRefPubMedGoogle Scholar
  19. 19.
    Chairoungdua A, Smith DL, Pochard P, Hull M, Caplan MJ (2010) Exosome release of beta-catenin: a novel mechanism that antagonizes Wnt signaling. J Cell Biol 190(6):1079–1091. doi: 10.1083/jcb.201002049 CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Nazarenko I, Rana S, Baumann A, McAlear J, Hellwig A, Trendelenburg M, Lochnit G, Preissner KT, Zoller M (2010) Cell surface tetraspanin Tspan8 contributes to molecular pathways of exosome-induced endothelial cell activation. Cancer Res 70(4):1668–1678. doi: 10.1158/0008-5472.can-09-2470 CrossRefPubMedGoogle Scholar
  21. 21.
    Trajkovic K, Hsu C, Chiantia S, Rajendran L, Wenzel D, Wieland F, Schwille P, Brugger B, Simons M (2008) Ceramide triggers budding of exosome vesicles into multivesicular endosomes. Science 319(5867):1244–1247. doi: 10.1126/science.1153124 CrossRefPubMedGoogle Scholar
  22. 22.
    Phuyal S, Hessvik NP, Skotland T, Sandvig K, Llorente A (2014) Regulation of exosome release by glycosphingolipids and flotillins. FEBS J 281(9):2214–2227. doi: 10.1111/febs.12775 CrossRefPubMedGoogle Scholar
  23. 23.
    Ostrowski M, Carmo NB, Krumeich S, Fanget I, Raposo G, Savina A, Moita CF, Schauer K, Hume AN, Freitas RP, Goud B, Benaroch P, Hacohen N, Fukuda M, Desnos C, Seabra MC, Darchen F, Amigorena S, Moita LF, Thery C (2010) Rab27a and Rab27b control different steps of the exosome secretion pathway. Nat Cell Biol 12(1):19–30. doi: 10.1038/ncb2000 (sup pp 11–13) CrossRefPubMedGoogle Scholar
  24. 24.
    Savina A, Fader CM, Damiani MT, Colombo MI (2005) Rab11 promotes docking and fusion of multivesicular bodies in a calcium-dependent manner. Traffic 6(2):131–143. doi: 10.1111/j.1600-0854.2004.00257.x CrossRefPubMedGoogle Scholar
  25. 25.
    Hsu C, Morohashi Y, Yoshimura S, Manrique-Hoyos N, Jung S, Lauterbach MA, Bakhti M, Gronborg M, Mobius W, Rhee J, Barr FA, Simons M (2010) Regulation of exosome secretion by Rab35 and its GTPase-activating proteins TBC1D10A-C. J Cell Biol 189(2):223–232. doi: 10.1083/jcb.200911018 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Baixauli F, Lopez-Otin C, Mittelbrunn M (2014) Exosomes and autophagy: coordinated mechanisms for the maintenance of cellular fitness. Front Immunol 5:403. doi: 10.3389/fimmu.2014.00403 CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Boya P, Reggiori F, Codogno P (2013) Emerging regulation and functions of autophagy. Nat Cell Biol 15(7):713–720. doi: 10.1038/ncb2788 CrossRefPubMedGoogle Scholar
  28. 28.
    Ponpuak M, Mandell MA, Kimura T, Chauhan S, Cleyrat C, Deretic V (2015) Secretory autophagy. Curr Opin Cell Biol 35:106–116. doi: 10.1016/ CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Ejlerskov P, Rasmussen I, Nielsen TT, Bergstrom AL, Tohyama Y, Jensen PH, Vilhardt F (2013) Tubulin polymerization-promoting protein (TPPP/p25alpha) promotes unconventional secretion of alpha-synuclein through exophagy by impairing autophagosome-lysosome fusion. J Biol Chem 288(24):17313–17335. doi: 10.1074/jbc.M112.401174 CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Nilsson P, Loganathan K, Sekiguchi M, Matsuba Y, Hui K, Tsubuki S, Tanaka M, Iwata N, Saito T, Saido TC (2013) Abeta secretion and plaque formation depend on autophagy. Cell reports 5(1):61–69. doi: 10.1016/j.celrep.2013.08.042 CrossRefPubMedGoogle Scholar
  31. 31.
    Di Paolo G, De Camilli P (2006) Phosphoinositides in cell regulation and membrane dynamics. Nature 443(7112):651–657. doi: 10.1038/nature05185 CrossRefPubMedGoogle Scholar
  32. 32.
    Burman C, Ktistakis NT (2010) Regulation of autophagy by phosphatidylinositol 3-phosphate. FEBS Lett 584(7):1302–1312. doi: 10.1016/j.febslet.2010.01.011 CrossRefPubMedGoogle Scholar
  33. 33.
    Dove SK, Cooke FT, Douglas MR, Sayers LG, Parker PJ, Michell RH (1997) Osmotic stress activates phosphatidylinositol-3,5-bisphosphate synthesis. Nature 390(6656):187–192. doi: 10.1038/36613 CrossRefPubMedGoogle Scholar
  34. 34.
    Whiteford CC, Brearley CA, Ulug ET (1997) Phosphatidylinositol 3,5-bisphosphate defines a novel PI 3-kinase pathway in resting mouse fibroblasts. Biochem J 323(Pt 3):597–601CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Li X, Wang X, Zhang X, Zhao M, Tsang WL, Zhang Y, Yau RG, Weisman LS, Xu H (2013) Genetically encoded fluorescent probe to visualize intracellular phosphatidylinositol 3,5-bisphosphate localization and dynamics. Proc Natl Acad Sci USA 110(52):21165–21170. doi: 10.1073/pnas.1311864110 CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Shisheva A, Sbrissa D, Ikonomov O (2015) Plentiful PtdIns5P from scanty PtdIns(3,5)P or from ample PtdIns? PIKfyve-dependent models: evidence and speculation (response to: DOI 10.1002/bies.201300012). Bioessays 37(3):267–277. doi: 10.1002/bies.201400129 CrossRefPubMedGoogle Scholar
  37. 37.
    McCartney AJ, Zhang Y, Weisman LS (2014) Phosphatidylinositol 3,5-bisphosphate: low abundance, high significance. BioEssays 36(1):52–64. doi: 10.1002/bies.201300012 CrossRefPubMedGoogle Scholar
  38. 38.
    Yamamoto A, DeWald DB, Boronenkov IV, Anderson RA, Emr SD, Koshland D (1995) Novel PI(4)P 5-kinase homologue, Fab1p, essential for normal vacuole function and morphology in yeast. Mol Biol Cell 6(5):525–539CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Rutherford AC, Traer C, Wassmer T, Pattni K, Bujny MV, Carlton JG, Stenmark H, Cullen PJ (2006) The mammalian phosphatidylinositol 3-phosphate 5-kinase (PIKfyve) regulates endosome-to-TGN retrograde transport. J Cell Sci 119(Pt 19):3944–3957. doi: 10.1242/jcs.03153 CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Jefferies HB, Cooke FT, Jat P, Boucheron C, Koizumi T, Hayakawa M, Kaizawa H, Ohishi T, Workman P, Waterfield MD, Parker PJ (2008) A selective PIKfyve inhibitor blocks PtdIns(3,5)P(2) production and disrupts endomembrane transport and retroviral budding. EMBO Rep 9(2):164–170. doi: 10.1038/sj.embor.7401155 CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Cai X, Xu Y, Cheung AK, Tomlinson RC, Alcazar-Roman A, Murphy L, Billich A, Zhang B, Feng Y, Klumpp M, Rondeau JM, Fazal AN, Wilson CJ, Myer V, Joberty G, Bouwmeester T, Labow MA, Finan PM, Porter JA, Ploegh HL, Baird D, De Camilli P, Tallarico JA, Huang Q (2013) PIKfyve, a class III PI kinase, is the target of the small molecular IL-12/IL-23 inhibitor apilimod and a player in Toll-like receptor signaling. Chem Biol 20(7):912–921. doi: 10.1016/j.chembiol.2013.05.010 CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Martin S, Harper CB, May LM, Coulson EJ, Meunier FA, Osborne SL (2013) Inhibition of PIKfyve by YM-201636 dysregulates autophagy and leads to apoptosis-independent neuronal cell death. PLoS One 8(3):e60152. doi: 10.1371/journal.pone.0060152 CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Rusten TE, Vaccari T, Lindmo K, Rodahl LM, Nezis IP, Sem-Jacobsen C, Wendler F, Vincent JP, Brech A, Bilder D, Stenmark H (2007) ESCRTs and Fab1 regulate distinct steps of autophagy. Current Biol: CB 17(20):1817–1825. doi: 10.1016/j.cub.2007.09.032 CrossRefGoogle Scholar
  44. 44.
    Vicinanza M, Korolchuk VI, Ashkenazi A, Puri C, Menzies FM, Clarke JH, Rubinsztein DC (2015) PI(5)P regulates autophagosome biogenesis. Mol Cell 57(2):219–234. doi: 10.1016/j.molcel.2014.12.007 CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    de Lartigue J, Polson H, Feldman M, Shokat K, Tooze SA, Urbe S, Clague MJ (2009) PIKfyve regulation of endosome-linked pathways. Traffic 10(7):883–893CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Osborne SL, Wen PJ, Boucheron C, Nguyen HN, Hayakawa M, Kaizawa H, Parker PJ, Vitale N, Meunier FA (2008) PIKfyve negatively regulates exocytosis in neurosecretory cells. J Biol Chem 283(5):2804–2813. doi: 10.1074/jbc.M704856200 CrossRefPubMedGoogle Scholar
  47. 47.
    Messenger SW, Thomas DD, Cooley MM, Jones EK, Falkowski MA, August BK, Fernandez LA, Gorelick FS, Groblewski GE (2015) Early to late endosome trafficking controls secretion and zymogen activation in rodent and human pancreatic acinar cells. Cell Mol Gastroenterol Hepatol 1(6):695–709. doi: 10.1016/j.jcmgh.2015.08.002 CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Llorente A, de Marco MC, Alonso MA (2004) Caveolin-1 and MAL are located on prostasomes secreted by the prostate cancer PC-3 cell line. J Cell Sci 117(22):5343–5351. doi: 10.1242/jcs.01420 CrossRefPubMedGoogle Scholar
  49. 49.
    Llorente A, van Deurs B, Sandvig K (2007) Cholesterol regulates prostasome release from secretory lysosomes in PC-3 human prostate cancer cells. Eur J Cell Biol 86(7):405–415CrossRefPubMedGoogle Scholar
  50. 50.
    Phuyal S, Skotland T, Hessvik NP, Simolin H, Overbye A, Brech A, Parton RG, Ekroos K, Sandvig K, Llorente A (2015) The ether lipid precursor hexadecylglycerol stimulates the release and changes the composition of exosomes derived from PC-3 cells. J Biol Chem 13(290):4225–4237. doi: 10.1074/jbc.M114.593962 CrossRefGoogle Scholar
  51. 51.
    Slot JW, Geuze HJ, Gigengack S, Lienhard GE, James DE (1991) Immuno-localization of the insulin regulatable glucose transporter in brown adipose tissue of the rat. J Cell Biol 113(1):123–135CrossRefPubMedGoogle Scholar
  52. 52.
    Filimonenko M, Stuffers S, Raiborg C, Yamamoto A, Malerod L, Fisher EM, Isaacs A, Brech A, Stenmark H, Simonsen A (2007) Functional multivesicular bodies are required for autophagic clearance of protein aggregates associated with neurodegenerative disease. J Cell Biol 179(3):485–500. doi: 10.1083/jcb.200702115 CrossRefPubMedPubMedCentralGoogle Scholar
  53. 53.
    Keller A, Nesvizhskii AI, Kolker E, Aebersold R (2002) Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem 74(20):5383–5392CrossRefPubMedGoogle Scholar
  54. 54.
    Nesvizhskii AI, Keller A, Kolker E, Aebersold R (2003) A statistical model for identifying proteins by tandem mass spectrometry. Anal Chem 75(17):4646–4658CrossRefPubMedGoogle Scholar
  55. 55.
    Johansen T, Lamark T (2011) Selective autophagy mediated by autophagic adapter proteins. Autophagy 7(3):279–296CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Kabeya Y, Mizushima N, Yamamoto A, Oshitani-Okamoto S, Ohsumi Y, Yoshimori T (2004) LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formation. J Cell Sci 117(Pt 13):2805–2812. doi: 10.1242/jcs.01131 CrossRefPubMedGoogle Scholar
  57. 57.
    Polson HE, de Lartigue J, Rigden DJ, Reedijk M, Urbe S, Clague MJ, Tooze SA (2010) Mammalian Atg18 (WIPI2) localizes to omegasome-anchored phagophores and positively regulates LC3 lipidation. Autophagy 6(4):506–522. doi: 10.4161/auto.6.4.11863 CrossRefPubMedGoogle Scholar
  58. 58.
    Dooley HC, Razi M, Polson HE, Girardin SE, Wilson MI, Tooze SA (2014) WIPI2 links LC3 conjugation with PI3P, autophagosome formation, and pathogen clearance by recruiting Atg12-5-16L1. Mol Cell 55(2):238–252. doi: 10.1016/j.molcel.2014.05.021 CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Nanao T, Koike M, Yamaguchi J, Sasaki M, Uchiyama Y (2015) Cellular localization and tissue distribution of endogenous DFCP1 protein. Biomed Res (Tokyo, Japan) 36(2):121–133CrossRefGoogle Scholar
  60. 60.
    Kharaziha P, Chioureas D, Rutishauser D, Baltatzis G, Lennartsson L, Fonseca P, Azimi A, Hultenby K, Zubarev R, Ullen A, Yachnin J, Nilsson S, Panaretakis T (2015) Molecular profiling of prostate cancer derived exosomes may reveal a predictive signature for response to docetaxel. Oncotarget 6(25):21740–21754. doi: 10.18632/oncotarget.3226 CrossRefPubMedPubMedCentralGoogle Scholar
  61. 61.
    Welton JL, Khanna S, Giles PJ, Brennan P, Brewis IA, Staffurth J, Mason MD, Clayton A (2010) Proteomics analysis of bladder cancer exosomes. Mol Cell Proteom: MCP 9(6):1324–1338. doi: 10.1074/mcp.M000063-MCP201 CrossRefGoogle Scholar
  62. 62.
    Cruceanu C, Alda M, Turecki G (2009) Lithium: a key to the genetics of bipolar disorder. Genome medicine 1(8):79. doi: 10.1186/gm79 CrossRefPubMedPubMedCentralGoogle Scholar
  63. 63.
    Xu J, Wong C (2008) A computational screen for mouse signaling pathways targeted by microRNA clusters. RNA (New York, NY) 14(7):1276–1283. doi: 10.1261/rna.997708 CrossRefGoogle Scholar
  64. 64.
    Thery C, Zitvogel L, Amigorena S (2002) Exosomes: composition, biogenesis and function. Nat Rev Immunol 2(8):569–579PubMedGoogle Scholar
  65. 65.
    Dupont N, Jiang S, Pilli M, Ornatowski W, Bhattacharya D, Deretic V (2011) Autophagy-based unconventional secretory pathway for extracellular delivery of IL-1beta. EMBO J 30(23):4701–4711. doi: 10.1038/emboj.2011.398 CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Duran JM, Anjard C, Stefan C, Loomis WF, Malhotra V (2010) Unconventional secretion of Acb1 is mediated by autophagosomes. J Cell Biol 188(4):527–536. doi: 10.1083/jcb.200911154 CrossRefPubMedPubMedCentralGoogle Scholar
  67. 67.
    Gerstenmaier L, Pilla R, Herrmann L, Herrmann H, Prado M, Villafano GJ, Kolonko M, Reimer R, Soldati T, King JS, Hagedorn M (2015) The autophagic machinery ensures nonlytic transmission of mycobacteria. Proc Natl Acad Sci USA 112(7):E687–E692. doi: 10.1073/pnas.1423318112 CrossRefPubMedPubMedCentralGoogle Scholar
  68. 68.
    Buschow SI, Liefhebber JM, Wubbolts R, Stoorvogel W (2005) Exosomes contain ubiquitinated proteins. Blood Cells Mol Dis 35(3):398–403. doi: 10.1016/j.bcmd.2005.08.005 CrossRefPubMedGoogle Scholar
  69. 69.
    Ikonomov OC, Sbrissa D, Shisheva A (2006) Localized PtdIns 3,5-P2 synthesis to regulate early endosome dynamics and fusion. Am J Physiol Cell Physiol 291(2):C393–C404. doi: 10.1152/ajpcell.00019.2006 CrossRefPubMedGoogle Scholar
  70. 70.
    Fader CM, Sanchez D, Furlan M, Colombo MI (2008) Induction of autophagy promotes fusion of multivesicular bodies with autophagic vacuoles in k562 cells. Traffic 9(2):230–250. doi: 10.1111/j.1600-0854.2007.00677.x CrossRefPubMedGoogle Scholar
  71. 71.
    Sahu R, Kaushik S, Clement CC, Cannizzo ES, Scharf B, Follenzi A, Potolicchio I, Nieves E, Cuervo AM, Santambrogio L (2011) Microautophagy of cytosolic proteins by late endosomes. Dev Cell 20(1):131–139. doi: 10.1016/j.devcel.2010.12.003 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer International Publishing 2016

Authors and Affiliations

  • Nina Pettersen Hessvik
    • 1
    • 2
  • Anders Øverbye
    • 1
    • 2
  • Andreas Brech
    • 1
    • 2
    • 3
  • Maria Lyngaas Torgersen
    • 1
    • 2
  • Ida Seim Jakobsen
    • 1
    • 2
  • Kirsten Sandvig
    • 1
    • 2
    • 3
  • Alicia Llorente
    • 1
    • 2
    Email author
  1. 1.Department of Molecular Cell Biology, The Norwegian Radium Hospital, Institute for Cancer ResearchOslo University HospitalOsloNorway
  2. 2.Centre for Cancer BiomedicineUniversity of OsloOsloNorway
  3. 3.Department of BiosciencesUniversity of OsloOsloNorway

Personalised recommendations