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Cited by 447Extracellular vesicles in diseaseCellular transport and secretionNanoparticle-Based Drug Delivery

Exosomes surf on filopodia to enter cells at endocytic hot spots, traffic within endosomes, and are targeted to the ER

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Heusermann, Wolf, Hean, Justin, Trojer, Dominic, Steib, Emmanuelle, von Bueren, Stefan, Graff-Meyer, Alexandra, Genoud, Christel, Martin, Katrin +6 more·2016·Journal of Cell Biology
DOI 10.1083/jcb.201506084PMC PMC5084269
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Abstract

Exosomes are nanovesicles released by virtually all cells, which act as intercellular messengers by transfer of protein, lipid, and RNA cargo. Their quantitative efficiency, routes of cell uptake, and subcellular fate within recipient cells remain elusive. We quantitatively characterize exosome cell uptake, which saturates with dose and time and reaches near 100% transduction efficiency at picomolar concentrations. Highly reminiscent of pathogenic bacteria and viruses, exosomes are recruited as single vesicles to the cell body by surfing on filopodia as well as filopodia grabbing and pulling motions to reach endocytic hot spots at the filopodial base. After internalization, exosomes shuttle within endocytic vesicles to scan the endoplasmic reticulum before being sorted into the lysosome as their final intracellular destination. Our data quantify and explain the efficiency of exosome internalization by recipient cells, establish a new parallel between exosome and virus host cell interaction, and suggest unanticipated routes of subcellular cargo delivery.

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Crossref · 10.1083/jcb.201506084

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  • Endocytosed EVs in large endosomal compartments traffic towards the endoplasmic reticulum and finally sort into late-endosomes.

    “Recently a detailed study has demonstrated that endocytosed EVs in large endosomal compartments traffic towards the endoplasmic reticulum and finally sort into late-endosomes67.”

    From GAPDH controls extracellular vesicle biogenesis and enhances the therapeutic potential of EV mediated siRNA delivery to the brain (2021) · cites this paper as [67]

    The Results and discussion section reports that internalized exosomes travel within larger endocytic vesicles and move along the ER. It describes their subsequent sorting more specifically as movement to lysosomes, with increasing lysosomal colocalization over time but minimal colocalization with the multivesicular-body marker. Thus the claim's final destination, stated simply as late endosomes, is imprecise. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 46

  1. Alvarez-Erviti L., Seow Y., Yin H., Betts C., Lakhal S., and Wood M.J.A. 2011 Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat. Biotechnol. 29: 341– 345. 10.1038/nbt.1807 21423189 · DOI · Paper page
  2. Barry D.J., Durkin C.H., Abella J.V., and Way M. 2015 Open source software for quantification of cell migration, protrusions, and fluorescence intensities. J. Cell Biol. 209: 163– 180. 10.1083/jcb.201501081 25847537 PMC4395480 · DOI
  3. Bolte S., and Cordelières F.P. 2006 A guided tour into subcellular colocalization analysis in light microscopy. J. Microsc. 224: 213– 232. 10.1111/j.1365-2818.2006.01706.x 17210054 · DOI
  4. Bornschlögl T. 2013 How filopodia pull: what we know about the mechanics and dynamics of filopodia. Cytoskeleton (Hoboken). 70: 590– 603. 10.1002/cm.21130 23959922 · DOI
  5. Chevillet J.R., Kang Q., Ruf I.K., Briggs H.A., Vojtech L.N., Hughes S.M., Cheng H.H., Arroyo J.D., Meredith E.K., Gallichotte E.N., 2014 Quantitative and stoichiometric analysis of the microRNA content of exosomes. Proc. Natl. Acad. Sci. USA. 111: 14888– 14893. 10.1073/pnas.1408301111 25267620 PMC4205618 · DOI
  6. Christianson H.C., Svensson K.J., van Kuppevelt T.H., Li J.-P., and Belting M. 2013 Cancer cell exosomes depend on cell-surface heparan sulfate proteoglycans for their internalization and functional activity. Proc. Natl. Acad. Sci. USA. 110: 17380– 17385. 10.1073/pnas.1304266110 24101524 PMC3808637 · DOI · Paper page
  7. Colombo M., Raposo G., and Théry C. 2014 Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu. Rev. Cell Dev. Biol. 30: 255– 289. 10.1146/annurev-cellbio-101512-122326 25288114 · DOI · Paper page
  8. del Conde I., Shrimpton C.N., Thiagarajan P., and López J.A. 2005 Tissue-factor-bearing microvesicles arise from lipid rafts and fuse with activated platelets to initiate coagulation. Blood. 106: 1604– 1611. 10.1182/blood-2004-03-1095 15741221 · DOI
  9. Feng D., Zhao W.-L., Ye Y.-Y., Bai X.-C., Liu R.-Q., Chang L.-F., Zhou Q., and Sui S.-F. 2010 Cellular internalization of exosomes occurs through phagocytosis. Traffic. 11: 675– 687. 10.1111/j.1600-0854.2010.01041.x 20136776 · DOI
  10. Fitzner D., Schnaars M., van Rossum D., Krishnamoorthy G., Dibaj P., Bakhti M., Regen T., Hanisch U.-K., and Simons M. 2011 Selective transfer of exosomes from oligodendrocytes to microglia by macropinocytosis. J. Cell Sci. 124: 447– 458. 10.1242/jcs.074088 21242314 · DOI · Paper page
  11. Forscher P., and Smith S.J. 1988 Actions of cytochalasins on the organization of actin filaments and microtubules in a neuronal growth cone. J. Cell Biol. 107: 1505– 1516. 10.1083/jcb.107.4.1505 3170637 PMC2115246 · DOI · Paper page
  12. Friedman J.R., Dibenedetto J.R., West M., Rowland A.A., and Voeltz G.K. 2013 Endoplasmic reticulum-endosome contact increases as endosomes traffic and mature. Mol. Biol. Cell. 24: 1030– 1040. 10.1091/mbc.E12-10-0733 23389631 PMC3608491 · DOI
  13. Gilleron J., Querbes W., Zeigerer A., Borodovsky A., Marsico G., Schubert U., Manygoats K., Seifert S., Andree C., Stöter M., 2013 Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape. Nat. Biotechnol. 31: 638– 646. 10.1038/nbt.2612 23792630 · DOI · Paper page
  14. Hood J.L., San R.S., and Wickline S.A. 2011 Exosomes released by melanoma cells prepare sentinel lymph nodes for tumor metastasis. Cancer Res. 71: 3792– 3801. 10.1158/0008-5472.CAN-10-4455 21478294 · DOI
  15. Isogai T., van der Kammen R., and Innocenti M. 2015 SMIFH2 has effects on Formins and p53 that perturb the cell cytoskeleton. Sci. Rep. 5: 9802 10.1038/srep09802 25925024 PMC5386218 · DOI
  16. Jagannathan S., Reid D.W., Cox A.H., and Nicchitta C.V. 2014 De novo translation initiation on membrane-bound ribosomes as a mechanism for localization of cytosolic protein mRNAs to the endoplasmic reticulum. RNA. 20: 1489– 1498. 10.1261/rna.045526.114 25142066 PMC4174431 · DOI
  17. Kordelas L., Rebmann V., Ludwig A.-K., Radtke S., Ruesing J., Doeppner T.R., Epple M., Horn P.A., Beelen D.W., and Giebel B. 2014 MSC-derived exosomes: a novel tool to treat therapy-refractory graft-versus-host disease. Leukemia. 28: 970– 973. 24445866
  18. Lam S.S., Martell J.D., Kamer K.J., Deerinck T.J., Ellisman M.H., Mootha V.K., and Ting A.Y. 2015 Directed evolution of APEX2 for electron microscopy and proximity labeling. Nat. Methods. 12: 51– 54. 10.1038/nmeth.3179 25419960 PMC4296904 · DOI
  19. Lehmann M.J., Sherer N.M., Marks C.B., Pypaert M., and Mothes W. 2005 Actin- and myosin-driven movement of viruses along filopodia precedes their entry into cells. J. Cell Biol. 170: 317– 325. 10.1083/jcb.200503059 16027225 PMC2171413 · DOI · Paper page
  20. Li S., Liu L., Zhuang X., Yu Y., Liu X., Cui X., Ji L., Pan Z., Cao X., Mo B., 2013 MicroRNAs inhibit the translation of target mRNAs on the endoplasmic reticulum in Arabidopsis. Cell. 153: 562– 574. 10.1016/j.cell.2013.04.005 23622241 PMC3694718 · DOI
  21. Lidke D.S., Lidke K.A., Rieger B., Jovin T.M., and Arndt-Jovin D.J. 2005 Reaching out for signals: filopodia sense EGF and respond by directed retrograde transport of activated receptors. J. Cell Biol. 170: 619– 626. 10.1083/jcb.200503140 16103229 PMC2171515 · DOI
  22. Martin K., Vilela M., Jeon N.L., Danuser G., and Pertz O. 2014 A growth factor-induced, spatially organizing cytoskeletal module enables rapid and persistent fibroblast migration. Dev. Cell. 30: 701– 716. 10.1016/j.devcel.2014.07.022 25268172 PMC4385272 · DOI
  23. Mattila P.K., and Lappalainen P. 2008 Filopodia: molecular architecture and cellular functions. Nat. Rev. Mol. Cell Biol. 9: 446– 454. 10.1038/nrm2406 18464790 · DOI
  24. Mitchison T., and Kirschner M. 1988 Cytoskeletal dynamics and nerve growth. Neuron. 1: 761– 772. 10.1016/0896-6273(88)90124-9 3078414 · DOI
  25. Morelli A.E., Larregina A.T., Shufesky W.J., Sullivan M.L.G., Stolz D.B., Papworth G.D., Zahorchak A.F., Logar A.J., Wang Z., Watkins S.C., 2004 Endocytosis, intracellular sorting, and processing of exosomes by dendritic cells. Blood. 104: 3257– 3266. 10.1182/blood-2004-03-0824 15284116 · DOI
  26. Nobile C., Rudnicka D., Hasan M., Aulner N., Porrot F., Machu C., Renaud O., Prévost M.-C., Hivroz C., Schwartz O., and Sol-Foulon N. 2010 HIV-1 Nef inhibits ruffles, induces filopodia, and modulates migration of infected lymphocytes. J. Virol. 84: 2282– 2293. 10.1128/JVI.02230-09 20015995 PMC2820911 · DOI
  27. Nordin J.Z., Lee Y., Vader P., Mäger I., Johansson H.J., Heusermann W., Wiklander O.P.B., Hällbrink M., Seow Y., Bultema J.J., 2015 Ultrafiltration with size-exclusion liquid chromatography for high yield isolation of extracellular vesicles preserving intact biophysical and functional properties. Nanomedicine (Lond.). 11: 879– 883. 25659648
  28. Parolini I., Federici C., Raggi C., Lugini L., Palleschi S., De Milito A., Coscia C., Iessi E., Logozzi M., Molinari A., 2009 Microenvironmental pH is a key factor for exosome traffic in tumor cells. J. Biol. Chem. 284: 34211– 34222. 10.1074/jbc.M109.041152 19801663 PMC2797191 · DOI
  29. Properzi F., Logozzi M., and Fais S. 2013 Exosomes: the future of biomarkers in medicine. Biomarkers Med. 7: 769– 778. 10.2217/bmm.13.63 24044569 · DOI
  30. Rana S., and Zöller M. 2011 Exosome target cell selection and the importance of exosomal tetraspanins: a hypothesis. Biochem. Soc. Trans. 39: 559– 562. 10.1042/BST0390559 21428939 · DOI
  31. Ratajczak J., Miekus K., Kucia M., Zhang J., Reca R., Dvorak P., and Ratajczak M.Z. 2006 Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: evidence for horizontal transfer of mRNA and protein delivery. Leukemia. 20: 847– 856. 10.1038/sj.leu.2404132 16453000 · DOI
  32. Reid D.W., and Nicchitta C.V. 2015 Diversity and selectivity in mRNA translation on the endoplasmic reticulum. Nat. Rev. Mol. Cell Biol. 16: 221– 231. 25735911 PMC4494666
  33. Reid D.W., Chen Q., Tay A.S.-L., Shenolikar S., and Nicchitta C.V. 2014 The unfolded protein response triggers selective mRNA release from the endoplasmic reticulum. Cell. 158: 1362– 1374. 10.1016/j.cell.2014.08.012 25215492 PMC4163055 · DOI
  34. Rizvi S.A., Neidt E.M., Cui J., Feiger Z., Skau C.T., Gardel M.L., Kozmin S.A., and Kovar D.R. 2009 Identification and characterization of a small molecule inhibitor of formin-mediated actin assembly. Chem. Biol. 16: 1158– 1168. 10.1016/j.chembiol.2009.10.006 19942139 PMC2784894 · DOI
  35. Sahay G., Querbes W., Alabi C., Eltoukhy A., Sarkar S., Zurenko C., Karagiannis E., Love K., Chen D., Zoncu R., 2013 Efficiency of siRNA delivery by lipid nanoparticles is limited by endocytic recycling. Nat. Biotechnol. 31: 653– 658. 10.1038/nbt.2614 23792629 PMC3814166 · DOI · Paper page
  36. Sheetz M.P., Turney S., Qian H., and Elson E.L. 1989 Nanometre-level analysis demonstrates that lipid flow does not drive membrane glycoprotein movements. Nature. 340: 284– 288. 10.1038/340284a0 2747796 · DOI
  37. Smith J.L., Campos S.K., Wandinger-Ness A., and Ozbun M.A. 2008 Caveolin-1-dependent infectious entry of human papillomavirus type 31 in human keratinocytes proceeds to the endosomal pathway for pH-dependent uncoating. J. Virol. 82: 9505– 9512. 10.1128/JVI.01014-08 18667513 PMC2546945 · DOI
  38. Stalder L., Heusermann W., Sokol L., Trojer D., Wirz J., Hean J., Fritzsche A., Aeschimann F., Pfanzagl V., Basselet P., 2013 The rough endoplasmatic reticulum is a central nucleation site of siRNA-mediated RNA silencing. EMBO J. 32: 1115– 1127. 10.1038/emboj.2013.52 23511973 PMC3630355 · DOI
  39. Svensson K.J., Christianson H.C., Wittrup A., Bourseau-Guilmain E., Lindqvist E., Svensson L.M., Mörgelin M., and Belting M. 2013 Exosome uptake depends on ERK1/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1. J. Biol. Chem. 288: 17713– 17724. 10.1074/jbc.M112.445403 23653359 PMC3682571 · DOI
  40. Thayanithy V., Babatunde V., Dickson E.L., Wong P., Oh S., Ke X., Barlas A., Fujisawa S., Romin Y., Moreira A.L., 2014 Tumor exosomes induce tunneling nanotubes in lipid raft-enriched regions of human mesothelioma cells. Exp. Cell Res. 323: 178– 188. 10.1016/j.yexcr.2014.01.014 24468420 PMC4159162 · DOI
  41. Tian T., Zhu Y.-L., Hu F.-H., Wang Y.-Y., Huang N.-P., and Xiao Z.-D. 2013 Dynamics of exosome internalization and trafficking. J. Cell. Physiol. 228: 1487– 1495. 10.1002/jcp.24304 23254476 · DOI
  42. Tian T., Zhu Y.-L., Zhou Y.-Y., Liang G.-F., Wang Y.-Y., Hu F.-H., and Xiao Z.-D. 2014 Exosome uptake through clathrin-mediated endocytosis and macropinocytosis and mediating miR-21 delivery. J. Biol. Chem. 289: 22258– 22267. 10.1074/jbc.M114.588046 24951588 PMC4139237 · DOI · Paper page
  43. Twu O., de Miguel N., Lustig G., Stevens G.C., Vashisht A.A., Wohlschlegel J.A., and Johnson P.J. 2013 Trichomonas vaginalis exosomes deliver cargo to host cells and mediate host:parasite interactions. PLoS Pathog. 9: e1003482 10.1371/journal.ppat.1003482 23853596 PMC3708881 · DOI
  44. Valadi H., Ekström K., Bossios A., Sjöstrand M., Lee J.J., and Lötvall J.O. 2007 Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat. Cell Biol. 9: 654– 659. 10.1038/ncb1596 17486113 · DOI · Paper page
  45. Wurdinger T., Gatson N.N., Balaj L., Kaur B., Breakefield X.O., and Pegtel D.M. 2012 Extracellular vesicles and their convergence with viral pathways. Adv. Virol. 2012: 767694 10.1155/2012/767694 22888349 PMC3410301 · DOI
  46. Zamudio-Meza H., Castillo-Alvarez A., González-Bonilla C., and Meza I. 2009 Cross-talk between Rac1 and Cdc42 GTPases regulates formation of filopodia required for dengue virus type-2 entry into HMEC-1 cells. J. Gen. Virol. 90: 2902– 2911. 10.1099/vir.0.014159-0 19710257 · DOI

Cited by

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Authors
Heusermann, W., Hean, J., Trojer, D., Steib, E., von Bueren, S., Graff-Meyer, A., Genoud, C., Martin, K., Pizzato, N., Voshol, J., Morrissey, D. V., Andaloussi, S. E. L., Wood, M. J., & Meisner-Kober, N. C.
Affiliations
  • Novartis (Switzerland) (CH)
  • Novartis Institutes for Biomedical Research, CH-4000 Basel, Switzerland 1
  • Friedrich Miescher Institute (CH)
  • University of Basel (CH)
  • Novartis Institutes for Biomedical Research, Cambridge, MA 02139 5
  • Karolinska Institutet (SE)
  • University of Oxford (GB)
Corresponding author
Heusermann, Wolf
ORCID
Venue
Journal of Cell Biology
Publisher
Rockefeller University Press
Published
2016-04-25
Volume
213
Issue
2
Pages
173-184
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
DOI
10.1083/jcb.201506084
PMC
PMC5084269
OpenAlex
W2342982379
Cited by
447
References
1
Citations by year
2026: 27 · 2025: 40 · 2024: 40 · 2023: 51 · 2022: 49 · 2021: 52 · 2020: 64 · 2019: 42 · 2018: 32 · 2017: 38 · 2016: 12
FWCI
21.53
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • Spiral (Imperial College London) (submittedVersion, cc-by-nc-sa)
Funders
  • Vetenskapsrådet (VR-Med, EuroNanoMedII)
  • Swedish Society for Medical Research
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
exosomes, exosome uptake, filopodia, endocytosis, endosomes, endoplasmic reticulum targeting
MeSH terms
Biological Transport, Endoplasmic Reticulum, Humans, Microscopy, Electron, Scanning, Pseudopodia, Endosomes, Exosomes, HEK293 Cells

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