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The power of imaging to understand extracellular vesicle biology in vivo

Nature Methods volume 18, pages 1013–1026 (2021)Cite this article

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Abstract

Extracellular vesicles (EVs) are nano-sized lipid bilayer vesicles released by virtually every cell type. EVs have diverse biological activities, ranging from roles in development and homeostasis to cancer progression, which has spurred the development of EVs as disease biomarkers and drug nanovehicles. Owing to the small size of EVs, however, most studies have relied on isolation and biochemical analysis of bulk EVs separated from biofluids. Although informative, these approaches do not capture the dynamics of EV release, biodistribution, and other contributions to pathophysiology. Recent advances in live and high-resolution microscopy techniques, combined with innovative EV labeling strategies and reporter systems, provide new tools to study EVs in vivo in their physiological environment and at the single-vesicle level. Here we critically review the latest advances and challenges in EV imaging, and identify urgent, outstanding questions in our quest to unravel EV biology and therapeutic applications.

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Fig. 1: Timeline of EV imaging milestones and broad overview of microscopy techniques to resolve EVs at different scales.
Fig. 2: Tagging strategies to image EV production.
Fig. 3: Imaging EV propagation in vivo.
Fig. 4: Tagging strategies to image EV interaction, uptake, and fate.

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Article 01 December 2022

Zixuan Zhao, Xinyi Chen, … Hanry Yu

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Acknowledgements

The authors acknowledge financial support from the International Society for Extracellular Vesicles, the French Society of Extracellular vesicles, the Société Française des Microscopies, the ITMO BCDE for their support for the organization of the ISEV workshop ‘EV imaging in vivo’ that provided the basis for this review. We thank the workshop organizing committee members G.D.’A., V.H., E.-M.K.-A., X.L., and K.W. for their help. We thank P. Stahl (Washington University School of Medicine, USA) for stimulating discussions and insight. F.J.V. is supported by INCa 2019-125, E.B.C. thanks M. Dustin for support through ERC AdG 670930. D.R.F.C. is supported by the BBSRC (BB/P006205/1) and Cancer Research UK (A28052). V.H. and J.G.G. are funded by La Ligue contre le Cancer, by INCa (PLBIO19-291), by Plan Cancer (Nanotumor) and through institutional funds from University of Strasbourg and INSERM. K.R. is supported by the UEF Cell and Tissue Imaging Unit, Biocenter Kuopio and Biocenter Finland. We apologize to colleagues for any relevant work that could not be cited due to space restrictions.

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Author notes

  1. These authors contributed equally: Leonora Balaj, Chantal M. Boulanger, David R. F. Carter, Ewoud B. Compeer, Gisela D’Angelo, Samir El Andaloussi, Jacky G. Goetz, Julia C. Gross, Vincent Hyenne, Eva-Maria Krämer-Albers, Charles Lai, Xavier Loyer, Alex Marki, Stefan Momma, Esther Nolte ‘t Hoen, D. Michiel Pegtel, Hector Peinado, Graça Raposo, Kirsi Rilla, Hidetoshi Tahara, Clotilde Théry, Martin E. van Royen, Roosmarijn Vandenbroucke, Ann M. Wehman, Kenneth Witwer, Zhiwei Wu, Richard Wubbolts.

  2. These authors jointly supervised this work: Frederik J. Verweij, Guillaume van Niel.

Authors and Affiliations

  1. Université de Paris, Institute of Psychiatry and Neuroscience of Paris (IPNP), INSERM U1266, Paris, France

    Frederik J. Verweij & Guillaume van Niel

  2. GHU Paris Psychiatrie et Neurosciences, Hôpital Sainte Anne, Paris, France

    Frederik J. Verweij & Guillaume van Niel

  3. Department of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Leonora Balaj

  4. Université de Paris, PARCC, INSERM, Paris, France

    Chantal M. Boulanger & Xavier Loyer

  5. Department of Biological and Medical Sciences, Faculty of Health and Life Sciences, Oxford Brookes University, Oxford, UK

    David R. F. Carter

  6. Evox Therapeutics Limited, Oxford Science Park, Oxford, UK

    David R. F. Carter & Samir El Andaloussi

  7. Kennedy Institute of Rheumatology, NDORMS, University of Oxford, Oxford, UK

    Ewoud B. Compeer

  8. Institut Curie, PSL Research University, CNRS, UMR144 Cell Biology and Cancer, Paris, France

    Gisela D’Angelo & Graça Raposo

  9. Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden

    Samir El Andaloussi

  10. INSERM UMR_S1109, Tumor Biomechanics Lab, Université de Strasbourg, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Equipe Labellisée Ligue contre le Cancer, Strasbourg, France

    Jacky G. Goetz & Vincent Hyenne

  11. Health and Medical University, Potsdam, Germany

    Julia Christina Gross

  12. CNRS SNC5055, Strasbourg, France

    Vincent Hyenne

  13. Johannes Gutenberg-Universität Mainz, Institute of Developmental Biology and Neurobiology, Mainz, Germany

    Eva-Maria Krämer-Albers

  14. Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan

    Charles P. Lai

  15. La Jolla Institute for Allergy and Immunology, La Jolla, CA, USA

    Alex Marki

  16. Institute of Neurology (Edinger Institute), Goethe-University, Frankfurt am Main, Germany

    Stefan Momma

  17. Department of Biomolecular Health Sciences, Faculty of veterinary medicine, Utrecht University, Utrecht, the Netherlands

    Esther N. M. Nolte-‘t Hoen & Richard Wubbolts

  18. Amsterdam UMC, Vrije Universiteit Amsterdam, Pathology, Cancer Center Amsterdam, Amsterdam, the Netherlands

    D. Michiel Pegtel

  19. Microenvironment and Metastasis Laboratory, Molecular Oncology Programme, Spanish National Cancer Research Center (CNIO), Madrid, Spain

    Hector Peinado

  20. University of Eastern Finland, Institute of Biomedicine, Kuopio, Finland

    Kirsi Rilla

  21. Department of Cellular and Molecular Biology, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan

    Hidetoshi Tahara

  22. Institut Curie, PSL Research University, INSERM U932, Immunity and Cancer, Paris, France

    Clotilde Théry

  23. Erasmus MC, Department of Pathology, Rotterdam, the Netherlands

    Martin E. van Royen

  24. VIB Center for Inflammation Research and Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium

    Roosmarijn E. Vandenbroucke

  25. Department of Biological Sciences, University of Denver, Denver, CO, USA

    Ann M. Wehman

  26. Department of Molecular and Comparative Pathobiology and Neurology and the Richman Family Precision Medicine Center of Excellence in Alzheimer’s Disease, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Kenneth Witwer

  27. Center for Public Health Research, Medical School, Nanjing University, Nanjing, China

    Zhiwei Wu

  28. State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing, China

    Zhiwei Wu

  29. Medical School, Jiangsu Key Laboratory of Molecular Medicine, Nanjing University, Nanjing, China

    Zhiwei Wu

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Correspondence to Frederik J. Verweij or Guillaume van Niel.

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Competing interests

D.R.F.C. is employed by Evox Therapeutics. S.E.A. serves on the Scientific Advisory Board of EVOX Therapeutics. All other authors have no competing interests.

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Peer review information Rita Strack was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

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Verweij, F.J., Balaj, L., Boulanger, C.M. et al. The power of imaging to understand extracellular vesicle biology in vivo. Nat Methods 18, 1013–1026 (2021). https://doi.org/10.1038/s41592-021-01206-3

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