Ral GTPases promote breast cancer metastasis by controlling biogenesis and organ targeting of exosomes

  1. Shima Ghoroghi
  2. Benjamin Mary
  3. Annabel Larnicol
  4. Nandini Asokan
  5. Annick Klein
  6. Naël Osmani
  7. Ignacio Busnelli
  8. François Delalande
  9. Nicodème Paul
  10. Sébastien Halary
  11. Frédéric Gros
  12. Laetitia Fouillen
  13. Anne-Marie Haeberle
  14. Cathy Royer
  15. Coralie Spiegelhalter
  16. Gwennan André-Grégoire
  17. Vincent Mittelheisser
  18. Alexandre Detappe
  19. Kendelle Murphy
  20. Paul Timpson
  21. Raphaël Carapito
  22. Marcel Blot-Chabaud
  23. Julie Gavard
  24. Christine Carapito
  25. Nicolas Vitale
  26. Olivier Lefebvre
  27. Jacky G Goetz  Is a corresponding author
  28. Vincent Hyenne  Is a corresponding author
  1. INSERM U1109, France
  2. Laboratoire de Spectrométrie de Masse BioOrganique, France
  3. Muséum National d'Histoire Naturelle de Paris, France
  4. Université de Bordeaux, CNRS, France
  5. Centre National de la Recherche Scientifique, Université de Strasbourg, France
  6. CNRS UPS 3156, France
  7. IGBMC (UMR7104)/ INSERM (U1258)/ Université de Strasbourg,, France
  8. CRCINA, INSERM, CNRS, Université de Nantes, Université d'Angers, France
  9. Icans, France
  10. Garvan Institute of Medical Research, Australia
  11. INSERM 1263, Inrae 1260, France
  12. Institut des Neurosciences Cellulaires et Intégratives; UPR-3212 CNRS, Strasbourg University, France

Abstract

Cancer extracellular vesicles (EVs) shuttle at distance and fertilize pre-metastatic niches facilitating subsequent seeding by tumor cells. However, the link between EV secretion mechanisms and their capacity to form pre-metastatic niches remains obscure. Using mouse models, we show that GTPases of the Ral family control, through the phospholipase D1, multi-vesicular bodies homeostasis and tune the biogenesis and secretion of pro-metastatic EVs. Importantly, EVs from RalA or RalB depleted cells have limited organotropic capacities in vivo and are less efficient in promoting metastasis. RalA and RalB reduce the EV levels of the adhesion molecule MCAM/CD146, which favors EV-mediated metastasis by allowing EVs targeting to the lungs. Finally, RalA, RalB and MCAM/CD146, are factors of poor prognosis in breast cancer patients. Altogether, our study identifies RalGTPases as central molecules linking the mechanisms of EVs secretion and cargo loading to their capacity to disseminate and induce pre-metastatic niches in a CD146 dependent manner.

Data availability

Sequencing and mass spectrometry data have been deposited to EV-Track knowledgebase

The following previously published data sets were used

Article and author information

Author details

  1. Shima Ghoroghi

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Benjamin Mary

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Annabel Larnicol

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  4. Nandini Asokan

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  5. Annick Klein

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  6. Naël Osmani

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Ignacio Busnelli

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  8. François Delalande

    IPHC UMR 7178, Laboratoire de Spectrométrie de Masse BioOrganique, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  9. Nicodème Paul

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4680-3012
  10. Sébastien Halary

    CNRS, UMR 7245 MCAM, Muséum National d'Histoire Naturelle de Paris, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  11. Frédéric Gros

    Lymphocyte Homeostasis and autoimmunity, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6252-4323
  12. Laetitia Fouillen

    Laboratoire de Biogenèse Membranaire, UMR 5200, Université de Bordeaux, CNRS, Villenave d'Ornon, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1204-9296
  13. Anne-Marie Haeberle

    Institut des Neurosciences Cellulaires et Intégratives, Centre National de la Recherche Scientifique, Université de Strasbourg, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  14. Cathy Royer

    Plateforme Imagerie In Vitro, CNRS UPS 3156, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  15. Coralie Spiegelhalter

    Imaging Center, IGBMC (UMR7104)/ INSERM (U1258)/ Université de Strasbourg,, Illkirch, France
    Competing interests
    The authors declare that no competing interests exist.
  16. Gwennan André-Grégoire

    Team Soap, CRCINA, INSERM, CNRS, Université de Nantes, Université d'Angers, Nantes, France
    Competing interests
    The authors declare that no competing interests exist.
  17. Vincent Mittelheisser

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  18. Alexandre Detappe

    Nanotranslational laboratory, Icans, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  19. Kendelle Murphy

    The Kinghorn Cancer Centre, Garvan Institute of Medical Research, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  20. Paul Timpson

    Cancer Division, Garvan Institute of Medical Research, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  21. Raphaël Carapito

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  22. Marcel Blot-Chabaud

    C2VN, INSERM 1263, Inrae 1260, Marseille, France
    Competing interests
    The authors declare that no competing interests exist.
  23. Julie Gavard

    Team Soap, CRCINA, INSERM, CNRS, Université de Nantes, Université d'Angers, Nantes, France
    Competing interests
    The authors declare that no competing interests exist.
  24. Christine Carapito

    IPHC UMR 7178, Laboratoire de Spectrométrie de Masse BioOrganique, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  25. Nicolas Vitale

    Neuronal communication and networks, Institut des Neurosciences Cellulaires et Intégratives; UPR-3212 CNRS, Strasbourg University, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  26. Olivier Lefebvre

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.
  27. Jacky G Goetz

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    For correspondence
    jacky.goetz@inserm.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2842-8116
  28. Vincent Hyenne

    Tumor Biomechanics, INSERM U1109, Strasbourg, France
    For correspondence
    hyenne@unistra.fr
    Competing interests
    The authors declare that no competing interests exist.

Funding

Institut National Du Cancer (PLBIO19-291)

  • Jacky G Goetz

La Ligue contre le cancer

  • Jacky G Goetz

Canceropole grand est

  • Jacky G Goetz

Plan Cancer

  • Jacky G Goetz

Roche

  • Jacky G Goetz

Agence Nationale de la Recherche (ANR-10-INBS-08-03)

  • Christine Carapito

Agence Nationale de la Recherche (ANR-19-CE44-0019)

  • Nicolas Vitale

Agence Nationale de la Recherche (ANR-11-INBS- 0010)

  • Laetitia Fouillen

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animals were housed and han- dled according to the guidelines of INSERM and the eth- ical committee of Alsace, France (CREMEAS) (Direc- tive 2010/63/EU on the protection of animals used for scientific purposes). Animal facility agreement num- ber: C67-482-33. Experimental license for mice: Apafis 4707-20l6032416407780; experimental license for zebrafish: Apafis 16862-2018121914292754.

Human subjects: Paraffin sections of 4 µm from metastasic and non-metastasic breast tumours were obtained from CRB-Tumorothèque of the Institut de Cancérologie de l'Ouest (ICO, Saint-Herblain, France) (Heymann et al., 2020). Patients were diagnosed and treated at ICO (Integrated Center for Oncology, St Herblain) for metastatic and non-metastatic breast cancers. Samples were processed and included in the CRB-Tumorothèque ICO upon donor agreement and informed consent. Samples and related information are destroyed at the request of the donor. The CRB-Tumorothèque ICO have been declared to and authorized by the French Research Ministry (Declaration Number: DC-2018-3321). This declaration includes approval by a research ethics committee (CPP "Comité de protection des personnes"), in accordance with the French legislation of the Public Health Code.

Reviewing Editor

  1. Erica A Golemis, Fox Chase Cancer Center, United States

Publication history

  1. Received: July 28, 2020
  2. Accepted: January 5, 2021
  3. Accepted Manuscript published: January 6, 2021 (version 1)
  4. Version of Record published: January 22, 2021 (version 2)

Copyright

© 2021, Ghoroghi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Shima Ghoroghi
  2. Benjamin Mary
  3. Annabel Larnicol
  4. Nandini Asokan
  5. Annick Klein
  6. Naël Osmani
  7. Ignacio Busnelli
  8. François Delalande
  9. Nicodème Paul
  10. Sébastien Halary
  11. Frédéric Gros
  12. Laetitia Fouillen
  13. Anne-Marie Haeberle
  14. Cathy Royer
  15. Coralie Spiegelhalter
  16. Gwennan André-Grégoire
  17. Vincent Mittelheisser
  18. Alexandre Detappe
  19. Kendelle Murphy
  20. Paul Timpson
  21. Raphaël Carapito
  22. Marcel Blot-Chabaud
  23. Julie Gavard
  24. Christine Carapito
  25. Nicolas Vitale
  26. Olivier Lefebvre
  27. Jacky G Goetz
  28. Vincent Hyenne
(2021)
Ral GTPases promote breast cancer metastasis by controlling biogenesis and organ targeting of exosomes
eLife 10:e61539.
https://doi.org/10.7554/eLife.61539

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