Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Extracellular vesicles are independent metabolic units with asparaginase activity

Nature Chemical Biology volume 13, pages 951–955 (2017)Cite this article

Subjects

Abstract

Extracellular vesicles (EVs) are membrane particles involved in the exchange of a broad range of bioactive molecules between cells and the microenvironment. Although it has been shown that cells can traffic metabolic enzymes via EVs, much remains to be elucidated with regard to their intrinsic metabolic activity. Accordingly, herein we assessed the ability of neural stem/progenitor cell (NSC)-derived EVs to consume and produce metabolites. Our metabolomics and functional analyses both revealed that EVs harbor L-asparaginase activity, catalyzed by the enzyme asparaginase-like protein 1 (Asrgl1). Critically, we show that Asrgl1 activity is selective for asparagine and is devoid of glutaminase activity. We found that mouse and human NSC EVs traffic Asrgl1. Our results demonstrate, for the first time, that NSC EVs function as independent metabolic units that are able to modify the concentrations of critical nutrients, with the potential to affect the physiology of their microenvironment.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: NSCs secrete EVs containing exosomes.
Figure 2: NSC EVs are metabolically active in vitro.
Figure 3: Mouse NSCs traffic Asrgl1 into EVs.
Figure 4: Asrgl1 is responsible for the selective L-asparaginase activity associated with EVs.

Similar content being viewed by others

References

  1. 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).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  3. Kim, D.K., Lee, J., Simpson, R.J., Lötvall, J. & Gho, Y.S. EVpedia: A community web resource for prokaryotic and eukaryotic extracellular vesicles research. Semin. Cell Dev. Biol. 40, 4–7 (2015).

    Article  CAS  Google Scholar 

  4. Kalra, H. et al. Vesiclepedia: a compendium for extracellular vesicles with continuous community annotation. PLoS Biol. 10, e1001450 (2012).

    Article  CAS  Google Scholar 

  5. Théry, C., Ostrowski, M. & Segura, E. Membrane vesicles as conveyors of immune responses. Nat. Rev. Immunol. 9, 581–593 (2009).

    Article  Google Scholar 

  6. Cossetti, C. et al. Extracellular vesicles from neural stem cells transfer IFN-γ via Ifngr1 to activate Stat1 signaling in target cells. Mol. Cell 56, 193–204 (2014).

    Article  CAS  Google Scholar 

  7. Garcia, N.A., Moncayo-Arlandi, J., Sepulveda, P. & Diez-Juan, A. Cardiomyocyte exosomes regulate glycolytic flux in endothelium by direct transfer of GLUT transporters and glycolytic enzymes. Cardiovasc. Res. 109, 397–408 (2016).

    Article  CAS  Google Scholar 

  8. Ronquist, K.G., Ek, B., Stavreus-Evers, A., Larsson, A. & Ronquist, G. Human prostasomes express glycolytic enzymes with capacity for ATP production. Am. J. Physiol. Endocrinol. Metab. 304, E576–E582 (2013).

    Article  CAS  Google Scholar 

  9. Minciacchi, V.R. et al. Large oncosomes contain distinct protein cargo and represent a separate functional class of tumor-derived extracellular vesicles. Oncotarget 6, 11327–11341 (2015).

    Article  Google Scholar 

  10. Johnson, S.M. et al. Metabolic reprogramming of bone marrow stromal cells by leukemic extracellular vesicles in acute lymphoblastic leukemia. Blood 128, 453–456 (2016).

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  12. Lobb, R.J. et al. Optimized exosome isolation protocol for cell culture supernatant and human plasma. J. Extracell. Vesicles 4, 27031 (2015).

    Article  Google Scholar 

  13. Théry, C. et al. Molecular characterization of dendritic cell-derived exosomes. Selective accumulation of the heat shock protein hsc73. J. Cell Biol. 147, 599–610 (1999).

    Article  Google Scholar 

  14. Quillin, M.L. & Matthews, B.W. Accurate calculation of the density of proteins. Acta Crystallogr. D Biol. Crystallogr. 56, 791–794 (2000).

    Article  CAS  Google Scholar 

  15. Mazzini, L. et al. Human neural stem cell transplantation in ALS: initial results from a phase I trial. J. Transl. Med. 13, 17 (2015).

    Article  Google Scholar 

  16. Poliakov, A., Spilman, M., Dokland, T., Amling, C.L. & Mobley, J.A. Structural heterogeneity and protein composition of exosome-like vesicles (prostasomes) in human semen. Prostate 69, 159–167 (2009).

    Article  Google Scholar 

  17. Cantor, J.R., Stone, E.M., Chantranupong, L. & Georgiou, G. The human asparaginase-like protein 1 hASRGL1 is an Ntn hydrolase with β-aspartyl peptidase activity. Biochemistry 48, 11026–11031 (2009).

    Article  CAS  Google Scholar 

  18. Kourembanas, S. Exosomes: vehicles of intercellular signaling, biomarkers, and vectors of cell therapy. Annu. Rev. Physiol. 77, 13–27 (2015).

    Article  CAS  Google Scholar 

  19. Tkach, M. & Théry, C. Communication by extracellular vesicles: where we are and where we need to go. Cell 164, 1226–1232 (2016).

    Article  CAS  Google Scholar 

  20. Xu, L., Yang, B.F. & Ai, J. MicroRNA transport: a new way in cell communication. J. Cell. Physiol. 228, 1713–1719 (2013).

    Article  CAS  Google Scholar 

  21. Fafournoux, P., Bruhat, A. & Jousse, C. Amino acid regulation of gene expression. Biochem. J. 351, 1–12 (2000).

    Article  CAS  Google Scholar 

  22. Braicu, C. et al. Exosomes as divine messengers: are they the Hermes of modern molecular oncology? Cell Death Differ. 22, 34–45 (2015).

    Article  CAS  Google Scholar 

  23. Christie, G.R., Hyde, R. & Hundal, H.S. Regulation of amino acid transporters by amino acid availability. Curr. Opin. Clin. Nutr. Metab. Care 4, 425–431 (2001).

    Article  CAS  Google Scholar 

  24. Witwer, K.W. et al. Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J. Extracell. Vesicles 2, (2013).

  25. Birsoy, K. et al. An essential role of the mitochondrial electron transport chain in cell proliferation is to enable aspartate synthesis. Cell 162, 540–551 (2015).

    Article  CAS  Google Scholar 

  26. Sullivan, L.B. et al. Supporting aspartate biosynthesis is an essential function of respiration in proliferating cells. Cell 162, 552–563 (2015).

    Article  CAS  Google Scholar 

  27. Petroff, O.A. GABA and glutamate in the human brain. Neuroscientist 8, 562–573 (2002).

    Article  CAS  Google Scholar 

  28. Watanabe, M. et al. Gamma-aminobutyric acid (GABA) and cell proliferation: focus on cancer cells. Histol. Histopathol. 21, 1135–1141 (2006).

    CAS  PubMed  Google Scholar 

  29. Stepulak, A., Rola, R., Polberg, K. & Ikonomidou, C. Glutamate and its receptors in cancer. J. Neural Transm. (Vienna) 121, 933–944 (2014).

    Article  CAS  Google Scholar 

  30. Pavlides, S. et al. The reverse Warburg effect: aerobic glycolysis in cancer associated fibroblasts and the tumor stroma. Cell Cycle 8, 3984–4001 (2009).

    Article  CAS  Google Scholar 

  31. Sonveaux, P. et al. Targeting the lactate transporter MCT1 in endothelial cells inhibits lactate-induced HIF-1 activation and tumor angiogenesis. PLoS One 7, e33418 (2012).

    Article  CAS  Google Scholar 

  32. Haas, R. et al. Lactate regulates metabolic and pro-inflammatory circuits in control of T cell migration and effector functions. PLoS Biol. 13, e1002202 (2015).

    Article  Google Scholar 

  33. Bush, L.A. et al. A novel asparaginase-like protein is a sperm autoantigen in rats. Mol. Reprod. Dev. 62, 233–247 (2002).

    Article  CAS  Google Scholar 

  34. Duval, M. et al. Comparison of Escherichia coli-asparaginase with Erwinia-asparaginase in the treatment of childhood lymphoid malignancies: results of a randomized European Organisation for Research and Treatment of Cancer-Children's Leukemia Group phase 3 trial. Blood 99, 2734–2739 (2002).

    Article  CAS  Google Scholar 

  35. Karpel-Massler, G. et al. Metabolic reprogramming of glioblastoma cells by L-asparaginase sensitizes for apoptosis in vitro and in vivo. Oncotarget 7, 33512–33528 (2016).

    PubMed  PubMed Central  Google Scholar 

  36. Ollenschläger, G. et al. Asparaginase-induced derangements of glutamine metabolism: the pathogenetic basis for some drug-related side-effects. Eur. J. Clin. Invest. 18, 512–516 (1988).

    Article  Google Scholar 

  37. Iraci, N., Tyzack, G.E., Cossetti, C., Alfaro-Cervello, C. & Pluchino, S. Viral manipulation of neural stem/precursor cells. Neuromethods 82, 269–288 (2013).

    Article  Google Scholar 

Download references

Acknowledgements

The authors thank F. Dazzi, C. Mauro, J. Smith and A. Tolkovsky for critically discussing the article. We are grateful to J. Muzard (iZON) for his help with the qNano. We acknowledge the technical assistance of I. Bicci, M. Davis, F. Gessler, G. Pluchino and B. Vega-Blanco. This work has received support from the Italian Multiple Sclerosis Association (AISM; grant 2010/R/31 and grant 2014/PMS/4 to S.P.); the Italian Ministry of Health (GR08-7 to S.P.); the European Research Council (ERC) under the ERC-2010-StG Grant agreement no. 260511-SEM_SEM; the Medical Research Council, the Engineering and Physical Sciences Research Council and the Biotechnology and Biological Sciences Research Council UK Regenerative Medicine Platform Hub “Acellular Approaches for Therapeutic Delivery” (MR/K026682/1 to S.P.); The Evelyn Trust (RG 69865 to S.P.); The Bascule Charitable Trust (RG 75149 to S.P.); and core support grant from the Wellcome Trust and Medical Research Council to the Wellcome Trust–MRC Cambridge Stem Cell Institute. N.I. was supported by a FEBS long-term fellowship. C.F., A.S.H., and E.G. were funded by the Medical Research Council, Core Fund SKAG006.

Author information

Author notes

  1. Nunzio Iraci

    Present address: Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, Catania, Italy

  2. Nunzio Iraci and Edoardo Gaude: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Clinical Neurosciences–Division of Stem Cell Neurobiology, Wellcome Trust–Medical Research Council Stem Cell Institute, and NIHR Biomedical Research Centre, University of Cambridge, Cambridge, UK

    Nunzio Iraci, Tommaso Leonardi, Chiara Cossetti, Luca Peruzzotti-Jametti, Joshua D Bernstock & Stefano Pluchino

  2. MRC Cancer Unit, University of Cambridge, Hutchison/MRC Research Centre, Cambridge Biomedical Campus, Cambridge, UK

    Edoardo Gaude, Ana S H Costa & Christian Frezza

  3. European Molecular Biology Laboratory, European Bioinformatics Institute (EMBL-EBI), Wellcome Trust Genome Campus, Hinxton, Cambridge, UK

    Tommaso Leonardi, Harpreet K Saini & Anton J Enright

  4. Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health (NINDS/NIH), Bethesda, Maryland, USA

    Joshua D Bernstock

  5. Stem Cells Laboratory, Cell Factory and Biobank, Azienda Ospedaliera 'Santa Maria', Terni, Italy

    Maurizio Gelati

  6. IRCCS Casa Sollievo della Sofferenza, Foggia, Italy

    Maurizio Gelati & Angelo Luigi Vescovi

  7. Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milano, Italy

    Angelo Luigi Vescovi

  8. Medical Research Council–Elsie Widdowson Laboratory, Cambridge, UK

    Carlos Bastos & Nuno Faria

  9. Department of Engineering, Electrical Engineering Division, University of Cambridge, Cambridge, UK

    Luigi G Occhipinti

Authors
  1. Nunzio Iraci
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Edoardo Gaude
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tommaso Leonardi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ana S H Costa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chiara Cossetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Luca Peruzzotti-Jametti
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Joshua D Bernstock
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Harpreet K Saini
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Maurizio Gelati
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Angelo Luigi Vescovi
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Carlos Bastos
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Nuno Faria
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Luigi G Occhipinti
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Anton J Enright
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Christian Frezza
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Stefano Pluchino
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

N.I., E.G., C.F. and S.P. conceived the study and designed the experiments; N.I., T.L., C.C., L.P.-J., J.D.B. and M.G. elucidated the trafficking of Asrgl1 into NSC EVs, and performed experiments including cell culture preparations, EV purification from media, tunable sensitive pulse sensing, western blots, RT and qPCR, vector production and generation of gain-of-function and loss-of-function tools; N.I., C.B. and N.F. performed nanoparticle tracking analysis; A.S.H.C. and E.G. performed the LC–MS metabolomic analyses; A.L.V., L.G.O., C.F. and S.P. provided key reagents and resources; N.I., E.G., A.S.H.C., T.L., H.K.S., A.J.E., C.F. and S.P. analyzed the data and interpreted and discussed the results; N.I., E.G., C.F. and S.P. prepared the figures and wrote and edited the manuscript; C.F. and S.P. supervised the research.

Corresponding authors

Correspondence to Christian Frezza or Stefano Pluchino.

Ethics declarations

Competing interests

N.I., E.G., T.L., C.F. and S.P. are listed as inventors on a patent application related to the technology described in this work (European Patent application no. 16189525.5).

Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Table 1 and Supplementary Figures 1–12 (PDF 2598 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Iraci, N., Gaude, E., Leonardi, T. et al. Extracellular vesicles are independent metabolic units with asparaginase activity. Nat Chem Biol 13, 951–955 (2017). https://doi.org/10.1038/nchembio.2422

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nchembio.2422

This article is cited by

Search

Advanced search

Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research