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Adenosine receptors regulate exosome production

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Abstract

Exosomes, small-sized extracellular vesicles, carry components of the purinergic pathway. The production by cells of exosomes carrying this pathway remains poorly understood. Here, we asked whether type 1, 2A, or 2B adenosine receptors (A1Rs, A2ARs, and A2BRs, respectively) expressed by producer cells are involved in regulating exosome production. Preglomerular vascular smooth muscle cells (PGVSMCs) were isolated from wildtype, A1R−/−, A2AR−/−, and A2BR−/− rats, and exosome production was quantified under normal or metabolic stress conditions. Exosome production was also measured in various cancer cells treated with pharmacologic agonists/antagonists of A1Rs, A2ARs, and A2BRs in the presence or absence of metabolic stress or cisplatin. Functional activity of exosomes was determined in Jurkat cell apoptosis assays. In PGVSMCs, A1R and A2AR constrained exosome production under normal conditions (p = 0.0297; p = 0.0409, respectively), and A1R, A2AR, and A2BR constrained exosome production under metabolic stress conditions. Exosome production from cancer cells was reduced (p = 0.0028) by the selective A2AR agonist CGS 21680. These exosomes induced higher levels of Jurkat apoptosis than exosomes from untreated cells or cells treated with A1R and A2BR agonists (p = 0.0474). The selective A2AR antagonist SCH 442416 stimulated exosome production under metabolic stress or cisplatin treatment, whereas the selective A2BR antagonist MRS 1754 reduced exosome production. Our findings indicate that A2ARs suppress exosome release in all cell types examined, whereas effects of A1Rs and A2BRs are dependent on cell type and conditions. Pharmacologic targeting of cancer with A2AR antagonists may inadvertently increase exosome production from tumor cells.

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Funding

N.L. was supported by the Leopoldina Fellowships LPDS 2017-12 and LPDR 2019-02 from German National Academy of Sciences Leopoldina. This work was supported by National Institutes of Health grants HL109002, DK091190, HL069846, DK068575, and DK079307 (to EKJ) and the NIH grant R01-CA 1686288 to TLW.

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Authors and Affiliations

  1. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213, USA

    Nils Ludwig, Juliana H. Azambuja & Theresa L. Whiteside

  2. UPMC Hillman Cancer Center, UPCI Research Pavilion, Suite 1.27, 5117 Centre Avenue, Pittsburgh, PA, 15213, USA

    Nils Ludwig, Juliana H. Azambuja, Aparna Rao & Theresa L. Whiteside

  3. Programa de Pós-Graduação em Biociências, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA), Porto Alegre, RS, Brazil

    Juliana H. Azambuja

  4. Department of Neurological Surgery, University of Pittsburgh Medical Center, Pittsburgh, PA, USA

    Aparna Rao

  5. Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Delbert G. Gillespie & Edwin K. Jackson

  6. Department of Immunology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213, USA

    Theresa L. Whiteside

  7. Department of Otolaryngology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15213, USA

    Theresa L. Whiteside

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  1. Nils Ludwig
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Correspondence to Theresa L. Whiteside.

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Ludwig, N., Azambuja, J.H., Rao, A. et al. Adenosine receptors regulate exosome production. Purinergic Signalling 16, 231–240 (2020). https://doi.org/10.1007/s11302-020-09700-7

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  • DOI: https://doi.org/10.1007/s11302-020-09700-7

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