Adenosine 2A receptor and TIM3 suppress cytolytic killing of tumor cells via cytoskeletal polarization

Grace L Edmunds; Carissa C W Wong; Rachel Ambler; Emily J Milodowski; Hanin Alamir; Stephen J Cross; Gabriella Galea; Christoph Wülfing; David J Morgan

doi:10.1101/2021.05.20.444944v1

Adenosine 2A receptor and TIM3 suppress cytolytic killing of tumor cells via cytoskeletal polarization

Grace L Edmunds, Carissa C W Wong, Rachel Ambler, Emily J Milodowski, Hanin Alamir, Stephen J Cross, Gabriella Galea, Christoph Wülfing^*, David J Morgan^*

^*Corresponding author for this work

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Abstract

Tumors generate an immune-suppressive environment that prevents effective killing of tumor cells by CD8⁺ cytotoxic T cells (CTL). It remains largely unclear upon which cell type and at which stage of the anti-tumor response mediators of suppression act. We have combined an in vivo tumor model with a matching in vitro reconstruction of the tumor microenvironment based on tumor spheroids to identify suppressors of anti-tumor immunity that directly act on interaction between CTL and tumor cells and to determine mechanisms of action. An adenosine 2A receptor antagonist, as enhanced by blockade of TIM3, slowed tumor growth in vivo. Engagement of the adenosine 2A receptor and TIM3 reduced tumor cell killing in spheroids, impaired CTL cytoskeletal polarization ex vivo and in vitro and inhibited CTL infiltration into tumors and spheroids. With this role in CTL killing, blocking A_2AR and TIM3 may complement therapies that enhance T cell priming, e.g. anti-PD-1 and anti-CTLA-4.

Original language	English
Article number	9
Number of pages	17
Journal	Communications Biology
Volume	5
Issue number	1
Early online date	22 Jan 2022
DOIs	https://doi.org/10.1101/2021.05.20.444944v1 https://doi.org/10.1038/s42003-021-02972-8
Publication status	E-pub ahead of print - 22 Jan 2022

Bibliographical note

Funding Information:
We acknowledge the University of Bristol Flow Cytometry Facility with Andrew Herman and Lorena Sueiro-Ballesteros and the Wolfson BioImaging Facility with Katie Jepson for experimental support, Alan Hedges for statistical advice, and Mick Bailey for contributions to the principal component analysis. This work was supported by grants from the Wellcome Trust via the University of Bristol Elizabeth Blackwell Institute (Wellcome Trust ISSF2 grant 105612/Z/14/Z to G.L.E.), the Wellcome Trust (201254/Z/16/Z/ to G.L.E., 102387/Z/13/Z/ to R.A.) and the University of Bristol Cancer Research Fund (to C.W.). G.L.E was also supported as an associate of the Wellcome Trust GW4-CAT scheme. C.C.W.W. was supported by the MRC GW4 DTP. E.J.M. was supported by a Wellcome Trust GW4-CAT Fellowship. For the purpose of Open Access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission.

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© 2022, The Author(s).

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Edmunds et al.
Wuelfing, C. (Creator), University of Bristol, 20 May 2021
DOI: 10.5523/bris.11ocsor59owa32ihsxmf0qzj3s, http://data.bris.ac.uk/data/dataset/11ocsor59owa32ihsxmf0qzj3s
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Edmunds, G. L., Wong, C. C. W., Ambler, R., Milodowski, E. J., Alamir, H., Cross, S. J., Galea, G., Wülfing, C., & Morgan, D. J. (2022). Adenosine 2A receptor and TIM3 suppress cytolytic killing of tumor cells via cytoskeletal polarization. Communications Biology, 5(1), Article 9. Advance online publication. https://doi.org/10.1101/2021.05.20.444944v1, https://doi.org/10.1038/s42003-021-02972-8

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abstract = "Tumors generate an immune-suppressive environment that prevents effective killing of tumor cells by CD8+ cytotoxic T cells (CTL). It remains largely unclear upon which cell type and at which stage of the anti-tumor response mediators of suppression act. We have combined an in vivo tumor model with a matching in vitro reconstruction of the tumor microenvironment based on tumor spheroids to identify suppressors of anti-tumor immunity that directly act on interaction between CTL and tumor cells and to determine mechanisms of action. An adenosine 2A receptor antagonist, as enhanced by blockade of TIM3, slowed tumor growth in vivo. Engagement of the adenosine 2A receptor and TIM3 reduced tumor cell killing in spheroids, impaired CTL cytoskeletal polarization ex vivo and in vitro and inhibited CTL infiltration into tumors and spheroids. With this role in CTL killing, blocking A2AR and TIM3 may complement therapies that enhance T cell priming, e.g. anti-PD-1 and anti-CTLA-4.",

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AU - Cross, Stephen J

AU - Galea, Gabriella

AU - Wülfing, Christoph

AU - Morgan, David J

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N2 - Tumors generate an immune-suppressive environment that prevents effective killing of tumor cells by CD8+ cytotoxic T cells (CTL). It remains largely unclear upon which cell type and at which stage of the anti-tumor response mediators of suppression act. We have combined an in vivo tumor model with a matching in vitro reconstruction of the tumor microenvironment based on tumor spheroids to identify suppressors of anti-tumor immunity that directly act on interaction between CTL and tumor cells and to determine mechanisms of action. An adenosine 2A receptor antagonist, as enhanced by blockade of TIM3, slowed tumor growth in vivo. Engagement of the adenosine 2A receptor and TIM3 reduced tumor cell killing in spheroids, impaired CTL cytoskeletal polarization ex vivo and in vitro and inhibited CTL infiltration into tumors and spheroids. With this role in CTL killing, blocking A2AR and TIM3 may complement therapies that enhance T cell priming, e.g. anti-PD-1 and anti-CTLA-4.

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Adenosine 2A receptor and TIM3 suppress cytolytic killing of tumor cells via cytoskeletal polarization

Abstract

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