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Tumoral P2Y2 receptor modulates tumor growth and host anti-tumor immune responses in a syngeneic murine model of oral cancer

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Abstract

Head and neck squamous cell carcinomas (HNSCCs) are a heterogenous group of tumors and among the top 10 most common cancers and they arise from the epithelial tissues of the mucosal surfaces of the oral cavity, oropharynx, and larynx. Aberrant purinergic signaling has been associated with various cancer types. Here, we studied the role of the P2Y2 purinergic receptor (P2Y2R) in the context of oral cancer. We utilized bioinformatics analysis of deposited datasets to examine purinome gene expression in HNSCC tumors and cells lines and functionally characterized nucleotide-induced P2 receptor signaling in human FaDu and Cal27 and murine MOC2 oral cancer cell lines. Utilizing tumorigenesis assays with wild-type or P2ry2 knockout MOC2 cells we evaluated the role of P2Y2Rs in tumor growth and the host anti-tumor immune responses. Our data demonstrate that human and murine oral cancer cell lines express numerous P2 receptors, with the P2Y2R being highly expressed. Using syngeneic tumor grafts in wild-type mice, we observed that MOC2 tumors expressing P2Y2R were larger than P2Y2R−/− tumors. Wild-type MOC2 tumors contained a lower population of tumor-infiltrating CD11b+F4/80+ macrophages and CD3+ cells, which were revealed to be CD3+CD4+IFNγ+ T cells, compared to P2Y2R−/− tumors. These results were mirrored when utilizing P2Y2R−/− mice, indicating that the changes in MOC2 tumor growth and to the host anti-tumor immune response were independent of host derived P2Y2Rs. Results suggest that targeted suppression of the P2Y2R in HNSCC cells in vivo, rather than systemic P2Y2R antagonism, may be a more effective treatment strategy for HNSCCs.

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Data availability

The flow cytometry data underlying this article are available as .fcs files deposited at FLOWRepository under ID: FR-FCM-Z5H3. All other data underlying this article will be shared on reasonable request to the corresponding author.

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Acknowledgements

The authors would like to thank Mr. Daniel Jackson (University of Missouri, Cell and Immunobiology Core) and Dr. Joel Crespo (Cytek) for their assistance and consultation with spectral flow cytometry.

Funding

This work was supported by grant R01-DE007389 from the National Institute of Dental and Craniofacial Research and a TRIUMPH grant from the University of Missouri School of Medicine. KMF was supported by a Life Sciences Fellowship and The Wayne L. Ryan Fellowship from the University of Missouri and The Ryan Foundation, respectively.

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Authors and Affiliations

  1. Department of Biochemistry, University of Missouri, Columbia, MO, USA

    Kevin Muñoz Forti, Lucas T. Woods, Kimberly J. Jasmer, Jean M. Camden & Gary A. Weisman

  2. Christopher S. Bond Life Sciences Center, University of Missouri, Columbia, MO, USA

    Kevin Muñoz Forti, Lucas T. Woods, Kimberly J. Jasmer, Jean M. Camden & Gary A. Weisman

Authors
  1. Kevin Muñoz Forti
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  2. Lucas T. Woods
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Contributions

KMF, LTW, JMC, and KJJ were responsible for the experimental design. LTW was responsible for bioinformatic analysis. KMF was responsible for cell culture and animal experiments. KMF and KJJ were responsible for flow cytometry analysis. KMF and LTW were responsible for data curation and generating figures. GAW was responsible for securing funding and supervising data collection. All authors contributed to the writing and review of the manuscript.

Corresponding author

Correspondence to Gary A. Weisman.

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All animal studies conducted in this work were reviewed and approved by the University of Missouri’s Animal Care and Use Committee (ACUC).

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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Forti, K.M., Woods, L.T., Jasmer, K.J. et al. Tumoral P2Y2 receptor modulates tumor growth and host anti-tumor immune responses in a syngeneic murine model of oral cancer. Purinergic Signalling (2023). https://doi.org/10.1007/s11302-023-09960-z

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