Adenosine mediates functional and metabolic suppression of peripheral and tumor-infiltrating CD8+ T cells. Issue 1 (10th October 2019)

• Record Type: Journal Article
• Authors: Mastelic-Gavillet, Beatris; Navarro Rodrigo, Blanca; Décombaz, Laure; Wang, Haiping; Ercolano, Giuseppe; Ahmed, Rita; Lozano, Leyder Elena; Ianaro, Angela; Derré, Laurent; Valerio, Massimo; Tawadros, Thomas; Jichlinski, Patrice; Nguyen-Ngoc, Tu; Speiser, Daniel E.; Verdeil, Grégory; Gestermann, Nicolas; Dormond, Olivier; Kandalaft, Lana; Coukos, George; Jandus, Camilla; Ménétrier-Caux, Christine; Caux, Christophe; Ho, Ping-Chih; Romero, Pedro; Harari, Alexandre; Vigano, Selena
• Is Part Of: Journal for immunotherapy of cancer. Volume 7:Issue 1(2019)
• Publication Date: 2019-10-10
• Subjects: Adenosine -- CD8 T cells -- Metabolism -- mTOR -- TILs; Cancer -- Immunotherapy -- Periodicals; Cancer -- Immunological aspects -- Periodicals; Tumors -- Immunological aspects -- Periodicals; Immunotherapy -- Periodicals; 616.99406105
• DOI: 10.1186/s40425-019-0719-5
• Languages: English
• ISSNs: 2051-1426

Abstract

Abstract : Background: Several mechanisms are present in the tumor microenvironment (TME) to impair cytotoxic T cell responses potentially able to control tumor growth. Among these, the accumulation of adenosine (Ado) contributes to tumor progression and represents a promising immunotherapeutic target. Ado has been shown to impair T cell effector function, but the role and mechanisms employed by Ado/Ado receptors (AdoRs) in modulating human peripheral and tumor-infiltrating lymphocyte (TIL) function are still puzzling. Methods: CD8 + T cell cytokine production following stimulation was quantified by intracellular staining and flow cytometry. The cytotoxic capacity of tumor infiltrating lymphocytes (TILs) was quantified by the chromium release assay following co-culture with autologous or anti-CD3-loaded tumor cell lines. The CD8 + T cell metabolic fitness was evaluated by the seahorse assay and by the quantification of 2-NBDG uptake and CD71/CD98 upregulation upon stimulation. The expression of AdoRs was assessed by RNA flow cytometry, a recently developed technology that we validated by semiquantitative RT-PCR (qRT-PCR), while the impact on T cell function was evaluated by the use of selective antagonists and agonists. The influence of Ado/AdoR on the PKA and mTOR pathways was evaluated by phosphoflow staining of p-CREB and p-S6, respectively, and validated by western blot. Results: Here, we demonstrate that Ado signaling through the A2A receptor (A2AR) in human peripheral CD8 + T cells and TILs is responsible for the higher sensitivity to Ado-mediated suppression of T central memory cells. We confirmed that Ado is able to impair peripheral and tumor-expanded T cell effector functions, and we show for the first time its impact on metabolic fitness. The Ado-mediated immunosuppressive effects are mediated by increased PKA activation that results in impairment of the mTORC1 pathway. Conclusions: Our findings unveil A2AR/PKA/mTORC1 as the main Ado signaling pathway impairing the immune competence of peripheral T cells and TILs. Thus, p-CREB and p-S6 may represent useful pharmacodynamic and efficacy biomarkers of immunotherapies targeting Ado. The effect of Ado on T cell metabolic fitness reinforces the importance of the adenosinergic pathway as a target for next-generation immunotherapy.