Adenosine mediates functional and metabolic suppression of peripheral and tumor-infiltrating CD8+ T cells.
- Mastelic-Gavillet B Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Navarro Rodrigo B Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Décombaz L Department of Oncology, University of Lausanne, Lausanne, Switzerland.
- Wang H Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Ercolano G Department of Pharmacy, University of Naples Federico II, Naples, Italy.
- Ahmed R Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Lozano LE Department of Oncology, University of Lausanne, Lausanne, Switzerland.
- Ianaro A Department of Pharmacy, University of Naples Federico II, Naples, Italy.
- Derré L Department of Urology, Urology Research Unit, CHUV, Lausanne, Switzerland.
- Valerio M Department of Urology, Urology Research Unit, CHUV, Lausanne, Switzerland.
- Tawadros T Department of Urology, Urology Research Unit, CHUV, Lausanne, Switzerland.
- Jichlinski P Department of Urology, Urology Research Unit, CHUV, Lausanne, Switzerland.
- Nguyen-Ngoc T Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Speiser DE Department of Oncology, University of Lausanne, Lausanne, Switzerland.
- Verdeil G Department of Oncology, University of Lausanne, Lausanne, Switzerland.
- Gestermann N Department of Oncology, University of Lausanne, Lausanne, Switzerland.
- Dormond O Department of Visceral Surgery, CHUV, Lausanne, Switzerland.
- Kandalaft L Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Coukos G Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Jandus C Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Ménétrier-Caux C Department of Immunology Virology and Inflammation, Univ Lyon, Université Claude Bernard Lyon 1, 69008, Lyon, France.
- Caux C Department of Immunology Virology and Inflammation, Univ Lyon, Université Claude Bernard Lyon 1, 69008, Lyon, France.
- Ho PC Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Romero P Department of Oncology, University of Lausanne, Lausanne, Switzerland.
- Harari A Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
- Vigano S Department of Oncology, Ludwig Institute for Cancer Research Lausanne, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland. selena.vigano@chuv.ch.
- 2019-10-12
Published in:
- Journal for immunotherapy of cancer. - 2019
Adenosine
CD8 T cells
Metabolism
TILs
mTOR
Adenosine
Adult
Aged
Aged, 80 and over
CD8-Positive T-Lymphocytes
Cell Line, Tumor
Cyclic AMP-Dependent Protein Kinases
Disease Progression
Female
Humans
Lymphocytes, Tumor-Infiltrating
Male
Mechanistic Target of Rapamycin Complex 1
Middle Aged
Neoplasms
Primary Cell Culture
Receptor, Adenosine A2A
Signal Transduction
Tumor Escape
Tumor Microenvironment
English
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.
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.
- Language
-
- English
- Open access status
- gold
- Identifiers
-
- DOI 10.1186/s40425-019-0719-5
- PMID 31601268
- Persistent URL
- https://sonar.ch/global/documents/854
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