Targeting the adenosine 2A receptor enhances chimeric antigen receptor T cell efficacy
Paul A. Beavis, … , Michael H. Kershaw, Phillip K. Darcy
Paul A. Beavis, … , Michael H. Kershaw, Phillip K. Darcy
Published February 6, 2017
Citation Information: J Clin Invest. 2017;127(3):929-941. https://doi.org/10.1172/JCI89455.
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Research Article Immunology Oncology

Targeting the adenosine 2A receptor enhances chimeric antigen receptor T cell efficacy

Abstract

Chimeric antigen receptor (CAR) T cells have been highly successful in treating hematological malignancies, including acute and chronic lymphoblastic leukemia. However, treatment of solid tumors using CAR T cells has been largely unsuccessful to date, partly because of tumor-induced immunosuppressive mechanisms, including adenosine production. Previous studies have shown that adenosine generated by tumor cells potently inhibits endogenous antitumor T cell responses through activation of adenosine 2A receptors (A2ARs). Herein, we have observed that CAR activation resulted in increased A2AR expression and suppression of both murine and human CAR T cells. This was reversible using either A2AR antagonists or genetic targeting of A2AR using shRNA. In 2 syngeneic HER2+ self-antigen tumor models, we found that either genetic or pharmacological targeting of the A2AR profoundly increased CAR T cell efficacy, particularly when combined with PD-1 blockade. Mechanistically, this was associated with increased cytokine production of CD8+ CAR T cells and increased activation of both CD8+ and CD4+ CAR T cells. Given the known clinical relevance of the CD73/adenosine pathway in several solid tumor types, and the initiation of phase I trials for A2AR antagonists in oncology, this approach has high translational potential to enhance CAR T cell efficacy in several cancer types.

Authors

Paul A. Beavis, Melissa A. Henderson, Lauren Giuffrida, Jane K. Mills, Kevin Sek, Ryan S. Cross, Alexander J. Davenport, Liza B. John, Sherly Mardiana, Clare Y. Slaney, Ricky W. Johnstone, Joseph A. Trapani, John Stagg, Sherene Loi, Lev Kats, David Gyorki, Michael H. Kershaw, Phillip K. Darcy

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Figure 6

Targeting the A2AR with shRNA retroviral technology enhances CAR T cell function.

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Targeting the A2AR with shRNA retroviral technology enhances CAR T cell ...
CAR T cells were transduced with retroviruses encoding A2AR-directed shRNA or scrambled shRNA control and then selected with 2 μg/ml puromycin from day 4 to day 6. Eight days after activation, CAR T cells were then assessed for function. (A) 1 × 106 CAR T cells were reactivated with plate-bound anti-CD3/anti-CD28 (0.5 μg/ml) for 4 hours and then lysed for RNA analysis. Data are presented as the mean ± SD of triplicates and expressed relative to the production of IFN-γ of control cultures for each shRNA-transduced CAR T cell. (B) 1 × 105 CAR T cells were cocultured with 24JK/24JK-HER2 tumor cells at a 1:1 ratio in the presence or absence of NECA at indicated concentrations. Supernatants were collected after 16 hours and analyzed for concentration of IFN-γ. Data are presented relative to the IFN-γ production of control cultures and as the mean ± SD of quadruplicates from a representative experiment of n = 2. *P < 0.05, **P < 0.01 by 1-way ANOVA.