Abstract 4517: Targeting Notch1 via adenosine A2A receptor to modulate tumor immunity
Authors: Monticone Giulia , Hossain Fokhrul M. , Ucar Deniz A. , Majumder Samarpan , Sorrentino Claudia , Rodriguez Paulo C. , Sierra Rosa A. , Pannuti Antonio , Hatfield Stephen , Osborne Barbara A. , Minter Lisa M. , Morello Silvana , Miele Lucio 
Published: September 2020

Abstract
One of the hallmarks of cancer is the ability to evade the host immune system and this is achieved through different mechanisms. Several studies have showed that the accumulation of adenosine in the tumor microenvironment suppresses T-cell functions, thus helping the tumor to evade the immune system. Finding a way to counteract adenosine-mediated immune suppression might greatly enhance the host endogenous immune response against cancer and the efficacy of adoptive immunotherapies. To achieve this, we need to understand how adenosine regulates T-cell functions. In a collaborative project, our group recently showed that stimulation of adenosine A2A receptor (A2AR) reduces T-cell receptor (TCR)-signaling and consequently Notch1 activation and expression in CD8+ T-cells, which are critical to cancer immunity. This suggests that A2AR stimulation suppresses CD8+ T-cell function through inhibition of TCR-induced Notch1, which is required for the activation and function of CD8+ T-cells. Based on these observations, we hypothesized that while stimulation of A2AR suppresses CD8+ T-cells, inhibition of A2AR should protect CD8+ T-cells from the adenosine-mediated immune suppression. Notch1 is likely to be critical in this process since it was shown that ectopic expression of Notch1 intracellular domain prevents the adenosine-mediated immune suppression in CD8+ T-cells. Therefore, we aim to investigate the effect of A2AR inhibition on TCR activation and Notch1 to evaluate the A2AR-Notch axis as a novel immunotherapeutic target. Our data show that pharmacological inhibition of A2AR with a selective antagonist induces tumor cell death and increases the number of CD8+ T-cells in tumor-derived spheroids from a mouse triple-negative breast cancer (TNBC) model. The effect of A2AR inhibition appears to be immune-mediated since the same agent did not induce cell death in tumor-derived spheroids from immunocompromised mice. Along the same lines, we show that, inhibition of A2AR restored Notch1 activation and proliferation in primary murine CD8+ T-cells, but did not affect Notch1 and proliferation in TNBC cell lines. This effect is likely to be strictly dependent on Notch since A2AR inhibition failed to rescue CD8+ T-cell proliferation from the suppressive effect of gamma-secretase inhibition. It is not yet known how A2AR regulates Notch1, however, our latest preliminary results suggest that A2AR stimulation might promote the endosomal degradation of Notch1, whereas, A2AR inhibition might switch Notch1 fate from endosomal degradation to activation. Further investigation is needed to establish how A2AR regulates Notch1 and whether A2AR inhibition can protect CD8+ T-cell function in vivo in the tumor microenvironment. Overall our current data provides a rationale for the evaluation of A2AR antagonists as a Notch-modulating immunotherapy.
Citation Format: Giulia Monticone, Fokhrul M. Hossain, Deniz A. Ucar, Samarpan Majumder, Claudia Sorrentino, Paulo C. Rodriguez, Rosa A. Sierra, Antonio Pannuti, Stephen Hatfield, Barbara A. Osborne, Lisa M. Minter, Silvana Morello, Lucio Miele. Targeting Notch1 via adenosine A2A receptor to modulate tumor immunity [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4517.