CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy
Lauren Giuffrida, Kevin Sek, Melissa A. Henderson, Junyun Lai, Amanda X. Y. Chen, Déborah Meyran, Kirsten L. Todd, Emma V. Petley, Sherly Mardiana, Christina Mølck, Gregory D. Stewart, Benjamin Solomon, Ian A. Parish, Paul J. Neeson, Simon J. Harrison, Lev M. Kats, Imran G. House, Phillip K. Darcy, Paul A. Beavis
Abstract
Abstract Adenosine is an immunosuppressive factor that limits anti-tumor immunity through the suppression of multiple immune subsets including T cells via activation of the adenosine A 2A receptor (A 2A R). Using both murine and human chimeric antigen receptor (CAR) T cells, here we show that targeting A 2A R with a clinically relevant CRISPR/Cas9 strategy significantly enhances their in vivo efficacy, leading to improved survival of mice. Effects evoked by CRISPR/Cas9 mediated gene deletion of A 2A R are superior to shRNA mediated knockdown or pharmacological blockade of A 2A R. Mechanistically, human A 2A R-edited CAR T cells are significantly resistant to adenosine-mediated transcriptional changes, resulting in enhanced production of cytokines including IFNγ and TNF, and increased expression of JAK-STAT signaling pathway associated genes. A 2A R deficient CAR T cells are well tolerated and do not induce overt pathologies in mice, supporting the use of CRISPR/Cas9 to target A 2A R for the improvement of CAR T cell function in the clinic.