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, Deborah Meyran, Kirsten L. Todd, Emma V. Petley, Sherly Mardiana, Christina Mølck, Gregory D. Stewart, Benjamin J. Solomon, Ian A. Parish, Paul J. Neeson, Simon J. Harrison, Lev M. Kats, Imran G. House, Phillip K. Darcy, Paul A. Beavis

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53 Citations (Scopus)

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 A2A receptor (A2AR). Using both murine and human chimeric antigen receptor (CAR) T cells, here we show that targeting A2AR 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 A2AR are superior to shRNA mediated knockdown or pharmacological blockade of A2AR. Mechanistically, human A2AR-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. A2AR deficient CAR T cells are well tolerated and do not induce overt pathologies in mice, supporting the use of CRISPR/Cas9 to target A2AR for the improvement of CAR T cell function in the clinic.

Original languageEnglish
Article number3236
Number of pages18
JournalNature Communications
Volume12
Issue number1
DOIs
Publication statusPublished - Dec 2021

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