Sek and Chen et al. demonstrated that overexpression of the adenosine receptor A1R in CAR T cells reduced cAMP levels, and enhanced baseline and antigen-induced cytokine production and effector differentiation. However, these cells exhibited early terminal differentiation in vivo, leading to reduced persistence and loss of the stem-like memory fraction. A CRISPR KI approach was used to restrict A1R expression to the tumor site via the NR4A2 promoter, which enhanced the efficacy of the CAR T cells against solid tumors. Gene network analysis identified the transcription factor IRF8 to play a major role downstream of A1R signaling.
Contributed by Morgan Janes
ABSTRACT: The efficacy of Chimeric Antigen Receptor T cells against solid tumors is limited by immunosuppressive factors in the tumor microenvironment including adenosine, which suppresses Chimeric Antigen Receptor T cells through activation of the A2A receptor. To overcome this, Chimeric Antigen Receptor T cells are engineered to express A1 receptor, a receptor that signals inversely to A2A receptor. Using murine and human Chimeric Antigen Receptor T cells, constitutive A1 receptor overexpression significantly enhances Chimeric Antigen Receptor T cell effector function albeit at the expense of Chimeric Antigen Receptor T cell persistence. Through a CRISPR/Cas9 homology directed repair "knock-in" approach we demonstrate that Chimeric Antigen Receptor T cells engineered to express A1 receptor in a tumor-localized manner, enhances anti-tumor therapeutic efficacy. This is dependent on the transcription factor IRF8 and is transcriptionally unique when compared to A2A receptor deletion. This data provides a novel approach for enhancing Chimeric Antigen Receptor T cell efficacy in solid tumors and provides proof of principle for site-directed expression of factors that promote effector T cell differentiation.
Author Info: (1) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. kevin.sek@petermac.org. Sir Peter MacCallum Department of Oncology, The University of Melbo
Author Info: (1) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. kevin.sek@petermac.org. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. kevin.sek@petermac.org. (2) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (3) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (4) University of Western Australia, Perth, WA, Australia. Telethon Kids Institute, Perth, WA, Australia. (5) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (6) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (7) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (8) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (9) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (10) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (11) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (12) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (13) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (14) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (15) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (16) Frazer Institute, Faculty of Medicine, The University of Queensland, Woolloongabba, QLD, Australia. (17) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (18) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (19) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (20) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (21) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (22) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (23) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (24) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (25) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (26) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (27) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (28) Asthma and Airway Disease Research Center, The University of Arizona, Tucson, AZ, USA. (29) University of Western Australia, Perth, WA, Australia. Telethon Kids Institute, Perth, WA, Australia. (30) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (31) Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (32) Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC, Australia. (33) Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (34) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. (35) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. phil.darcy@petermac.org. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. phil.darcy@petermac.org. (36) Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. paul.beavis@petermac.org. Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. paul.beavis@petermac.org.
