Poster Presentation Asia-Pacific Vaccine and Immunotherapy Congress 2026

Tumor site directed A1R expression enhances CAR T cell function and improves efficacy against solid tumors (#140)

Kevin sek 1 , Amanda Chen 1 , Thomas Cole 1 , Paul Beavis 1 , Phillip Darcy 1
  1. Peter MacCallum Cancer Centre, The University of Melbourne, Darcy Lab, Rowville, VIC, Australia

The efficacy of Chimeric Antigen Receptor (CAR) T cells against solid tumours is limited by immunosuppressive factors including adenosine, which suppresses CAR T cells through activation of the A2A receptor (A2AR). We have previously demonstrated that CRISPR/Cas9 deletion of the A2AR significantly enhances the in vivo efficacy of both mouse and human CAR T cells [1]. Alternatively, CAR T cells engineered to express A1R, a receptor that signals inversely to A2AR, significantly enhanced the function of CAR-T cells. Constitutive A1R expression however promoted T cell differentiation and exhaustion leading to reduced CAR T cell persistence in vivo. To overcome this limitation, CRISPR/Cas9 homology directed repair was used to “knock-in” A1R under the control of the NR4A2 promoter, leading to precise expression of A1R only at the tumour site but not other tissues [2]. This led to effector differentiation and enhanced anti-tumour efficacy in-vivo, concomitant with improved long-term persistence. We further identified through CRISPR deletion, the transcription factor IRF8 as a major hub factor downstream of A1R signalling. Critically, IRF8 has not been previously implicated in T cell function and effector differentiation. These data provide a novel approach for rewiring T cell differentiation factors to enhance CAR T cell efficacy in solid tumours and could be utilized to deliver other ‘precision-guided’ payloads specifically to the tumour site [3].

  1. Giuffrida, L., et al., CRISPR/Cas9 mediated deletion of the adenosine A2A receptor enhances CAR T cell efficacy. Nat Commun, 2021. 12(1): p. 3236.
  2. Sek, K., et al., Tumor site directed A1R expression enhances CAR T cell function and improves efficacy against solid tumors. Nat Commun, 2025.
  3. Chen, A.X.Y., et al., Rewiring endogenous genes in CAR T cells for tumour-restricted payload delivery. Nature, 2025.