Fig. 3: NR4A2/IL-12 engineered T cells are well tolerated and elicit potent antitumour immunity.
From: Rewiring endogenous genes in CAR T cells for tumour-restricted payload delivery
a,b, OT-I cells engineered to express GFP were assessed after 24-hour stimulation with anti-CD3/CD28 antibodies or ova-expressing tumour cells (a) and in mice bearing AT-3-ova tumours 8 days after transfer (b). Flow cytometry plots (left) and quantification (right) of GFP expression in OT-I cells. c–e, We adoptively transferred 12.5 × 106 mock, NR4A2 knockout or NR4A2/IL-12 OT-I cells into mice with AT-3-ova tumours. Data are tumour growth (c), survival (d) and body weight (e). f, Schematic depicting differences between endogenous and synthetic promoters. g–i, Murine anti-Her2 CAR T cells expressing GFP through synthetic NFAT or endogenous Nr4a2 promoters were assessed after 24-hour stimulation with anti-CD3/CD28, anti-CAR antibodies or hHer2-expressing tumour cells (g) and in E0771-hHer2 tumour-bearing mice 9 days after transfer (h,i). Flow cytometry plots (h) and quantification (g and i) of GFP expression in CD8+ CAR T cells. j,k, 5 × 106 mock, NR4A2 knockout or NR4A2/IL-12 murine anti-hHer2 CAR T cells were adoptively transferred into E0771-hHer2 (j) or MC38-hHer2 (k) tumour-bearing mice. Data are tumour growth (left) and survival (right). a,g, Data are mean ± s.d. of technical triplicates, representative of n = 3 (a) and n = 2 (g) experiments. b–e,h–k, Data are mean ± s.e.m. from n = 6 (NR4A2/GFP) or 7 (PD-1/GFP) mice per group, representative of n = 3 experiments (b); n = 5 mice per group (c–e); n = 3 (tumour) and 4 (non-tumour tissues) mice per group (h,i); n = 4 (NR4A2/IL-12), n = 5 (mock) and n = 6 (non-treated, NR4A2 knockout) mice per group (left), or n = 5 (mock), n = 6 (non-treated, NR4A2 knockout) and n = 7 (NR4A2/IL-12) mice per group (right), representative of n = 2 experiments (j); n = 6 mice per group (k). a–c,g,j–k (Left), two-way ANOVA; d,j–k (right), log-rank Mantel–Cox test; i, one-way ANOVA. NS, not significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Illustrations in f created using BioRender: Chen, A., https://BioRender.com/03b4c93 (2025).
