Fig. 2: NRF2 activation promotes CD8+ T cell dysfunction in tumors.
From: The prostacyclin receptor PTGIR is a NRF2-dependent regulator of CD8+ T cell exhaustion
a, Weighted nearest neighbor UMAP (wnn_UMAP) of cell-specific transcripts for the five major activated (CD44+) CD8+ T cell clusters in B16 tumors (14 d.p.t.i.): Tpex, Ttex, TexKLR and TexISG cells and proliferating (prolif.) cells. Gene expression density plots for Cdk1, Pdcd1, Lag3, Tox and Havcr2 are shown. b, GSEA embedding of NRF2 transcriptional target signature (M2870) in the CD8+ TIL UMAP from a. c, Violin plot of NRF2 (M2870) pathway activity score for each CD8+ T cell cluster defined in a. d, B16-OVA melanoma tumor growth in mice after adoptive transfer (A.T.) of WT or Keap1−/− OT-I CD8+ T cells at 8 d.p.t.i. (n = 8–11 mice per group). e, PD-1 versus TIM-3 expression on WT and Keap1−/− OT-I CD8+ TILs isolated from B16 tumors at 15 d.p.t.i. (n = 5 mice per group). f,g, Histogram of PD-1 (f) and TIM-3 (g) expression on WT or Keap1−/− OT-I CD8 TILs isolated from B16 tumors as in e, with gMFI shown in the inset. the bar graphs quantify the gMFI of PD-1 and TIM-3 expression (n = 5 mice per group). h, Representative flow cytometry plot of IFNγ versus granzyme B (GZMB) expression for PD-1Hi WT and Keap1−/− TILs isolated as in e and re-stimulated with phorbol myristate acetate (PMA) plus ionomycin in vitro. i–k, Representative histograms and bar graph of gMFI (n = 5 mice per group) for IFNγ (i), GZMB (j) and TBET (k) expression by PD-1Hi WT or Keap1−/− OT-I CD8+ TILs isolated from B16 tumors as in e. l, B16-OVA melanoma tumor growth in mice after adoptive transfer of WT sgScr or WT sgNfe2l2 OT-I CD8+ T cells at 12 d.p.t.i. (n = 12–17 mice per group). m,n, Functional analysis of PD-1+ WT sgScr or WT sgNfe2l2 OT-I CD8+ TILs isolated from B16 tumors at 18 d.p.t.i. m, Representative flow cytometry plots of IFNγ versus GZMB expression for PD-1+ TILs. The bar graph quantifies the percentage of IFNγ+ GZMB+ donor TILs within tumors (n = 4 mice per group). n, Bar graphs quantifying gMFI of IFNγ, GZMB, TNF and TOX levels in PD-1+ WT sgScr versus WT sgNfe2l2 TILs (n = 4 mice per group). Data are indicated as mean ± s.e.m. (d–n). For tumor plots (d and l), statistical significance was calculated via two-way ANOVA. For all other bar plots (e–k, m and n) statistical significance was calculated via two-tailed, unpaired Student’s t-test. NS (nonsignificant), P ≥ 0.05; *P = 0.05; **P = 0.01; ***P = 0.001; ****P = 0.0001.
