Fig. 4. FOXO1OE CAR T cells exhibit enhanced tumour control and sustained effector function in solid tumours.
A total of 5 × 106 mock or tNGFR+ Her2.BBζ CAR T cells expressing tNGFR or FOXO1OE were infused into 143B-bearing mice three days after engraftment. a,b, Tumour measurements over time (a) and on day 25–29 (b). One FOXO1OE mouse has been omitted in b owing to tumour-independent death before day 25. Data were pooled from three donors tested in three independent experiments (n = 11–18 mice per group). c–e, Analysis of day-29 CAR TILs. c, Total CAR TILs (n = 13 mice per group). d, Ratio of CD8+ to CD4+ CAR TILs. One representative donor (n = 10 mice per group). e, CAR TIL IL-2 and IFNγ secretion after ex vivo stimulation with 143B (n = 13 mice per group). Data in c–e were pooled from two donors tested in two independent experiments. f–h, Single-cell RNA-seq on day-29 CAR TILs. Cells were sorted and pooled from n = 5 mice per group from one donor. f, Left, uniform manifold approximation and projection (UMAP) of CAR TILs. Eleven clusters were identified with k-nearest neighbours clustering, and were annotated manually (middle). Right, sample distribution by cluster. Teff, T effector cell; TRM, tissue resident memory T cell. g, Sample distribution within the UMAP. h, Teff, TRM and FOXO1OE-associated transcriptional signatures. Long dashed lines represent the mean and short dashed lines represent the top and bottom quartiles. Data in b–e are mean ± s.e.m. Statistical comparisons were performed using two-tailed Student’s t-test (b–e) and two-sided Wilcoxon rank-sum test (h).