Extended Data Fig. 1. Pharmacological inhibition of FOXO1 impairs expansion, formation of a memory phenotype and antitumour function in CD19.28ζ and CD19.BBζ CAR T cells.
CAR T cells were treated with DMSO or 10 nM or 100 nM of the small molecule AS1842856 (FOXO1i) starting on day 4 post-activation and treated every 2–3 days thereafter. a, Schematic of FOXO1i experimental model. b, CD19.28ζ (left) or CD19.BBζ (right) CAR T cell expansion (n = 2 donors). c, Percent CD8+ in CD19.28ζ (circles) and CD19.BBζ (squares) cells (n = 2 donors for each CAR). d, Apoptosis in CD19.BBζ CAR T cells at day 15 post-activation. Contour plots show 1 representative donor and bar graphs show mean±s.e.m. of n = 3 donors. e, Expression of memory- and exhaustion-associated markers on CD19.28ζ and CD19.BBζ cells. Histograms show 1 representative donor (n = 2 donors). f, Cytokine secretion from CD19.28ζ and CD19.BBζ cells in response to Nalm6 cells. Graphs show mean±s.d. of triplicate wells from 1 representative donor (n = 2 donors). g, Cytotoxicity of CD19.BBζ cells against Nalm6 cells at a 1:1 E:T ratio. Data is normalized to t = 0 and show mean±s.d. of triplicate wells from 1 representative donor (n = 2 donors). Statistics are shown for t = 60 h. Statistical comparisons were performed using paired two-tailed Student’s t-test (c), two-way ANOVA with Šídák’s test (d) and one-way (f) or two-way (g) ANOVA with Dunnett’s test. E:T ratio, effector:target cell ratio. NS, not significant.