Fig. 4: RASA2 ablation improves in vivo tumour control by engineered T cells in multiple preclinical models.
From: RASA2 ablation in T cells boosts antigen sensitivity and long-term function
a,b, NY-ESO-1+ A375 melanoma cells were engrafted into NSG mice via flank injection and NY-ESO-1-specific TCR T cells were injected via the tail vein. a, Experimental timeline. b, Tumour growth was monitored with calliper measurements (n = 6 mice per group; mean ± s.e.m.; two-sided unpaired Student’s t-test). c,d, NY-ESO-1+ Nalm6 leukaemia cells were injected into NSG mice followed by NY-ESO-1-specific TCR T cells. c, Experimental timeline. BLI, bioluminescence live imaging. d, Tumour growth was monitored using luciferase-based bioluminescence live imaging (n = 5 mice for RASA2-KO T cells, n = 4 for control T cells; mean ± s.e.m.; two-sided unpaired Student’s t-test). e,f, Nalm6 cells were injected into NSG mice followed by CD19-specific CAR T cells. e, Experimental timeline. f, Tumour growth was monitored by bioluminescence imaging (n = 7 mice per group; mean ± s.e.m.; two-sided unpaired Student’s t-test). g, Bioluminescence imaging of the cohort in f, dorsal view. h, Survival of the cohort shown in f. i, Cell counts by flow cytometry in bone marrow of Nalm6-engrafted NSG mice (day 7: n = 5 for control, n = 6 for RASA2 KO; day 16: n = 6 per group; mean ± s.e.m.; two-sided Wilcoxon test). j, Mean fluorescence intensity (normalized to control) of inhibitory markers on cells from cohort in i (mean ± s.e.m.; two-sided Wilcoxon test). k, Percentage of mixed CAR T cell population (originally injected into mice, mixed 50:50 (control:RASA2-KO CAR T cells)), isolated from bone marrow days 7 and 16 after infusion into Nalm6-bearing mice (n = 6 mice per group; two-sided Wilcoxon test). l–o, NSG mice were injected intraperitoneally with LM7-ffLuc tumour cells on day 0, then received a single intraperitoneal injection of control or RASA2-KO EphA2-CAR T-cells. l, Experimental timeline. m, Quantitative bioluminescence imaging (mean ± s.e.m.; n = 10 for control, n = 14 for RASA2 KO; two-sided paired Student’s t-test). n, Representative bioluminescence for each group. o, Survival curve for the cohort in m. Survival P-values by log-rank test. Statistical tests as indicated. *P < 0.05, **P < 0.01, ****P < 0.0001.
