Extended Data Fig. 10: BTLA-deficient CAR T cells for solid malignancies.

a, Analysis of overall and disease-free survival of all TCGA cohorts based on BTLA expression normalized to CD3E, performed using GEPIA2 (Methods). Cutoffs are defined by quartiles. b,c, TCGA correlates of TNFRSF14/HVEM and PDL1 tumor expression. b, Transcript expression in log₂(TPM + 1) of HVEM (top) and PDL1 (bottom) across all TCGA cohorts. c. Disease-free survival of all TCGA cohorts, split either by median HVEM expression (left), or median PDL1 expression (right). d, Disease-free survival of TCGA OV cohort, based on BTLA (left) or PDCD1 (right) RNA expression normalized to CD3E (upper and lower quartile cutoffs). e-g, anti-HER2 studies (clone 4D-5, see Methods). e, GFP Integrated Intensity of SK-OV-3 ovarian cancer cells in coculture with anti-HER2 CAR T cells (effector: target = 0.25, n = 5 technical replicates). f, Disease-free survival of TCGA PRAD cohort, based on BTLA expression normalized to CD3E (median cutoff). g, Cell Index (impedence) of PC-3 prostate cancer cells in coculture with anti-HER2 CAR T cells (effector: target = 0.5, n = 3 technical replicates). h, Knockout of HVEM from PC-3 eliminates benefit of BTLA KO in anti-HER2 CAR T cells. Here, 4,000 WT or HVEM-deficient CBG⁺ PC-3 cells were seeded 24 hr prior to the addition of anti-HER2 CAR T cells (n = 3 technical replicates). Luminescence was measured at 72 h of coculture. i, OS of TCGA PAAD cohort, based on BTLA expression normalized to CD3E (median cutoff). j, Low basal HVEM expression on AsPC-1 pancreatic cancer cells. k, Luminescence-based cytotoxicity studies of anti-mesothelin CAR (clone M5, see NCT03323944) T cells (that is, CARTM5) against mesothelin⁺ CBG⁺ HVEM-low AsPC-1 cells. Here, 4,000 AsPC-1 cells were seeded 24 h before the addition of WT or BTLA KO CARTM5 cells at various effector to target ratios (n = 3 technical replicates). Measurements were taken at 48 h and 72 h of coculture and show no benefit of BTLA KO in this setting. l, Cell Index (impedence) of B16-muCD19 cells cocultured with high doses of UTD, WT muCART19, or BTLA KO muCART19 (n = 3 technical replicates, see Methods). m-p, In vivo B16-muCD19 melanoma study (Methods). m,n, Tumor volumes following infusion of WT (n = 8 mice, m) or BTLA KO (n = 9 mice, n) murine CART19 cells. o, Tumor sizes on day 14 following CAR T cell infusion (WT, n = 8; BTLA KO, n = 9) p, Overall survival. For all bar plots, data are presented as mean ± s.d. Statistical significance was determined by one-way ANOVA with post hoc Tukey tests (e,g,l at endpoints), two-way ANOVA (k), two-sided unpaired t-tests (h,o), or long-rank Mantel Cox test (c,d,f,i,p).

Source data