Extended Data Fig. 10: CDKN1B ablation improves the expansion of T cells engineered with a cilta-cel-like BCMA-targeting CAR.
From: In vivo CRISPR screens identify modifiers of CAR T cell function in myeloma
a. MM.1S tumor burden as assessed by bioluminescence imaging (BLI) of mice treated with intergenic control KO or CDKN1B KO BCMA cilta-cel-like CAR-T cells. NSG mice were injected intravenously with 1E6 MM.1S followed by transfer of CAR-T cells (ND116) 21 days later. n = 5 mice in tumor only and control KO groups; n = 6 mice in CDKN1B KO group. b. Survival curve, n = 5 mice in tumor only and control sgRNA groups; n = 6 mice in CDKN1B sgRNA 1 group per group. c-d. Percentage (c) and numbers (d) (mean +/– SEM) of intergenic control KO (n = 5 mice) or CDKN1B KO (n = 6 mice) CAR-T cells in the peripheral blood at the indicated time post CAR-T cell infusion. Two-way ANOVA with Tukey’s multiple comparison test. e. MM.1S tumor burden (mean +/– SEM, n = 6 mice in tumor only and CDKN1B sgRNA 1 groups; n = 5 mice in intergenic control KO group) as assessed by BLI in mice treated with intergenic control KO or CDKN1B KO BCMA cilta-cel-like CAR-T cells (ND202). f-g. Percentage (f) and numbers (g) (mean +/– SEM, n = 6 mice in CDKN1B sgRNA 1 group; n = 5 mice in control sgRNA group) of intergenic control KO or CDKN1B KO CAR-T cells the peripheral blood at the indicated time post CAR-T cell infusion. Two-way ANOVA with Tukey’s multiple comparison test. h. CD8/CD4 CAR-T cells ratio (mean +/– SEM, n = 6 mice in CDKN1B KO group; n = 5 mice in intergenic control KO group) over time in the peripheral blood of the mice. i-j. Phenotype of intergenic control KO or CDKN1B KO (n = 6 mice) CD4+ and CD8+ CAR-T cells at EOP (i) or in the mice (j, mean +/– SEM, n = 6 mice in CDKN1B sgRNA 1 group; n = 5 mice in control sgRNA group) at the indicated time post CAR-T cell infusion. T memory stem cells, (TSCM, CD45RA + CCR7 + CD95+), central memory (CM, CD45RA-CCR7+), effector memory (EM, CD45RA-CCR7-) and terminally differentiated effector memory (TEMRA, CD45RA + CCR7-) T cells. Statistical significance measured by two-sided Student’s t-test. Day21: CD8 + CAR-T: TSCM *p = 0.0138; Day28: CD4 + CAR-T: TEMRA *p = 0.0328. k. Percentage (mean +/– SEM) of PD1, TIM3 and CD39 expression in intergenic control KO (n = 5 mice) or CDKN1B KO (n = 6 mice) CD4+ and CD8+ CAR-T cells in the peripheral blood of the mice at the indicated time post CAR-T cell transfer. Two-way ANOVA with Tukey’s multiple comparison test.
