Fig. 3: Behavioral heterogeneity of TCR and CAR T cell therapies targeting BC PDOs.
From: Uncovering the mode of action of engineered T cells in patient cancer organoids
a,b, Quantification of BC PDO viability using CellTiter-Glo following overnight coculture of PDOs with WT1 T cells (a) or ROR1 CAR T cells (b). a,b, One-way ANOVA followed by Dunnett’s correction: 10T versus 36T, P < 0,0001 (****); 10T versus 169M, P < 0,0001 (****); 10T versus 62T, P < 0,0001 (****) (a); 34T versus 36T, P < 0,0001 (****); 34T versus 169M, P < 0,0001 (****); 34T versus 10T, P < 0.0001 (****) (b). Data corrected for untransduced T cell responses (mean ± s.d.). c,d, 3D multispectral images of BC PDO cultures (yellow) showing killing by WT1 T cells (blue, c) or ROR1 CAR T cells (blue, d) at the indicated time points of imaging. Dead cells depicted in red. Scale bars, 30 µm. e, FACS histogram plots showing ROR1 expression in the indicated breast cancer PDO cultures (blue) compared with unstained control (gray). f,g, Behavioral cluster distribution of WT1 T cells (f) and ROR1 CAR T cells (g) cocultured with the indicated BC PDOs (mean ± s.e.m.). X2-test, P < 0,0001. h,i, Super engager (CL9) cluster size (%) of total for WT1 T cells (h) and ROR1 CAR T cells (i). h, One-way ANOVA with Dunnett’s correction: 10T versus 169M, P = 0,0501; 10T versus 62T, P = 0.0006; 34T versus 36T, P = 0.0018; 34T versus 169M, P < 0.0001; 34T versus 10T, P = 0.0002 (mean ± s.d.). j, Behavioral cluster size difference (%) between TEGs and CAR T cells cocultured with 34T (middle) or 10T (right), or between WT1 T cells and CAR T cells cocultured with 169M PDOs (left) (mean ± s.d.). Welch’s two-sided t-test: 169M: CL1, P = 0.015; CL2, P = 0.041; CL5, P = 0.023; CL6, P = 0.047; CL7, P = 9.94 × 10–4; CL9, P = 0.012. 34 T: CL1, P = 0.004; CL2, P = 0.016; CL3, P = 0.003; CL5, P = 0.012; CL8, P = 0.0004; CL9, P = 0.037. 10T: CL1, P = 0.0014; CL3, P = 0.0045; CL5, P = 0.014; CL6, P = 0.025; CL7, P = 0.001; CL9, P = 1.16 × 10–5. a–e, Representative of n = 3 independent experiments; f–j, n = 3–6 independent experiments; see Supplementary Table 8 for value of n per condition.
