Fig. 5: The organoid model detects donor-dependent differences in TCB-triggered toxicity.
a, Representative single tiles of merged brightfield and IF images of the 14-organoid donor lines co-cultured with PBMCs treated with EpCAM, CEA(hi), CEA(lo) and NT TCB (0.5 µg ml−1) at 48 h post TCB administration. ×5 magnification; scale bar per tile, 1 mm. Organoid donor number is random, following no particular order. b, Heat map of quantified caspase-3/7 fluorescence signal in dozens of segmented organoids per well (n = 3) for each TCB treatment across time. Mean fluorescence signal for each TCB condition normalized to the mean a.f.u. detected in the NT TCB control at each timepoint. The 14 different patient-derived organoid lines are displayed on the x axis and ordered by extent of apoptosis experienced. c, Representative IHC of CEA (top) and EpCAM (bottom) expression of the 14 embedded organoid lines at ×40 magnification. Organoid donors ordered by expression levels. Scale bar, 50 µm. d, Quantified target expression levels of cross sections in c for CEA (top) and EpCAM (bottom) indicated as positive area for each individual organoid. Donors are ordered by increasing target expression. e, Data in d as boxplots, with whiskers showing all points (minimum to maximum) of the mean target expression across all donors distinguished between the indicated intestinal regions for each organoid line (n = 7). Unpaired t-test (two-tailed) was performed. f, Correlation plots of the target expression between CEA or EpCAM and normalized caspase-3/7 signal of the respective TCB. R2 is provided per plot. g, Data as boxplots, with whiskers showing all points (min. to max.) of the mean of normalized caspase-3/7 across all donors distinguished between the indicated intestinal regions for each organoid line at 48 h post administration (n = 7). Unpaired t-test (two-tailed) was performed. All displayed experiments in this figure were replicated at least three times, yielding similar results.
