Fig. 1: CBIs cooperate with FMCC to kill pancreatic cancer cells and colorectal tumour organoids.

a Scheme of the screenings for agents which become cytotoxic for cancer cells through FMCC (i.e., 1% FBS and 0.5 g/l glucose). b PK9 PDAC cells were plated in 96-well plates and treated for 96 h w/ or w/o FMCC. During the last 72 h libraries of bioactive or clinically approved compounds were added (at a final 10 µM concentration) before cell viability was detected. A cooperative index ≤0.8 was arbitrarily chosen to select for drugs that synergistically interacted with FMCC. c The cholesterol biosynthesis pathway and some of its inhibitors. d PK9 cells were treated for 96 h w/ or w/o FMCC and for the last 72 h w/ or w/o 10 μM oxiconazole, miconazole or clotrimazole; 1 μM itraconazole; 30 μM simvastatin or terbinafine. Thereafter, cell viability was measured. e Capan-1 cells were treated for 24 h w/ or w/o FMCC, 25 μM clotrimazole, 50 μM terbinafine or their combination before cellular cholesterol content was measured. f Capan-1 cells were treated for 96 h w/ or w/o FMCC and for the last 72 h w/ or w/o 10 μM clotrimazole or 30 μM simvastatin. Where indicated, FMCC were supplemented with 80 µM LDL/HDL cholesterol (at an LDL/HDL ratio of 3:1) to reach a final cholesterol concentration of ~90 µM in the medium. Finally, cell viability was measured. g Patient-derived colorectal cancer organoids (OMCR15-045TK) were treated for 192 h w/ or w/o 15 μM clotrimazole, 20 μM terbinafine, FMCC or their combination. Organoids were imaged daily. Mean organoid area was calculated from ten biological replicates using Image-J. Organoid photographs were obtained on the last day of the experiments. A scale bar is included in the upper left photograph. The drug screen (b) was performed once. Hit compounds (26) were retested twice; 10 of them were confirmed to synergyze with FMCC. In d-f, data points are experimental replicates; data are shown as mean ± SD. In (g) (organoid growth curves), data are presented as mean ± SD of three experimental replicates. p values were calculated by two-tailed Student’s t test. Source data are provided as a Source Data file.