Fig. 7: MIF is an actionable and selective therapeutic target via Hsp90 inhibition in CRC-derived organoids.
A, C Therapeutic selectivity of Hsp90 inhibitors. Representative images show colonic organoids derived from matched pairs of pooled Mif+/+ tumors or their adjacent epithelium (‘normal’). The indicated organoids were treated with DMSO (‘con’), 17AAG (A), Ganetespib (‘Ganet’), or Onalespib (C) at the indicated concentrations for 21 h. Scale bars, 200 µm. (Right) Quantifications based on organoid morphology. At least 6 images from ≥5 gel domes (×4 magnification each) per group were counted. For each group, number of dead organoids was calculated by the number of total organoids. The data are presented as the means ± SD from different images. B, D Hsp90 inhibitor treatment as described in (A, C) of matched pairs. Immunoblot analysis was performed to evaluate Mif degradation. ‘Mif high’ and ‘Mif low’ indicate exposure times during signal acquisition. Actin, loading control. PC, positive control. Mif/Actin, Mif expression ratios compared to Actin. E Representative images and quantification of organoids from patients with resectable CRC treated with 80 nM Ganetespib for 21 h. Scale bars, 200 µm. Quantification was performed as described in (A). Five images (×10 magnification) per condition were quantified. Means ± SD from different images. A, C, E Student’s t test was performed for comparison of indicated groups; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.
