Fig. 5.

Change in effect of therapy when adding a second inhibitor predicted by the computational model, and successful MEK and COX2 inhibition combination therapy in vivo. (A and B) Rows and columns of heatmaps are colored by pathway to which nodes which are treated belong; the pathway categories are laid out in Dataset S7. Mean change in proliferation (A) or apoptosis (B) across both clones when adding a second inhibitor to an inhibitor already shown to be effective in monotherapy. In the heatmap, the y axis shows first inhibitor; x axis, the inhibitor that is added in combination. The gray boxes are combinations that are nonsensical (2 different inhibitions of the same node) or cause apoptosis above 3 in the healthy cells. Inhibition of PHD2 or VHL cause high apoptosis with every partner other than each other and are removed as this otherwise prevents clustering of the heatmaps. (C) Caliperimetric measurement of tumor growth of mixed WM⁺T/WM⁻T tumors during 3 d of treatment with tamoxifen (100 μg/mouse, twice daily) followed by 4-d treatment with tamoxifen and drug combinations (celecoxib, 20 mg/kg/d; PD0325901, 10 mg/kg/d). Measurements are normalized to the beginning of the drug treatment course as tumors at this stage were at different sizes (n = 4 to 5; error bars represent SD). (D) Representative picture of the gross morphology of mixed WM⁺T/WM⁻T tumors after the treatment described above. (E) Quantification of cell death in mixed WM⁺T/WM⁻T tumors as percentage of CC3-positive pixels in the individual clones (n: control, PD0325901 = 4; celecoxib, celecoxib plus PD0325901 = 5). (F) Quantification of proliferation in mixed WM⁺T/WM⁻T tumors by automated counting of IDU-positive nuclei over total nuclei in the individual clones. (n: control, PD0325901 = 4; celecoxib, celecoxib plus PD0325901 = 5). For box-and-whisker plots, the error bars represent min to max values, the box represents the interquartile range, and the horizontal line represents the median. P values are based on Student’s t test: *P < 0.05.