Additional file 1 of High-throughput drug screening identifies EGFR/MAPK pathway targeting sensitivities in organoid models of ovarian carcinosarcoma

Supplementary Material 1: Supplementary Figure S1: Drug screening of a mouse OCS cell line identifies sensitivity to independent cisplatin- and eribulin-based combinations. a) Dose response curves for eribulin and cisplatin in the mouse OCS cell line (left), validation of EC20 doses (right), and surviving fractions for chosen EC20 concentrations (bottom); b) Waterfall plot of cisplatin-based combination drug effects in the initial screen. DEffect values are calculated by subtracting the effect of the compound alone from the effect of the compound in combination with cisplatin. Most effective cisplatin-based combinations are indicated on the right as well as compounds selected for the validation screens (green); c) Waterfall plot of eribulin-based combination drug effects in the validation screen. DEffect values are calculated by subtracting the effect of the compound alone from the effect of the compound in combination with eribulin. Most effective eribulin-based combinations are indicated on the right as well as compounds selected for the validation screens (green). SF, survival fraction. Supplementary Figure S2: Validation of EMT-related single agent hits from the drug screen in cell lines. a) Dose response curves for each compound in four cell lines (two OCS and two HGSOC); b) EC50 values for each of the EMT-related single agents tested in cell lines; c) Heatmap of EC50 values for each EMT-related single agent in each cell line; d) Average EC50 values for the HGSOC cell lines combined compared to OCS cell lines combined. Supplementary Figure S3: Validation of N-MYC-related single agent hits from the drug screen in cell lines. a) Dose response curves for each compound in four OCS cell lines; b) EC50 values for each of the N-MYC-related single agents tested in cell lines; c) Heatmap of EC50 values for each N-MYC-related single agent in each cell line. Supplementary Figure S4: Validation of all single agent hits from the drug screen in organoids. a) Dose response curves for EMT-related compounds in seven organoid models (two HGSOC and five OCS); b) Dose response curves for N-MYC-related compounds in nine organoid models (three HGSOC and six OCS). Supplementary Figure S5: Western blot showing increased expression of N-MYC in OCS. Expression levels of N-MYC were validated via Western blot. Supplementary Figure S6: Validation of cisplatin- and eribulin-based combinations in OCS cell lines. Cell lines were treated with 6-point dose range of cisplatin or eribulin in combination with 6-point dose range of selected compounds and HSA and CSS values were calculated; a) HSA synergy heatmap for cisplatin-based combinations in OCS cell lines; b) CSS values for cisplatin-based combinations; c) SS plot for cisplatin-based combinations in OCS cell lines; d) HSA synergy heatmap for eribulin-based combinations in OCS and HGSOC cell lines; e) CSS values for eribulin-based combinations; f) SS plot for eribulin-based combinations in OCS and HGSOC cell lines. HSA, highest single agent; CSS, combination sensitivity score; SS, synergy score. Supplementary Figure S7: Imaging and tracking of drug treated organoids. The growth and changes to architecture of the organoids were tracked over time by imaging once a day for 7 days after drugs were added. In this example, a matrix of cisplatin and adavosertib were added to #233 organoids and their growth was tracked over time. An example of the change in organoid appearance following drugging can be seen in the panels on the right. Supplementary Figure S8: Triple combination of eribulin with BGB-283/mirdametinib increases drug synergy in OCS cell lines. a) HSA synergy plots for PH142 cells treated with a BGB-283/mirdametinib drug matrix alone or (b) with a single dose of eribulin; c) HSA synergy plot for PH003 cells treated with a BGB-283/mirdametinib drug matrix alone or (d) with a single dose of eribulin; e) Bar graphs showing CSS values for both PH142 and PH003 cells treated with double or triple combination therapy; f) SS plot for PH142 and PH003 cells treated with double or triple combination therapy. HSA, highest single agent; CSS, combination sensitivity score; SS, synergy score.