Fig. 7: Combination therapies target RAS-dependent mTORC1 signaling.

a Workflow for high-throughput combinatorial drug screens of everolimus treatment with the MIPE 5.0 small molecule library. b Heat-map view of the everolimus drug-interaction landscape in SKMM1 and RPMI 8226. Drug synergy is ranked by average aggregate excess highest single agent (HSA) values for SKMM1 and RPMI 8226. Drugs targeting MEK and ERK are indicated on the right. c MEK inhibitors Enrichment Plot from the Drug Set Enrichment Analysis (DSEA) of the Everolimus vs MIPE5.0 screen. The average Excess HSA (SKMM1 and RPMI 8226) was used to pre-rank combinatorial outcomes before running DSEA. P = 0.000; FDR = 0.000. d Response matrix and Excess HSA matrix for the everolimus vs. trametinib (MEKi) combination are shown (SKMM1). e ERK inhibitors Enrichment Plot from the DSEA of the everolimus vs. MIPE 5.0 screen. P = 0.000; FDR = 0.027. f Response matrix and Excess HSA matrix for the everolimus vs. ulixertinib (ERKi) combination are shown (SKMM1). g Cell viability assays for cells treated with DMSO (blue) and 25, 50 or 100 nM everolimus (purples) with the listed doses of trametinib (x-axis) for indicated MM or adenocarcinoma cell lines. Error bars represent SEM of the 3 technical replicates. Representative data; n = 3. h Tumor volume for SKMM1 xenografts treated with vehicle (black), 1 mg/kg trametinib (blue), 1 mg/kg everolimus (green) or the combination (pink). Representative data; n = 3. Error bars represent SEM of tumor size for the mice within each group. i Kaplan–Meier plot indicating survival for SKMM1 xenograft mice. Source data are provided as a Source Data file.