Fig. 2: Combination PKMYT1i-ATRi is synergistic in OVCA and EMCA cells depending on CCNE1 level.
From: Targeting CCNE1 amplified ovarian and endometrial cancers by combined inhibition of PKMYT1 and ATR
A ZIP synergy scores at various dose combinations of RP-6306 and RP-3500 in FT282-hTERT p53R175H parental (WT) and CCNE1-overexpressing (CCNE1-O/E) cells. Score ≥ 10 (red color) represents synergy, ≤ − 10 (green) represents antagonism. Values were obtained by analyzing mean data from 3 independent biological replicates with SynergyFinder. B Growth inhibition relative to DMSO control of parental and CCNE1-overexpressing cells after treatment with the indicated dose of RP-6306, RP-3500, or the combination of both. n = 3; Mean + SD. C, D Cell viability detection of SNU685 cells in response to RP-6306 monotherapy (C) and RP-3500 monotherapy (D). n = 3; Mean ± SD. Highlighted dose showing the statistical difference in CCNE1-induced SNU685 cells with or without CCNE1 induction. E, F Cell viability analysis of the indicated SNU685 CCNE1 inducible cells ± doxycycline lines after treatment at the indicated doses. Doxycycline: 1 µg/ml. Monotherapy for PKMYT1i, RP6306, is highlighted in blue, for ATRi, RP3500, is highlighted in red, Combinations are highlighted in purple for PKMYT1i-ATRi. Assays were normalized by doubling time such that cells doubled at least twice. n = 3; Mean ± SD. Growth inhibition relative to DMSO control of parental and CCNE1-overexpressing cells after treatment with indicated doses in pink (middle). The coefficient of drug interaction (CDI) relative to the fraction affected (Fa) plot of the indicated cell lines is indicated to the right of each bar graph. CDI < 1 synergy with CDI < 0.7 significant synergy, CDI = 1 additive, CDI > 1 antagonistic. The red dot corresponds to doses highlighted in pink. G Measurement of drug combinations in WO-20 CCNE1inducible cells with or without Cyclin E1 induction. n = 4; Mean + SD. H, I Colony formation analysis (Upper panels) and quantification (Lower panels) of WO-20 CCNE1inducible cells (H) and SNU685 CCNE1inducible (I) in response to RP-6306 (31.3 nM), RP-3500 (6.25 nM) or combination for 10 days. Significance determined by two-way ANOVA followed by Tukey’s multiple comparisons test for (B, E, G) and two-way ANOVA followed by Tukey’s multiple comparisons for (C, D).
