1. CDs with KRASG12C cooperatively sustain downstream signaling outputs to promote survival and proliferation.
CDs with KRASG12C cooperatively sustain downstream signaling outputs to promote survival and proliferation. (A) Immunoblots of H358 and MIA PaCa-2 cells treated with ARS-1323-alkyne (10 μM) simultaneously with DMSO, EGF (100 ng/ml), FGF2 (100 ng/ml), erlotinib (10 μM, EGFRi), AZD4547 (10 μM, FGFRi), bemcentinib (10 μM, AXLi), SHP099 (10 μM, SHP2i), buparlisib (10 μM, PI3Ki), or palbociclib (10 μM, CDK4/6i) for 2 hours. Lysates were subjected to copper-catalyzed click reaction with TAMRA-N3. Cells were grown in 3D spheroid culture. Immunoblots represent two biological replicates. (B) Combination indices (CIs) derived from drug synergism in three KRASG12C-mutant cell lines assessed in 3D spheroid culture. Cell viability was determined after 5-day treatment with ARS-1620, second compounds, or the combination in a 1:1 ratio (dilution series from 1.5 nM to 10 μM), and CI values were calculated using CompuSyn 1.0 from three biological replicates. CI < 0.75 indicates synergism with ARS-1620 (red), CI = 0.75 to 1.25 indicates additivity, and CI > 1.25 indicates antagonism. (C) Clonogenic assays of three KRASG12C-mutant cell lines cultured with indicated compounds or combinations thereof in 2D adherent culture. H358 (300 nM), MIA PaCa-2 (1 μM), and H23 (1 μM) cells were treated using ARS-1620 and second compounds at a 1:1 concentration. Data are representative of three biological replicates.
Kevin Lou et al., Sci. Signal. 2019;12:eaaw9450
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