Time-course SILAC proteomics to determine the response to WEE1i in RPE-1 WT vs EIF2A-KO RPE-1 cells

AZD1775 is an ATP-competitive inhibitor of WEE1 protein kinase. Inhibition of WEE1 causes hyperactivation of CDK1 and CDK2, which leads to premature mitosis and DNA replication stress. AZD1775 has been evaluated in patients with relapsed high grade serous ovarian cancer (HGSOC) and has shown promising results. However, not all HGSOC tumors responded to AZD1775 treatment, which calls for further investigation into the mechanisms of AZD1775 sensitivity. Based on a viral mutagenesis genome-scale screen in HAP1 cells to identify gene mutations that confer sensitivity to AZD1775 treatment, we showed that inactivation of the alternative translation initiation factor EIF2A confers sensitivity to AZD1775 in several cell line models, including RPE1 cells. Here we measured the differential response of RPE-1 WT or EIF2A-KO cells to WEE1i, at 4 different incubation times.