Project description:we demonstrate that the WEE1 inhibitor AZD1775 triggers endoplasmic reticulum (ER) stress and activates the PERK and IRE1α branches of the unfolded protein response (UPR) in TP53 mutant HGSOC cells. Upon AZD1775 treatment, PERK facilitates apoptotic signaling in these cells via activating CHOP, whereas IRE1α-induced spliced XBP1 (XBP1s) confers survival in response to WEE1 inhibition. Our data uncover an important dual role of UPR in TP53 mutant HGSOC cells in response to AZD1775, where additional inhibition of IRE1α-XBP1s signaling may offer synergistic efficacy.
Project description:The aim of this study was to characterize phosphoproteome in response to AZD1775 in patient derived xenografts (PDX) lines and clinical specimens in glioblastoma. Tyrosine and motif specific serine and threonine phosphorylation were measured using quantitative LC-MS/MS based on TMT labeling.
Project description: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.
Project description:CRISPR-Cas9 genome-wide screens were performed in retinal pigment epithelial cells (RPE1) with either wild-type TP53 gene, or a TP53-null background. Results show wild-type TP53 has minimal impact on the efficiency of CRISPR dropout screens.
Project description:This study characterized the effect of WEE1 kinase inhibition using AZD1775 treatment on single-cell accessible chromatin and gene expression profile in the acute lymphoblastic leukemia cell line RS4;11 that represents the KMT2A-rearranged subtype.
Project description:This study characterized the effect of WEE1 kinase inhibition using AZD1775 treatment on single-cell gene expression profile using the 10x Genomics protocol in the acute lymphoblastic leukemia cells
Project description:This study characterized the effect of WEE1 kinase inhibition using AZD1775 treatment on single-cell gene expression profile using the 10x Genomics protocol in the acute lymphoblastic leukemia cell lines RS4;11 (KMT2A-rearranged subtype) and Nalm6 (other subtype).