Mechanistic study of Wee1 kinase inhibition with AZD1775 exposes drug targetable vulnerabilities in acute B-lymphoblastic leukemia [scRNA-seq PDX MLL-7]
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 Overall design: scRNA-seq samples were generated from NSG mice transplanted with PDX MLL-7 treated for 24 hours (2 doses) with 120 mg/kg AZD1775 (HY-10993). 4 hours after the last dose mice were sacrificed and bone marrow cells were viably frozen. Before the library preparation, freshly thaw alive hCD45pos hCD19pos mCD45neg bone marrow cells were sorted and proceeded to 10x library protocol. For combination treatments, animals were treated with 6 days of AZD1775 (MedChemExpress) at 120mg/kg alone or with Dasatinib for 3 days (MedChemExpress) at 50 mg/kg. Vehicle animals received 6 days of 5% DMSO, 40% PEG 300, 5% Tween 80 and 50% NaCl solution (all from Sigma-Aldrich). Animals were sacrificed by cervical dislocation 20 hours after the last treatment. Using fresh bone marrow cells from day 6 sequentially treated samples, the cells were FACS sorted like the shorter time point and then proceeded to the 10x library protocol. ***RAW data not provided due to patient privacy concerns***