Project name: PSMG2-controlled proteasome-autophagy balance mediates the tolerance for MEK targeted therapy in TNBC

Description: We performed an unbiased genome-wide CRISPR/Cas9 knockout screening to identify synergistic drug targets in TNBC cells with MEK inhibitor. PSMG2 was found as a critical synthetically lethal gene with MEK inhibition. Mechanism investigation unraveled that PSMG2 knockdown inhibited proteasome function, which in turn activated cell autophagy. Autophagy, unlike its function in chemoresistance, inhibited PI3K/AKT pathway by specific degradation of PDPK1. Co-targeting MAPK pathway and proteasome pathway synergistically suppressed TNBC cells proliferation in vitro and in vivo.Overall design: BT549 cells pre-expressing Cas9 were infected with a lentiviral sgRNA library (GeCKO V2) consisting of six single guide ribonucleic acids (sgRNAs) per gene targeting 19050 human genes. The titer of virus added to cells was carefully manipulated to get a multiplicity of infection (MOI) as low as 0.3, ensuring only one sgRNA per cell. Cells were subsequently applied to a puromycin selection to remove the cells without infection. The cells were then split into three parts. One group was directly applied to sgRNA sequencing and labeled as DMSO-Day0. The other two groups, which were labeled as DMSO-Day7 and AZD-Day7, were treated with DMSO and AZD6244 (MEK inhibitor), respectively. After 7 days, the cells were collected and subjected to next-generation sequencing (NGS) of the sgRNA sequences

Data type: Other

Sample scope: Multiisolate

Relevance: Medical

Last updated: 2022-06-23