Figure 1 from A Kinome-Wide Synthetic Lethal CRISPR/Cas9 Screen Reveals That mTOR Inhibition Prevents Adaptive Resistance to CDK4/CDK6 Blockade in HNSCC

CRISPR screening identified the cell-cycle pathway as the synthetic lethal pathway for mTORis in HNSCC. A, Scheme for CRISPR screening. Cal27-Cas9 cells were infected with the human kinome CRIPSR-knockout library, targeting 763 genes consisting of four sgRNAs for each gene, and subjected to vehicle treatment or INK128 treatment. At population doubling 20, genomic DNA was extracted from cells, and PCR and NGS were performed. B, Volcano plot of all dropout sgRNA hits. C, Extraction sequence of sensitizing sgRNA hits. The sensitizing hits were extracted with P value D, Selected 109 hits plot. The hits included genes related to PI3K/mTOR and cell-cycle pathways. E, KEGG pathway analysis for sensitizing hits. KEGG pathway analysis was applied for significant 109 hits. The top 15 pathways are represented, and the color intensity of the bar represents the combined score. F,CCND1 mRNA expression and OS in TCGA-HNSCC patients. Patients with high (z-score > 0.5, n = 124) and low (Z-score n = 73) expression of each mRNA were compared using the log-rank test. G, CCND1 RPPA expression and OS in TCGA-HNSCC patients. Patients with high (z-score > 0.5, n = 120) and low (z-score n = 120) expression of each mRNA were compared using the log-rank test. (Created with BioRender.com.)