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Journal: Frontiers in Cell and Developmental Biology
Article Title: Inhibiting the RNA helicase DDX3X in Burkitt lymphoma induces oxydative stress and impedes tumor progression in xenografts
doi: 10.3389/fcell.2025.1642006
Figure Lengend Snippet: CRISPR chemogenomic screen identifies modulators of sensitivity and resistance to DDX3X inhibition by RK-33. (A) unranked CRANKS scores for all sgRNA (left) and ranked CRANKS score (right) from highest to lowest, showing sgRNA enrichment (rescues, green) and depleted (sensitizers, red) in RK-33-treated NALM-6 cells. sgRNAs targeting genes known to be involved in oxidative stress response are identified. (B) top 5 enriched (green) and top 5 depleted (pink) sgRNAs are shown, alongside Gene Ontology (GO) analysis for all significantly enriched (top chart) and depleted (bottom chart) sgRNAs. (C) Gene set enrichment analysis (GSEA) for the KEGG Gluthatione Metabolism (left) and KEAP1-NRF2 signaling pathway (right) using RNA-seq data from CA46 and Raji cells treated with RK-33 (as presented in ). (D) Total Reactive Oxygen Species (ROS) in CA46 and Raji cells after 24 h of treatment with DMSO or RK-33, measured by flow cytometry using the FITC channel and reported as mean fluorescence intensity (MFI). Statistical significance was calculated using unpaired t -tests. (E,F) Dose-dependent effects of combinatorial treatments using RK-33 and inhibitors of key hits from (B) , assessed by XTT viability assay after 4 days. CPUY192018 (CPUY) and omaveloxolone (Omav) are KEAP1 inhibitors; inosine and TAK243 are UBA6 inhibitors; and buthionine sulfoximine (BSO) is a GCL inhibitor. (E) Viability curves of cells treated with BSO in combination with RK-33. (F) Calculated CC 50 values for each cell line and treatment combination, shown as box-and-whisker plots. P values were evaluated using a Kruskal–Wallis H test followed by Dunn’s multiple comparisons test comparing each combination treatment to vehicle + RK-33. In all graphs, ** P < 0.01; *** P < 0.001; **** P < 0.0001.
Article Snippet: TAK-243 and omaveloxolone were obtained from Selleck Chemicals, while
Techniques: CRISPR, Inhibition, RNA Sequencing, Flow Cytometry, Fluorescence, Viability Assay, Whisker Assay