Journal: Nature Communications

Article Title: Autocrine interferon poisoning mediates ADAR1-dependent synthetic lethality in BRCA1/2 -mutant cancers

doi: 10.1038/s41467-025-62309-5

Figure Lengend Snippet: A Western blot of SUM149 BRCA1 -Mut and BRCA1 -Rev cells transduced with a doxycycline-inducible ADAR1 -targeting shRNA and exposed to a titration of doxycycline (ng/mL). Data representative of n = 2 biologically-independent experiments. B , C RT-qPCR of IFNB1 ( B ) and CCL5 ( C ) mRNAs in SUM149 BRCA1 -Mut and BRCA1 -Rev cells transfected with a concentration range (nM) of ADAR1 siRNA. IFNB1 and CCL5 mRNAs were analyzed separately relative to GAPDH . Box-and-whiskers show arbitrary units of gene expression normalized to the BRCA1 -mutant siCTRL condition; N = 4 values from individual measurements, representative of n = 3 biologically-independent experiments, two-way ANOVA post hoc Dunnett’s test. P values, ***[ IFNB1 , BRCA1 -Rev, siADAR1 1.25 nM]=0.0002, ****< 0.0001. D Western blot of MEF Brca1 -wildtype (WT) and Brca1 -mutant (Δ11) cells transfected with a concentration range (nM) of Adar1 siRNA. Data representative of n = 2 biologically-independent experiments. E , F RT-qPCR of Ifnb1 ( E ) and Ccl5 ( F ) mRNAs in MEF Brca1 -wildtype (WT) and Brca1 -mutant (Δ11) cells transfected with a concentration range (nM) of Adar1 siRNA. Data presented as in ( B , C ). P values, ****< 0.0001. G , H Clonogenic survival of SUM149 BRCA1 -Mut and BRCA1 -Rev cells subjected to co-transfection with ADAR1 siRNA and one of a series of siRNAs targeting pattern recognition receptors. Violin plots indicate median, lower and upper quartiles; N = 6 values from individual wells, representative of n = 3 biologically-independent experiments, two-way ANOVA post hoc Dunnett’s test. P values, ****< 0.0001. I Western blot of SUM149 BRCA1 -Mut and BRCA1 -Rev cells subjected to co-transfection with ADAR1 siRNA and one of a series of siRNAs targeting pattern recognition receptors. Data representative of n = 3 biologically-independent experiments. J , K Cell survival of SUM149 BRCA1 -Mut and BRCA1 -Rev cells transfected with a concentration range (nM) of ADAR1 siRNA in the context of exposure to the JAK/STAT pathway inhibitors (JSPi) ruxolitinib ( J ; 10 µM) or upadacitinib ( K ; 32 µM). Box-and-whiskers indicate median, lower and upper quartiles, and the min to max range; N = 4 values from individual wells, representative of n = 3 biologically-independent experiments, two-way ANOVA post hoc Dunnett’s test. P values, ****< 0.0001. siCTRL, non-targeting, negative control siRNA; siPLK1, PLK1 -targeting, positive control siRNA. Source data are provided as a Source Data file.

Article Snippet: The JAK/STAT pathway inhibitors ruxolitinib (INCB-18424; JAK1/2i), upadacitinib (ABT-494; JAK1i) and deucravacitinib (BMS-986165, TYK2i), and the integrated stress response inhibitor ISRIB were purchased from MedChemExpress.

Techniques: Western Blot, Transduction, shRNA, Titration, Quantitative RT-PCR, Transfection, Concentration Assay, Gene Expression, Mutagenesis, Cotransfection, Negative Control, Positive Control