FIGURE 3 from Inhibition of Aurora Kinase Induces Endogenous Retroelements to Induce a Type I/III IFN Response via RIG-I

Aurora kinase inhibitors activate IFN signaling. A, Heat map of RNA-seq data from the HCT116-IFI27 reporter line treated for 5 days with 100 nmol/L decitabine, 1 µmol/L 5-azacytidine, 1 µmol/L alisertib, 1 µmol/L tozasertib, 830 nmol/L foretinib, 2.5 µmol/L axitinib, 2.5 µmol/L irinotecan, 1 µmol/L selumetib, 830 nmol/L binimetinib, or an equivalent volume of DMSO. Heat map shows all significant genes using a Padj < 1e-10 in at least one condition. B, Top 20 most induced genes after alisertib treatment in the RNA-seq data shown in A, filtered for qval ≤ 0.05. Yellow colored genes are members of the Hallmark IFN Alpha gene set. C, GSEA hallmark IFN alpha gene set enrichment plot of RNA-seq data shown in A resulting from alisertib treatment. D, Summary of all positively or negatively enriched hallmark GSEA from the RNA-seq data after either alisertib or decitabine treatments, with a cutoff of FDR qval of <0.05. NES = normalized enrichment score. IFNα is the most highly induced of the enriched gene sets after AURK inhibition. E, Colorectal cancer cell lines were treated for 5 days with AURKi (1 µmol/L alisertib except for CT26, 1 µmol/L tozasertib) or the DNMTi decitabine (200 nmol/L), then RNA-seq performed and subjected to GSEA. GSEA normalized enrichment scores (NES) for the hallmark interferon alpha gene set are indicated along with the corresponding q values for each analysis. n/a = not applicable. F, qPCR measurement of expression of IFNβ, IFNλ1, and IFNλ2 across the human colorectal cancer cell line panel after 5 days treatment with 1 µmol/L alisertib or 100 nmol/L barasertib. Values are relative to HT-29 with DMSO control treatment. Significance shown for each treatment versus DMSO for each cell line; n.d = not detected. G, CT26 cells were treated with DMSO, 10 µmol/L ruxolitinib, 1 µmol/L alisertib, or 200 nmol/L decitabine, and RNA collected at 24 hours, 48 hours, 72 hours, or 5 days. qPCR was performed to illustrate induction of Ifnβ and target genes Ifit1, Ifit2, and Cxcl10. Significance is shown for comparison between DMSO versus decitabine or alisertib for each timepoint.