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

figure

posted on 2024-02-26, 14:20 authored by Lisa Choy, Stephen Norris, Xiumin Wu, Ganesh Kolumam, Ari Firestone, Jeffrey Settleman, David Stokoe

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.

Funding

Calico LLC

History

ARTICLE ABSTRACT

Type I IFN signaling is a crucial component of antiviral immunity that has been linked to promoting the efficacy of some chemotherapeutic drugs. We developed a reporter system in HCT116 cells that detects activation of the endogenous IFI27 locus, an IFN target gene. We screened a library of annotated compounds in these cells and discovered Aurora kinase inhibitors (AURKi) as strong hits. Type I IFN signaling was found to be the most enriched gene signature after AURKi treatment in HCT116, and this signature was also strongly enriched in other colorectal cancer cell lines. The ability of AURKi to activate IFN in HCT116 was dependent on MAVS and RIG-I, but independent of STING, whose signaling is deficient in these cells. MAVS dependence was recapitulated in other colorectal cancer lines with STING pathway deficiency, whereas in cells with intact STING signaling, the STING pathway was required for IFN induction by AURKi. AURKis were found to induce expression of endogenous retroviruses (ERV). These ERVs were distinct from those induced by the DNA methyltransferase inhibitors (DNMTi), which can induce IFN signaling via ERV induction, suggesting a novel mechanism of action. The antitumor effect of alisertib in mice was accompanied by an induction of IFN expression in HCT116 or CT26 tumors. CT26 tumor growth inhibition by alisertib was absent in NSG mice versus wildtype (WT) mice, and tumors from WT mice with alisertib treatment showed increased in CD8+ T-cell infiltration, suggesting that antitumor efficacy of AURKi depends, at least in part, on an intact immune response. Some cancers deactivate STING signaling to avoid consequences of DNA damage from aberrant cell division. The surprising activation of MAVS/RIG-I signaling by AURKi might represent a vulnerability in STING signaling deficient cancers.