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Evaluating the therapeutic potential of ADAR1 inhibition for triple-negative breast cancer

A Correction to this article was published on 09 February 2021

This article has been updated

Abstract

Triple-negative breast cancer (TNBC) is the deadliest form of breast cancer. Unlike other types of breast cancer that can be effectively treated by targeted therapies, no such targeted therapy exists for all TNBC patients. The ADAR1 enzyme carries out A-to-I editing of RNA to prevent sensing of endogenous double-stranded RNAs. ADAR1 is highly expressed in breast cancer including TNBC. Here, we demonstrate that expression of ADAR1, specifically its p150 isoform, is required for the survival of TNBC cell lines. In TNBC cells, knockdown of ADAR1 attenuates proliferation and tumorigenesis. Moreover, ADAR1 knockdown leads to robust translational repression. ADAR1-dependent TNBC cell lines also exhibit elevated IFN stimulated gene expression. IFNAR1 reduction significantly rescued the proliferative defects of ADAR1 loss. These findings establish ADAR1 as a novel therapeutic target for TNBC tumors.

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Fig. 1: ADAR1 is highly expressed in all breast cancer subtypes.
Fig. 2: ADAR1 is required for TNBC survival and proliferation.
Fig. 3: ADAR1 is required for TNBC transformation and tumorigenesis.
Fig. 4: PKR is overexpressed in TNBC and activated upon ADAR loss.
Fig. 5: ADAR1-dependent TNBCs exhibit elevated ISG expression and INFAR1 loss rescues ADAR1-knockdown phenotype.

Data and code availability

CCLE RNA-seq count data (CCLE_RNAseq_genes_counts_20180929.gct.gz, CCLE_RNAseq_rsem_transcripts_tpm_20180929.txt.gz) were obtained from the Broad Institute Cancer Cell Line Encyclopedia and is available at https://portals.broadinstitute.org/ccle/data. Dependency data (D2_combined_gene_dep_scores.csv, Achilles_gene_effect.csv) were obtained from Broad Institute DepMap Portal and is available at https://depmap.org/portal/download/. TCGA breast cancer RNA-seq (illuminahiseq_rnaseqv2-RSEM_genes, illuminahiseq_rnaseqv2-RSEM_isoforms_normalized) and clinical data (Merge_Clinical) were obtained from the Broad Institute FireBrowse and are available at http://firebrowse.org/. All custom R scripts used in this study are available on GitHub (https://github.com/cottrellka/ADAR_TNBC). Lentiviral production and transduction; flow cytometric analysis of apoptosis; cell proliferation and focus formation assays; soft agar transformation assay; polysome profiling; immunohistochemistry. These experiments were performed as previously described, and further details can be found in the Supplementary Information [46, 51, 52].

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Acknowledgements

This work was supported by R01CA190986 (JDW), F32GM131514 (KAC), and TL1TR002344 (C-PK) from the National Institute of Health, and W81XWH-18-1-0025 from the Department of Defense (JDW). This work was supported by the Longer Life Foundation: A RGA/Washington University partnership. We thank Kazuko Nishikura (The Wistar Institute) for providing ADAR1 expressing constructs. The results shown here are in whole or part based upon data generated by the TCGA Research Network: https://www.cancer.gov/tcga.

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Conceptualization: C-PK, KAC, and JDW; methodology: C-PK, KAC, and JDW; software: KAC; investigation: C-PK, KAC, SR, ERB, RDK, EAB, ECF, TS, LM, and JDW; writing—original draft: C-PK and KAC; writing—review and editing: C-PK, KAC, SR, ERB, RDK, LM, and JDW; funding acquisition: JDW; supervision: JDW.

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Correspondence to Jason D. Weber.

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The original online version of this article was revised: The word “orthopedic” was changed to “orthotopic” in subheading “Mammary gland orthotopic implantation” and in the first sentence of section “Mammary gland orthotopic implantation”.

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Kung, CP., Cottrell, K.A., Ryu, S. et al. Evaluating the therapeutic potential of ADAR1 inhibition for triple-negative breast cancer. Oncogene 40, 189–202 (2021). https://doi.org/10.1038/s41388-020-01515-5

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