DNA repair gene mutations are frequent in castration-resistant prostate cancer (CRPC), suggesting eligibility for poly(ADP-ribose) polymerase inhibitor (PARPi) treatment. However, therapy resistance is a major clinical challenge and genes contributing to PARPi resistance are poorly understood. Using a genome-wide CRISPR-Cas9 knockout screen, this study aimed at identifying genes involved in PARPi resistance in CRPC. Based on the screen, we identified PARP1, and six novel candidates associated with olaparib resistance upon knockout. For validation, we generated multiple knockout populations/clones per gene in C4 and/or LNCaP CRPC cells, which confirmed that loss of PARP1, ARH3, YWHAE, or UBR5 caused olaparib resistance. PARP1 or ARH3 knockout caused cross-resistance to other PARPis (veliparib and niraparib). Furthermore, PARP1 or ARH3 knockout led to reduced autophagy, while pharmacological induction of autophagy partially reverted their PARPi resistant phenotype. Tumor RNA sequencing of 126 prostate cancer patients identified low ARH3 expression as an independent predictor of recurrence. Our results advance the understanding of PARPi response by identifying four novel genes that contribute to PARPi sensitivity in CRPC and suggest a new model of PARPi resistance through decreased autophagy.
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This work was supported by grants from The Novo Nordisk Foundation (KDS), Aase & Ejnar Danielsens Fond (MBI), Fabrikant Einar Willumsens Mindelegat (MBI), Tømrermester Jørgen Holm & Hustru Elisa F. Hansens Mindelegat (MBI), Direktør Emil C. Hertz og Hustru Inge Hertz Fond (MBI), Graduate School of Health (Aarhus University; MBI), Helge Peetz & Verner Peetz & Hustru Vilma Peetz Fond (MBI), Beckett Fonden (MBI), and NEYE Fonden (MBI). The Danish Cancer Biobank is acknowledged for providing patient samples and information on handling and storage.
The authors declare no competing interests.
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Ipsen, M.B., Sørensen, E.M.G., Thomsen, E.A. et al. A genome-wide CRISPR-Cas9 knockout screen identifies novel PARP inhibitor resistance genes in prostate cancer. Oncogene 41, 4271–4281 (2022). https://doi.org/10.1038/s41388-022-02427-2