CDK9 degrader induces BRCAness and sensitizes castration-resistant prostate cancer to PARP inhibitor.

Abstract

Rationale: Castration-resistant prostate cancer (CRPC) poses significant therapeutic challenges due to its aggressive nature and limited effective treatments. Although PARP inhibitor olaparib has been approved for metastatic CRPC patients bearing BRCA1/2 mutations, its application is confined to this specific patient subpopulation. The induction of "BRCAness" feature in CRPC patients without BRCA1/2 mutations becomes a significant challenge.

Methods: A transcriptomic analysis to identify potential "BRCAness" regulator was performed on 921 prostate cancer (PCa) patients from 6 public datasets and validated in our own cohort. D45, a selective small-molecule protein degrader for cyclin-dependent kinase 9 (CDK9), alone or combined with olaparib, was applied in CRPC cell lines (C4-2 and 22Rv1) lacking BRCA1/2 mutations for cell viability, colony formation and apoptosis-related assays. "BRCAness" phenotype was characterized by Western blotting and γH2AX foci accumulation assays. RNA-seq and CUT&Tag assays were used to reveal how D45 regulated homologous recombination repair (HRR)-related genes.

Results: We found the transcriptional regulator CDK9 was overexpressed in CRPC and correlated with advanced Gleason scores, metastasis, and poor prognosis. D45 treatment decreased cell survival, and led to downregulation of HRR-related genes (BRCA1/2 and RAD51) with the reduced recruitment of phosphorylated-RNA polymerase II (pSer2) to the ends of these genes with increased DNA damage, indicating a "BRCAness" phenotype induction. The synthetic vulnerability synergized with D45 plus olaparib was thus tested in vitro, showing the enhanced apoptosis in CRPC cells. Moreover, sequential D45 and olaparib administration significantly suppressed 22Rv1 xenograft growth in vivo (P < 0.001), reduced RAD51 expression and increased DNA damage. Toxicity was tolerable and consistent with prior reports. Their synergistic effect was confirmed in ex vivo explants from human PCa specimens.

Conclusions: Our findings demonstrated CDK9 as a master regulator of BRCAness and proposed targeting CDK9 as a potential strategy to sensitize CRPC patients without BRCA1/2 mutations to PARP inhibition.