Jab1 regulates HRR mRNA stability to modulate PARP inhibitor sensitivity in triple-negative breast cancer

Triple-negative breast cancer (TNBC) is a highly aggressive breast cancer subtype associated with the highest mortality rate among all breast cancer subtypes, primarily due to the absence of actionable therapeutic targets. Although poly (ADP-ribose) polymerase inhibitors (PARPi) have shown promising therapeutic effects in TNBC patients harboring homologous recombination deficiency (HRD), their clinical benefit remains limited, highlighting an urgent need for novel targets that enhance PARPi efficacy. This study investigates the role of Jab1 in regulating the stability of homologous recombination repair (HRR)-related RNAs and evaluates its potential as a therapeutic target to enhance PARPi sensitivity in TNBC.

RNA-Seq analysis revealed that shRNA-mediated Jab1 knockdown profoundly affected HRR and DNA replication processes in TNBC cells. Using Nuclear Run-On Assay, RNA Immunoprecipitation, RNA Pull-Down Assay, and RIP-Seq, we identified Jab1 as a potential RNA-binding protein (RBP) that stabilizes HRR-related mRNAs by competing with the exosome complex. Genetic and pharmacological inhibition of Jab1 (using CSN5i-3) were evaluated for their impact on HRR efficiency, ionizing radiation (IR) sensitivity, and PARPi sensitivity. A comprehensive panel of in vitro assays was performed, including clonogenic survival assays, PrestoBlue assays, apoptosis assays, DR-GFP reporter assays, qRT-PCR, Western blot, comet assays, and immunofluorescence. In vivo efficacy was assessed using zebrafish xenografts, nude mouse xenografts, and syngeneic orthotopic mouse models to examine the therapeutic effect of Jab1 inhibition in combination with PARPi.

Jab1 was found to be overexpressed in TNBC and correlated with poor clinical outcomes. Functional analyses revealed that Jab1 knockdown impaired HRR, increased DNA damage accumulation, and sensitized TNBC cells to IR and PARPi, irrespective of BRCA mutation status. Mechanistically, Jab1 functioned as an RBP through its MPN domain, stabilizing HRR-related transcripts by competitively antagonizing the RNA exosome complex. Pharmacological inhibition of Jab1 using CSN5i-3 recapitulated these effects and synergized with PARPi to induce synthetic lethality. In multiple preclinical models, this combination significantly suppressed tumor growth and promoted apoptosis.

This study uncovers a novel role for Jab1 as an RBP, specifically through interactions between its MPN domain and HRR-related RNAs, regulating RNA stability and maintaining HRR competency. Targeting Jab1 represents a promising strategy to pharmacologically induce HRD and enhance the efficacy of PARPi therapies in TNBC. This combination approach may hold translational value for improving clinical outcomes in patients with TNBC.