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Blocking LBH expression causes replication stress and sensitizes triple-negative breast cancer cells to ATR inhibitor treatment

Oncogene volume 43, pages 851–865 (2024)Cite this article

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Abstract

Triple-negative (ER-PR-HER2-) breast cancers (TNBC) are highly aggressive and difficult to treat. TNBC exhibit high genomic instability, which enables them to adapt and become resistant to chemo/radiation therapy, leading to rapid disease relapse and mortality. The pro-survival factors that safeguard genome integrity in TNBC cells are poorly understood. LBH is an essential mammary stem cell-specific transcription regulator in the WNT pathway that is aberrantly overexpressed in TNBC, correlating with poor prognosis. Herein, we demonstrate a novel role for LBH in promoting TNBC cell survival. Depletion of LBH in multiple TNBC cell models triggered apoptotic cell death both in vitro and in vivo and led to S-G2M cell cycle delays. Mechanistically, LBH loss causes replication stress due to DNA replication fork stalling, leading to ssDNA breaks, ɣH2AX and 53BP1 nuclear foci formation, and activation of the ATR/CHK1 DNA damage response. Notably, ATR inhibition in combination with LBH downmodulation had a synergistic effect, boosting TNBC cell killing and blocking in vivo tumor growth. Our findings demonstrate, for the first time, that LBH protects the genome integrity of cancer cells by preventing replicative stress. Importantly, they uncover new synthetic lethal vulnerabilities in TNBC that could be exploited for future multi-modal precision medicine.

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Fig. 1: LBH silencing in TNBC cells induces apoptotic cell death.
Fig. 2: LBH silencing causes S-G2/M cell cycle delays and DNA replication stalling.
Fig. 3: LBH silencing in TNBC cells increases genome instability and DNA damage.
Fig. 4: LBH silencing activates the ATR/CHK1 DNA damage response.
Fig. 5: ATR inhibition synergizes with LBH knockdown to promote TNBC cell killing.
Fig. 6: ATR inhibition exacerbates DNA damage induced by LBH loss.
Fig. 7: LBH depletion synergizes with ATR inhibition to suppress in vivo TNBC tumor growth.
Fig. 8: LBH downmodulation enhances the efficacy of ATRi in other cancer types.

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Data availability

All relevant data are available from the authors upon request.

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Acknowledgements

We thank the Analytic Imaging Core of the Miami Project to Cure Paralysis for confocal imaging. This work was supported by NIH/NIGMS Grant R01GM113256 (K.J.B.), the Department of Defense (DoD)/Breast Cancer Research Program (BCRP) Breakthrough Award W81XWH-19-1-0255 (K.J.B.); a Tumor Biology Program Trainee Award (K.G.), and research funds from the Sylvester Comprehensive Cancer Center (K.J.B.). Research reported in this publication was performed in part at the Flow Cytometry Shared Resource (FCSR; RRID: SCR_022501) and the Cancer Modeling Shared Resource (CMSR; RRID: SCR_022889) of the Sylvester Comprehensive Cancer Center at the University of Miami Miller School of Medicine, which is supported by the National Cancer Institute Cancer Center Support Grant (CCSG) P30-CA240139.

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Authors and Affiliations

  1. DeWitt Daugherty Department of Surgery, Molecular Oncology Program, University of Miami Miller School of Medicine, Miami, FL, USA

    Koteswararao Garikapati, In-Chi Young, Sunhwa Hong & Karoline J. Briegel

  2. Braman Family Breast Cancer Institute at the Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, FL, USA

    Koteswararao Garikapati, In-Chi Young, Sunhwa Hong & Karoline J. Briegel

  3. Department of Radiation Oncology and Tumor Biology Program at the Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, FL, USA

    Priyamvada Rai

  4. Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, FL, USA

    Chaitanya Jain

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  1. Koteswararao Garikapati
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Contributions

K.G. and K.J.B. conceived the study and designed the experiments. K.G. performed most experiments, analyzed the data, prepared the figures, and wrote the first draft of the manuscript. I.-C.Y. and S.H. generated stable LBH knockdown cell lines. I.-C.Y. performed in vivo Xenograft experiments and data analysis. P.R. provided essential reagents and feedback on the study. C.J. reviewed and edited the manuscript. K.J.B. directed the study, reviewed the data, wrote the manuscript, and secured funding.

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Correspondence to Karoline J. Briegel.

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Garikapati, K., Young, IC., Hong, S. et al. Blocking LBH expression causes replication stress and sensitizes triple-negative breast cancer cells to ATR inhibitor treatment. Oncogene 43, 851–865 (2024). https://doi.org/10.1038/s41388-024-02951-3

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