Abstract
Background
Targeting DNA damage repair factors, such as DNA-dependent protein kinase catalytic subunit (DNA-PKcs), may offer an opportunity for effective treatment of multiple myeloma (MM). In combination with DNA damage-inducing agents, this strategy has been shown to improve chemotherapies partially via activation of cGAS-STING pathway by an elevated level of cytosolic DNA. However, as cGAS is primarily sequestered by chromatin in the nucleus, it remains unclear how cGAS is released from chromatin and translocated into the cytoplasm upon DNA damage, leading to cGAS-STING activation.
Methods
We examined the role of DNA-PKcs inhibition on cGAS-STING-mediated MM chemosensitivity by performing mass spectrometry and mechanism study.
Results
Here, we found DNA-PKcs inhibition potentiated DNA damage-inducing agent doxorubicin-induced anti-MM effect by activating cGAS-STING signaling. The cGAS-STING activation in MM cells caused cell death partly via IRF3-NOXA-BAK axis and induced M1 polarization of macrophages. Moreover, this activation was not caused by defective classical non-homologous end joining (c-NHEJ). Instead, upon DNA damage induced by doxorubicin, inhibition of DNA-PKcs promoted cGAS release from cytoplasmic chromatin fragments and increased the amount of cytosolic cGAS and DNA, activating cGAS-STING.
Conclusions
Inhibition of DNA-PKcs could improve the efficacy of doxorubicin in treatment of MM by de-sequestrating cGAS in damaged chromatin.
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Data availability
The raw RNA-seq datasets for this study are available from the NCBI Sequence Read Archive (SRA) with the accession number code PRJNA984948. Flow cytometry datasets are publicly accessible at the repository Zenodo (https://doi.org/10.5281/zenodo.8042604). Source data for the figures and supplementary figures are provided as a Source Data file. Source Data are provided with this paper.
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Acknowledgements
We thank members of the Cai lab and the **e lab for helpful discussions. We also thank Bi Chao and Hong **aoli from the Core Facilities, Zhejiang University School of Medicine for their technical support.
Funding
This work was supported by the National Natural Science Foundation of China (82100212 to MMD, U22A20291 to ZC, No. 31870806 to AYX and 32071439 to YLF), the Department of Science and Technology of Hangzhou (202204A05 to AYX), the Zhejiang Provincial Natural Science Foundation of China (LQ21H160015 to EFZ and LZ22H160009 to JSH) and the Key Research and Development Plan Project of Zhejiang Province (2020C03014 to ZC).
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ZC, AYX, MMD and JNZ initiated and designed the study. JNZ, MMD and WC performed experiments. JNZ, MMD, YLF and AYX analyzed experimental results. AYX and JNZ wrote the paper. HGC performed the bioinformatics analyses. HYG, EFZ, YLF and SCL provided assistance and valuable advice on the manuscript. JSH, AYX and ZC supervised the experiments.
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All experimental methods were carried out in accordance with relevant guidelines such as the Declaration of Helsinki. All procedures involving animals were in accordance with the Animal Experimental Ethical Committee of the First Affiliated Hospital, School of Medicine, Zhejiang University (Reference Number: 2022-1578). Human samples-based study was approved by the Research Ethics Committee of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China (Reference Number: 2022-930).
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Zhang, JN., Dong, MM., Cao, W. et al. Disruption of DNA-PKcs-mediated cGAS retention on damaged chromatin potentiates DNA damage-inducing agent-induced anti-multiple myeloma activity. Br J Cancer (2024). https://doi.org/10.1038/s41416-024-02742-3
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DOI: https://doi.org/10.1038/s41416-024-02742-3
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