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DNA damage induces STING mediated IL-6-STAT3 survival pathway in triple-negative breast cancer cells and decreased survival of breast cancer patients

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Abstract

Triple-negative breast cancer is aggressive and metastatic breast cancer type and shows immune evasion, drug resistance, relapse and poor survival. Anti-cancer therapy like ionizing radiation and chemotherapeutic drug majorly induces DNA damage hence, alteration in DNA damage repair and downstream pathways may contribute to tumor cell survival. DNA damage during chemotherapy is sensed by cyclic GMP-AMP synthase(cGAS)-stimulator of interferon genes (STING), which determines the anti-tumor immune response by modulating the expression of programmed cell death ligand-1 (PD-L1), immune suppressor, in the tumor microenvironment. Triple-negative breast cancer cells are cGAS-STING positive and modulation of this pathway during DNA damage response for survival and immune escape mechanism is not well understood. Here we demonstrate that doxorubicin-mediated DNA damage induces STING mediated NF-κB activation in triple-negative as compared to ER/PR positive breast cancer cells. STING-mediated NF-κB induces the expression of IL-6 in triple-negative breast cancer cells and activates pSTAT3, which enhances cell survival and PD-L1 expression. Doxorubicin and STAT3 inhibitor act synergistically and inhibit cell survival and clonogenicity in triple-negative breast cancer cells. Knockdown of STING in triple-negative breast cancer cells enhances CD8 mediated immune cell death of breast cancer cells. The combinatorial treatment of triple-negative breast cells with doxorubicin and STAT3 inhibitor reduces PD-L1 expression and activates immune cell-mediated cancer cell death. Further STING and IL-6 levels show a positive correlation in breast cancer patients and poor survival outcomes. The study here strongly suggests that STING mediated activation of NF-κB enhances IL-6 mediated STAT3 in triple-negative breast cancer cells which induces cell survival and immune-suppressive mechanism.

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The data that support the findings of this study are available on request from the corresponding author.

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Acknowledgements

This work was partially supported by the Department of Science and Technology, Govt. of India, Grant Number INT/Korea/P-39 to Prof. Rajesh Singh. Authors also acknowledged pratical support from GSBTM, GJ, India The authors acknowledge the instrumentation facility at the Department of Biochemistry, The M. S. University of Baroda, Vadodara under the DST FIST program. DBT BUILIDER program supported by Department of Biotechnology, Govt of India. Minal Mane received her fellowship from CSIR, Govt. of India. Dhruv Gohel received his fellowship from ICMR, India. Fatema Currim received her fellowship from the DST-INSPIRE program, India. Anjali Shinde is ICMR SRF.

Funding

Department of Science and Technology, Govt. of India, Grant Number INT/Korea/P-39 to Prof. Rajesh Singh.

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat, 390002, India

    Hitesh Vasiyani, Minal Mane, Anjali Shinde, Jyoti Singh, Dhruv Gohel, Fatema Currim & Rajesh Singh

  2. Department of Microbiology & Biotechnology Center, The M.S. University of Baroda, Vadodara, Gujarat, 390002, India

    Khushboo Rana & Ratika Srivastava

  3. Institute for Cell Engineering, John Hopkins University School of Medicine, 733 North Broadway, MRB 731, Baltimore, MD, 21205, USA

    Milton Roy

  4. Department of Mol and Human Genetics, Banaras Hindu University, Varanasi, UP, 221005, India

    Rajesh Singh

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  1. Hitesh Vasiyani
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Contributions

HV: conceptualization, methodology, formal analysis, investigation, writing—original draft, AS: data curation, writing—review and editing, resources; MR: writing—review and editing, resources; MM: resources; JS: resources; DG: resources; FC: resources; KR: data curation, resources; RS: resources; RS: funding acquisition, project administration, supervision, conceptualization, writing—review and editing.

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Correspondence to Rajesh Singh.

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Vasiyani, H., Mane, M., Rana, K. et al. DNA damage induces STING mediated IL-6-STAT3 survival pathway in triple-negative breast cancer cells and decreased survival of breast cancer patients. Apoptosis 27, 961–978 (2022). https://doi.org/10.1007/s10495-022-01763-8

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