Tumor-derived neutrophil extracellular trap–associated DNA impairs treatment efficacy in breast cancer via CCDC25-dependent epithelial-mesenchymal transition
Heliang Li, Yetong Zhang, Jianghua Lin, Jiayi Zeng, Xinyan Liang, Linxi Xu, Jiang Li, Xiaoming Zhong, Xu Liu, Zhou Liu, Xinyu Yang, Yunyi Zhang, Shun Wang, Erwei Song, Man Nie, Linbin Yang
Heliang Li, Yetong Zhang, Jianghua Lin, Jiayi Zeng, Xinyan Liang, Linxi Xu, Jiang Li, Xiaoming Zhong, Xu Liu, Zhou Liu, Xinyu Yang, Yunyi Zhang, Shun Wang, Erwei Song, Man Nie, Linbin Yang
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Research Article Immunology Oncology

Tumor-derived neutrophil extracellular trap–associated DNA impairs treatment efficacy in breast cancer via CCDC25-dependent epithelial-mesenchymal transition

Abstract

Neutrophil extracellular traps (NETs) are associated with cancer progression; however, the functional role and clinical importance of NET-DNA in therapeutic resistance remain unclear. Here, we show that chemotherapy and radiotherapy provoke NET-DNA formation in primary tumor and metastatic organs in breast cancer patients and mouse models, and the level of NET-DNA correlates with treatment resistance. Mechanistically, the cathepsin C in tumor debris generated by anticancer therapy is phagocytosed by macrophages and drives CXCL1/2 and complement factor B production via activating the TLR4/NF-κB signaling pathway, subsequently promoting NETosis and impairing therapeutic efficacy. Importantly, we demonstrate that NET-DNA sensor CCDC25 is indispensable in NET-mediated treatment resistance by inducing cancer cell epithelial-mesenchymal transition via pyruvate kinase isoform M2–mediated STAT3 phosphorylation. Clinically, tumoral CCDC25 abundance is closely associated with poor prognosis in patients who underwent chemotherapy. Overall, our data reveal the mechanism of NET formation and elucidate the interaction of NET-CCDC25 in therapy resistance, highlighting CCDC25 as an appealing target for anticancer interventions.

Authors

Heliang Li, Yetong Zhang, Jianghua Lin, Jiayi Zeng, Xinyan Liang, Linxi Xu, Jiang Li, Xiaoming Zhong, Xu Liu, Zhou Liu, Xinyu Yang, Yunyi Zhang, Shun Wang, Erwei Song, Man Nie, Linbin Yang

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Figure 4

Macrophages promote CXCL1/2 and CFB elevation and NETosis via phagocytosis of treatment-induced tumor debris.

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Macrophages promote CXCL1/2 and CFB elevation and NETosis via phagocytos...
(A) GSEA showing functional enrichment of KEGG pathways in macrophage subset from scRNA-Seq of 4T1 tumors (permutation test). (B) GSEA showing functional enrichment of KEGG pathways in RNA-Seq of patient breast tumors post-chemotherapy versus pretreatment (GSE18728; pre: n = 21, post: n = 22; permutation test). (C and D) Representative immunofluorescence images (C) and quantification of the proportions of Luci+F4/80+ cells (D) in tumors, livers, and lungs (n = 5/group). Luci, luciferase. * indicates areas magnified in the top-left insets. Tumor and liver, unpaired t test. Lung, Mann-Whitney test. (E) Quantification of the proportions of GFP+F4/80+ cells in MCF-7 tumors (n = 5/ group, unpaired t test). (F) Quantification of the proportions of tumoral PyMT+F4/80+ cells in MMTV-PyMT FVB/NJ mice (n = 5/group, Mann-Whitney test). (G) ELISA levels of CXCL1/2 and CFB in culture medium of BMDMs with indicated treatments (n = 3/group, 1-way ANOVA with Tukey’s test). (H) Representative immunofluorescence images in 4T1 tumors after chemotherapy. White boxes indicate areas magnified in the next row. Data represent 3 independent experiments. (I and J) 4T1 cells were injected alone or coinjected with 4T1 debris orthotopically in BALB/c mice (n = 5/group). (I) Representative images and quantification of tumoral NETs. (J) Quantification of tumoral CXCL1+F4/80+, CXCL2+F4/80+, and CFB+F4/80+ cells (Mann-Whitney test). Scale bar, 20 μm. (K–M) Orthotopic 4T1 tumors treated with Dox together with UNC2250 or vehicle (n = 5/group). (K) Tumor growth curves. (L) Quantification of NETs. (M) Quantification of CXCL1+F4/80+, CXCL2+F4/80+, and CFB+F4/80+ cell counts. Data were analyzed by 2-way repeated measures ANOVA with Šídák’s test and compared on day 29 (K), or unpaired t test (L and M). Data represent mean ± SD. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05.