5-HT orchestrates histone serotonylation and citrullination to drive neutrophil extracellular traps and liver metastasis
Kaiyuan Liu, Yingchao Zhang, Genyu Du, Xinyu Chen, Lingling Xiao, Luyao Jiang, Na Jing, Penghui Xu, Chaoxian Zhao, Yiyun Liu, Huifang Zhao, Yujiao Sun, Jinming Wang, Chaping Cheng, Deng Wang, Jiahua Pan, Wei Xue, Pengcheng Zhang, Zhi-Gang Zhang, Wei-Qiang Gao, Shu-Heng Jiang, Kai Zhang, Helen He Zhu
Kaiyuan Liu, Yingchao Zhang, Genyu Du, Xinyu Chen, Lingling Xiao, Luyao Jiang, Na Jing, Penghui Xu, Chaoxian Zhao, Yiyun Liu, Huifang Zhao, Yujiao Sun, Jinming Wang, Chaping Cheng, Deng Wang, Jiahua Pan, Wei Xue, Pengcheng Zhang, Zhi-Gang Zhang, Wei-Qiang Gao, Shu-Heng Jiang, Kai Zhang, Helen He Zhu
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Research Article Cell biology Oncology

5-HT orchestrates histone serotonylation and citrullination to drive neutrophil extracellular traps and liver metastasis

Abstract

Serotonin (5-HT) is a neurotransmitter that has been linked to tumorigenesis. Whether and how 5-HT modulates cells in the microenvironment to regulate tumor metastasis is largely unknown. Here, we demonstrate that 5-HT was secreted by neuroendocrine prostate cancer (NEPC) cells to communicate with neutrophils and to induce the formation of neutrophil extracellular traps (NETs) in the liver, which in turn facilitated the recruitment of disseminated cancer cells and promoted liver metastasis. 5-HT induced histone serotonylation (H3Q5ser) and orchestrated histone citrullination (H3cit) in neutrophils to trigger chromatin decondensation and facilitate the formation of NETs. Interestingly, we uncovered in this process a reciprocally reinforcing effect between H3Q5ser and H3cit and a crosstalk between the respective writers enzyme transglutaminase 2 (TGM2) and peptidylarginine deiminase 4 (PAD4). Genetic ablation or pharmacological targeting of TGM2, or inhibition of the 5-HT transporter (SERT) with the FDA-approved antidepressant drug fluoxetine reduced H3Q5ser and H3cit modifications, suppressed NET formation, and effectively inhibited NEPC, small-cell lung cancer, and thyroid medullary cancer liver metastasis. Collectively, the 5-HT–triggered production of NETs highlights a targetable neurotransmitter/immune axis that drives liver metastasis of NE cancers.

Authors

Kaiyuan Liu, Yingchao Zhang, Genyu Du, Xinyu Chen, Lingling Xiao, Luyao Jiang, Na Jing, Penghui Xu, Chaoxian Zhao, Yiyun Liu, Huifang Zhao, Yujiao Sun, Jinming Wang, Chaping Cheng, Deng Wang, Jiahua Pan, Wei Xue, Pengcheng Zhang, Zhi-Gang Zhang, Wei-Qiang Gao, Shu-Heng Jiang, Kai Zhang, Helen He Zhu

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Figure 2

NEPC-derived 5-HT potentiates the formation of NETs and liver metastasis.

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NEPC-derived 5-HT potentiates the formation of NETs and liver metastasis...
(A and B) IF staining images (A) and quantification results (B) showing NET formation in murine BMDNs in response to 5-HT or vehicle control treatment (n = 10 biological replicates). PMA-induced NET formation and served as a positive control. Scale bars: 50 μm. (C and D) IF staining images (C) and quantification results (D) showing NET formation in murine BMDNs upon the treatment of CM collected from scrambled- and shTph1-infected Rb1Δ/Δ Trp53Δ/Δ organoids (n = 9 biological replicates). Scale bars: 50 μm. (E and F) IF images (E) and quantification (F) of the NET-forming capacity of human PBDNs upon 5-HT or vehicle control treatment (n = 10 biological replicates). Scale bars: 50 μm. (G) MNase digestion assay showing that addition of 5-HT induced decondensed NET chromatin in HL-60 granulocytes compared with calcium ionophore treatment. (H–J) In vivo experiments demonstrating a reduction of liver metastatic burden (H) in shTph1-infected versus scrambled shRNA–treated mice, and a regained liver metastatic burden in 5-HTP–treated, shTph1–infected Rb1Δ/Δ Trp53Δ/Δ organoid–implanted mice via i.v. injection (n = 7 mice, each group). H&E staining (I) and quantification data (J) validated the significant decrease in liver metastatic foci in Tph1-KD Rb1Δ/Δ Trp53Δ/Δ organoid–inoculated mice and regained liver metastatic foci numbers in 5-HTP–treated mice. Scale bars: 2 mm. (K and L) IF images of liver sections (K) and quantification results (L) revealing a significant decline in NET formation in Tph1-KD Rb1Δ/Δ Trp53Δ/Δ organoid–inoculated mice (n = 7 mice, each group). *P < 0.05, **P < 0.01, and ***P < 0.001, by Kruskal-Wallis test followed by Dunnett’s test (B, D, F, and L) and 1-way ANOVA followed by Tukey’s test (J). Data are presented as the mean ± SEM.