J Cancer 2022; 13(4):1261-1271. doi:10.7150/jca.64170 This issue

Research Paper

Neutrophil extracellular traps in hepatocellular carcinoma are enriched in oxidized mitochondrial DNA which is highly pro-inflammatory and pro-metastatic

Lu-Yu Yang1,2,3#, Xiao-Tian Shen1,2,3#, Hao-Ting Sun1,2,3, Wen-Wei Zhu1,2,3, Ju-Bo Zhang3,4, Lu Lu1,2,3✉

1. Department of General Surgery, Huashan Hospital, Fudan University, Shanghai, China
2. Caner Metastasis Institute, Fudan University, Shanghai, China
3. Liver disease center, Huashan Hospital, Fudan University, Shanghai, China
4. Department of Infection Disease, Huashan Hospital, Fudan University, Shanghai, China
#Co-first authors

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Citation:
Yang LY, Shen XT, Sun HT, Zhu WW, Zhang JB, Lu L. Neutrophil extracellular traps in hepatocellular carcinoma are enriched in oxidized mitochondrial DNA which is highly pro-inflammatory and pro-metastatic. J Cancer 2022; 13(4):1261-1271. doi:10.7150/jca.64170. Available from https://www.jcancer.org/v13p1261.htm

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Abstract

Graphic abstract

Background: Neutrophil extracellular traps (NETs) are net like extracellular structure formed by neutrophils in response to certain stimulation. It works as inflammatory regulator and metastasis promoter in cancer. Mitochondrial-(mt)DNA is a circular, mitochondria derived double strain molecule, which is involved in NETs formation. Its role in NETs induced inflammatory alteration in hepatocellular carcinoma (HCC) remained unexplored.

Method: We evaluated the mitochondrial reactive oxygen species (mitoROS) level in peripheral neutrophils from HCC patients and the oxidative level of mtDNA in derived NETs. The association between the NETs and oxidized mtDNA was assessed to reveal their relevance. A function assay was applied to uncover how the oxidation state of mtDNA directed the metastasis promoting inflammation state in HCC cells in a NETs protein dependent manner. Finally, using animal models, we explored the potential of a therapy strategy against NETs-drove metastasis by targeting the oxidized mtDNA with metformin.

Results: Neutrophils in HCC patients contained high level of mitoROS level, and formed NETs that were enriched in oxidized mtDNA in a mitoROS dependent manner. NETs and oxidized mtDNA were clinically relevant. Bound with NETs protein, oxidized mtDNA is more capable of triggering the metastasis-promoting inflammatory mediators in HepG2 cells. Targeting the oxidized mtDNA with metformin attenuated the metastasis-promoting inflammatory state and hereby undermine the metastasis capacity of HCC.

Conclusion: HCC is capable to stimulate NETs enriched in oxidized mtDNA, which are highly pro-inflammatory and pro-metastatic. Oxidized mtDNA in NETs may serve as a potential anti-metastatic target by metformin therapy.

Keywords: Neutrophil extracellular traps, hepatocellular carcinoma, oxidized mitochondrial DNA, metastasis