Stimulation of Liver Fibrosis by N2 Neutrophils in Wilson's Disease

Xiaoxiao Mi; Yu Song; Chaohua Deng; Jian Yan; Zhihui Li; Yingniang Li; Jun Zheng; Wenjun Yang; Ling Gong; Junping Shi

doi:10.1016/j.jcmgh.2023.06.012

Stimulation of Liver Fibrosis by N2 Neutrophils in Wilson's Disease

Cell Mol Gastroenterol Hepatol. 2023;16(5):657-684. doi: 10.1016/j.jcmgh.2023.06.012. Epub 2023 Jul 3.

Authors

Xiaoxiao Mi¹, Yu Song², Chaohua Deng³, Jian Yan¹, Zhihui Li⁴, Yingniang Li⁵, Jun Zheng⁶, Wenjun Yang⁶, Ling Gong³, Junping Shi⁷

Affiliations

¹ Institute of Translational Medicine, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China.
² Department of Hepatology, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Xinhua Hospital of Zhejiang Province, Hangzhou, Zhejiang, China.
³ Department of Infectious Diseases and Hepatology, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China.
⁴ Clinical Research Center, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
⁵ School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
⁶ Department of Pathology, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China.
⁷ Institute of Translational Medicine, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China; Department of Infectious Diseases and Hepatology, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China; Institute of Hepatology and Metabolic Diseases, Hangzhou Normal University, Zhejiang, China. Electronic address: 20131004@hznu.edu.cn.

Abstract

Background & aims: Wilson's disease is an inherited hepatoneurologic disorder caused by mutations in the copper transporter ATP7B. Liver disease from Wilson's disease is one leading cause of cirrhosis in adolescents. Current copper chelators and zinc salt treatments improve hepatic presentations but frequently worsen neurologic symptoms. In this study, we showed the function and machinery of neutrophil heterogeneity using a zebrafish/murine/cellular model of Wilson's disease.

Methods: We investigated the neutrophil response in atp7b^-/- zebrafish by live imaging, movement tracking, and transcriptional analysis in sorted cells. Experiments were conducted to validate liver neutrophil heterogeneity in Atp7b^-/- mice. In vitro experiments were performed in ATP7B-knockout human hepatocellular carcinomas G2 cells and isolated bone marrow neutrophils to reveal the mechanism of neutrophil heterogeneity.

Results: Recruitment of neutrophils into the liver is observed in atp7b^-/- zebrafish. Pharmacologic stimulation of neutrophils aggravates liver and behavior defects in atp7b^-/- zebrafish. Transcriptional analysis in sorted liver neutrophils from atp7b^-/- zebrafish reveals a distinct transcriptional profile characteristic of N2 neutrophils. Furthermore, liver N2 neutrophils also were observed in ATP7B-knockout mice, and pharmacologically targeted transforming growth factor β1, DNA methyltransferase, or signal transducer and activator of transcription 3 reduces liver N2 neutrophils and improves liver function and alleviates liver inflammation and fibrosis in ATP7B-knockout mice. Epigenetic silencing of Socs3 expression by transforming growth factor β1 contributes to N2-neutrophil polarization in isolated bone marrow neutrophils.

Conclusions: Our findings provide a novel prospect that pharmacologic modulation of N2-neutrophil activity should be explored as an alternative therapeutic to improve liver function in Wilson's disease.

Keywords: Liver Fibrosis; N2 Neutrophils; Neutrophil Heterogeneity; Wilson’s Disease.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adolescent
Animals
Copper / metabolism
Hepatolenticular Degeneration* / genetics
Hepatolenticular Degeneration* / metabolism
Humans
Liver Cirrhosis / pathology
Liver Neoplasms* / pathology
Mice
Mice, Knockout
Neutrophils / metabolism
Transforming Growth Factor beta1
Zebrafish / metabolism

Substances

Transforming Growth Factor beta1
Copper