N2 中性粒细胞刺激肝纤维化在威尔逊病中的作用。

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

机构信息

Institute of Translational Medicine, The Affiliated Hospital of Hangzhou Normal University, Hangzhou, Zhejiang, China.

Department of Hepatology, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Xinhua Hospital of Zhejiang Province, Hangzhou, Zhejiang, China.

出版信息

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

DOI:10.1016/j.jcmgh.2023.06.012

PMID:37406734

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10514429/

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.

摘要

背景与目的

威尔逊病是一种由铜转运体 ATP7B 基因突变引起的遗传性肝神经疾病。威尔逊病引起的肝脏疾病是青少年肝硬化的主要原因之一。目前的铜螯合剂和锌盐治疗方法改善了肝脏表现，但经常使神经症状恶化。在这项研究中，我们使用威尔逊病的斑马鱼/小鼠/细胞模型展示了中性粒细胞异质性的功能和机制。

方法

我们通过活细胞成像、运动追踪和分选细胞的转录分析研究了 atp7b 斑马鱼中的中性粒细胞反应。进行了实验以验证 Atp7b 小鼠中肝脏中性粒细胞的异质性。在 ATP7B 敲除的人肝癌 G2 细胞和分离的骨髓中性粒细胞中进行了体外实验，以揭示中性粒细胞异质性的机制。

结果

在 atp7b 斑马鱼中观察到中性粒细胞向肝脏的募集。中性粒细胞的药理学刺激加重了 atp7b 斑马鱼的肝脏和行为缺陷。对 atp7b 斑马鱼分选的肝脏中性粒细胞的转录分析揭示了一种独特的转录谱，特征为 N2 中性粒细胞。此外，在 ATP7B 敲除小鼠中也观察到肝脏 N2 中性粒细胞，并且药理学靶向转化生长因子 β1、DNA 甲基转移酶或信号转导和转录激活因子 3 减少了肝脏 N2 中性粒细胞，并改善了 ATP7B 敲除小鼠的肝功能和减轻了肝脏炎症和纤维化。转化生长因子 β1 对 Socs3 表达的表观遗传沉默有助于分离的骨髓中性粒细胞中 N2 中性粒细胞的极化。