G-CSF increases calprotectin expression, liver damage and neuroinflammation in a murine model of alcohol-induced ACLF

Martí Ortega-Ribera; Yuan Zhuang; Veronika Brezani; Prashanth Thevkar Nagesh; Radhika S Joshi; Mrigya Babuta; Yanbo Wang; Gyongyi Szabo

doi:10.3389/fcell.2024.1347395

G-CSF increases calprotectin expression, liver damage and neuroinflammation in a murine model of alcohol-induced ACLF

Front Cell Dev Biol. 2024 Feb 14:12:1347395. doi: 10.3389/fcell.2024.1347395. eCollection 2024.

Authors

Martí Ortega-Ribera¹, Yuan Zhuang¹, Veronika Brezani¹, Prashanth Thevkar Nagesh¹, Radhika S Joshi¹, Mrigya Babuta¹, Yanbo Wang¹, Gyongyi Szabo^{1

2}

Affiliations

¹ Department of Medicine, Division of Gastroenterology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, United States.
² Broad Institute, Cambridge, MA, United States.

Abstract

Background and aims: Granulocyte colony-stimulating factor (G-CSF) has been proposed as a therapeutic option for patients with ACLF, however clinical outcomes are controversial. We aimed at dissecting the role of G-CSF in an alcohol-induced murine model of ACLF. Methods: ACLF was triggered by a single alcohol binge (5 g/kg) in a bile duct ligation (BDL) liver fibrosis model. A subgroup of mice received two G-CSF (200 μg/kg) or vehicle injections prior to acute decompensation with alcohol. Liver, blood and brain tissues were assessed. Results: Alcohol binge administered to BDL-fibrotic mice resulted in features of ACLF indicated by a significant increase in liver damage and systemic inflammation compared to BDL alone. G-CSF treatment in ACLF mice induced an increase in liver regeneration and neutrophil infiltration in the liver compared to vehicle-treated ACLF mice. Moreover, liver-infiltrating neutrophils in G-CSF-treated mice exhibited an activated phenotype indicated by increased expression of CXC motif chemokine receptor 2, leukotriene B4 receptor 1, and calprotectin. In the liver, G-CSF triggered increased oxidative stress, type I interferon response, extracellular matrix remodeling and inflammasome activation. Circulating IL-1β was also increased after G-CSF treatment. In the cerebellum, G-CSF increased neutrophil infiltration and S100a8/9 expression, induced microglia proliferation and reactive astrocytes, which was accompanied by oxidative stress, and inflammasome activation compared to vehicle-treated ACLF mice. Conclusion: In our novel ACLF model triggered by alcohol binge that mimics ACLF pathophysiology, neutrophil infiltration and S100a8/9 expression in the liver and brain indicate increased tissue damage, accompanied by oxidative stress and inflammasome activation after G-CSF treatment.

Keywords: BDL; S100a8/S100a9; acute-on-chronic liver failure; cerebellum; encephalopathy; ethanol; inflammasome; neutrophil.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study was supported by NIH grants R01 AA017729, R01 AA020744, and R01 AA011576 (to GS).