Distinct histopathological phenotypes of severe alcoholic hepatitis suggest different mechanisms driving liver injury and failure
Jing Ma, … , Suthat Liangpunsakul, Bin Gao
Jing Ma, … , Suthat Liangpunsakul, Bin Gao
Published July 15, 2022
Citation Information: J Clin Invest. 2022;132(14):e157780. https://doi.org/10.1172/JCI157780.
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Research Article Gastroenterology

Distinct histopathological phenotypes of severe alcoholic hepatitis suggest different mechanisms driving liver injury and failure

Abstract

Intrahepatic neutrophil infiltration has been implicated in severe alcoholic hepatitis (SAH) pathogenesis; however, the mechanism underlying neutrophil-induced injury in SAH remains obscure. This translational study aims to describe the patterns of intrahepatic neutrophil infiltration and its involvement in SAH pathogenesis. Immunohistochemistry analyses of explanted livers identified two SAH phenotypes despite a similar clinical presentation, one with high intrahepatic neutrophils (Neuhi), but low levels of CD8+ T cells, and vice versa. RNA-Seq analyses demonstrated that neutrophil cytosolic factor 1 (NCF1), a key factor in controlling neutrophilic ROS production, was upregulated and correlated with hepatic inflammation and disease progression. To study specifically the mechanisms related to Neuhi in AH patients and liver injury, we used the mouse model of chronic-plus-binge ethanol feeding and found that myeloid-specific deletion of the Ncf1 gene abolished ethanol-induced hepatic inflammation and steatosis. RNA-Seq analysis and the data from experimental models revealed that neutrophilic NCF1-dependent ROS promoted alcoholic hepatitis (AH) by inhibiting AMP-activated protein kinase (a key regulator of lipid metabolism) and microRNA-223 (a key antiinflammatory and antifibrotic microRNA). In conclusion, two distinct histopathological phenotypes based on liver immune phenotyping are observed in SAH patients, suggesting a separate mechanism driving liver injury and/or failure in these patients.

Authors

Jing Ma, Adrien Guillot, Zhihong Yang, Bryan Mackowiak, Seonghwan Hwang, Ogyi Park, Brandon J. Peiffer, Ali Reza Ahmadi, Luma Melo, Praveen Kusumanchi, Nazmul Huda, Romil Saxena, Yong He, Yukun Guan, Dechun Feng, Pau Sancho-Bru, Mengwei Zang, Andrew MacGregor Cameron, Ramon Bataller, Frank Tacke, Zhaoli Sun, Suthat Liangpunsakul, Bin Gao

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Figure 7

Genetic deletion of the Ncf1 gene in neutrophils ameliorates chronic-plus-binge ethanol-induced liver ROS, inflammation, steatosis, and injury.

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Genetic deletion of the Ncf1 gene in neutrophils ameliorates chronic-plu...
WT and Ncf1Lyz–/– mice were fed an ethanol diet for 10 days, followed by gavage of ethanol, and were euthanized 9 hours later. (A) Serum ALT levels and circulating neutrophils were measured (WT, n = 14; KO, n = 12). (B) Liver tissues were subjected to H&E staining, TG measurement (WT, n = 12; KO, n = 12), and IHC staining (WT, n = 8; KO, n = 6) with antibodies against MDA, HNE, MPO, and F4/80. Representative images are shown on the left. Liver TG levels were quantified and are shown on the right. Quantification of the area positive for MDA or HNE staining is shown on the right. Scale bars: 200 μm (H&E row), 100 μm (additional rows). Quantification of number of MPO+ cells, percentage of F4/80+ area, and F4/80 clusters are on the right. Values in A and B represent mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001. (C and D) Heatmap illustrations of the expression profiles of genes involved in inflammation (C) and fibrosis (D) (WT, n = 6; KO, n = 6). P values are indicated. Statistical significance was assessed using 2-tailed Student’s t test for comparing 2 groups (A and B) and 1-way ANOVA followed by Tukey’s post hoc test for multiple groups (C and D).