THEMIS attenuates MASH by suppressing disease-associated hepatocyte induction and hepatocyte senescence in mice
Xiaoxue Qiu, You Lu, Yuwei Tang, Linkang Zhou, Yu-tung Lee, Ziyi Meng, Zhimin Chen, Fnu Pradeepa, Lanuza A.P. Faccioli, Zhiping Hu, Alejandro Soto-Gutierrez, Siming Li, Jiandie D. Lin
Xiaoxue Qiu, You Lu, Yuwei Tang, Linkang Zhou, Yu-tung Lee, Ziyi Meng, Zhimin Chen, Fnu Pradeepa, Lanuza A.P. Faccioli, Zhiping Hu, Alejandro Soto-Gutierrez, Siming Li, Jiandie D. Lin
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Research Article Hepatology Metabolism

THEMIS attenuates MASH by suppressing disease-associated hepatocyte induction and hepatocyte senescence in mice

Abstract

Hepatocyte senescence is increasingly recognized as a pathogenic driver of metabolic dysfunction–associated steatohepatitis (MASH). Through single-nucleus transcriptomic profiling, we identified a discrete population of disease-associated hepatocytes (daHep) exhibiting enrichment for senescence markers in MASH livers. The emergence of senescent hepatocytes was associated with a marked induction of hepatic thymocyte selection associated (THEMIS) expression in both murine and human MASH. Genetic ablation of Themis, either globally or specifically in hepatocytes, resulted in significant expansion of daHep and senescent hepatocyte populations and exacerbated MASH pathology in mice. Single-nucleus transcriptomic analysis revealed a central role for THEMIS in shaping the cellular landscape of both parenchymal and nonparenchymal compartments within the MASH liver microenvironment. Conversely, adeno-associated virus–mediated overexpression of THEMIS suppressed hepatocyte senescence and attenuated diet-induced MASH. Mechanistic studies revealed that THEMIS deficiency promoted aberrant ERK phosphorylation and hepatocyte senescence. These findings establish THEMIS as a critical hepatoprotective factor that restrains hepatocyte senescence and mitigates metabolic liver disease progression.

Authors

Xiaoxue Qiu, You Lu, Yuwei Tang, Linkang Zhou, Yu-tung Lee, Ziyi Meng, Zhimin Chen, Fnu Pradeepa, Lanuza A.P. Faccioli, Zhiping Hu, Alejandro Soto-Gutierrez, Siming Li, Jiandie D. Lin

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Figure 1

Single-nucleus RNA-seq analysis of healthy and diet-induced MASH livers.

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Single-nucleus RNA-seq analysis of healthy and diet-induced MASH livers....
(A) UMAP clustering analysis of snRNA-seq dataset of 36,284 liver nuclei. (B) Hepatocyte subclusters in chow and MASH livers. (C) Relative proportion of chow and MASH hepatocytes for each subtype. (D) Heatmap illustrating the expression of marker genes for different hepatocyte subtypes. Hepatocytes in zones 1–3 were sorted along their coordinates in the first principal component (PC1) dimension. Each column represents average values binned from 500 neighboring cells. (E) Feature plots showing Nrg1 and Gdf15 expression in hepatocytes. (F) qPCR analysis of hepatic gene expression (chow, n = 4; MASH, n = 9). Statistical comparisons were conducted using 2-tailed unpaired Student’s t test. **P < 0.01, ***P < 0.001. (G) Immunoblotting analysis of total liver lysates. (H) Senescence gene score for different hepatocyte subtypes. Significance was estimated via 100 subsampling permutations (n = 30) using Wilcoxon’s rank-sum tests. Resulting P values were adjusted for multiple testing using the Benjamini-Hochberg FDR procedure. *P < 0.05, ***P < 0.001. (I) Visualization of hepatocyte senescence scores on UMAP coordinates. (J) Correlation between daHep and senescence gene signatures. (K) Violin plots of the normalized expression of Cdkn1a and Gdf15 in different hepatocyte subtypes. (L) RNA scope analysis of Nrg1 and Cdkn1a expression in chow and MASH liver sections. Arrows indicate colocalization. Scale bars: 50 μm.