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(A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and <t>SAA1).</t> (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.
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(A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and <t>SAA1).</t> (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.
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(A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and <t>SAA1).</t> (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.
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(A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and <t>SAA1).</t> (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.
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(A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and <t>SAA1).</t> (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.
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(A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and <t>SAA1).</t> (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.
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(A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and SAA1). (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.

Journal: bioRxiv

Article Title: Modulation of metabolic, inflammatory, fibrotic and cell death pathways by Resmetirom in metabolic dysfunction-associated steatohepatitis (MASH): A transcriptomic profiling study

doi: 10.1101/2025.11.26.690715

Figure Lengend Snippet: (A) Expression of thyroid hormone receptor-β ( THR-β ) and thyroid hormone receptor-α ( THR-α ) in human liver tissue across MASLD progression stages (GSE213621). (B) Experimental design: male C57BL/6J mice were fed a fructose, palmitate, and cholesterol (FPC) diet for 12 weeks. Therapeutic Resmetirom (Rem) (5 mg/kg/day) or vehicle (0.5% carboxymethyl cellulose-sodium + 0.4% Tween-80) was administered intraperitoneally from week 4 to week 12. (C) Body weight changes over the 12-week period. (D) Representative images and quantification of hepatic lipid accumulation by Nile Red, BODIPY, and hematoxylin & eosin (H&E) staining. Scale bar: 50 μm for Nile Red and BODIPY; Scale bar: 100 μm for H&E. (E–F) Serum biochemical analyses, including total cholesterol (TC), triglycerides (TG), high-density-lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). (G) Total cholesterol and triglyceride content in liver. (H) Immunofluorescence analysis of macrophage infiltration (CD68). Scale bar: 50 μm. (I-J) Serum levels of systemic inflammation markers (CRP and SAA1). (K–L) Sirius Red and immunofluorescence staining of α-smooth muscle actin (α-SMA) indicating fibrosis. Scale bar: 100 μm for Sirius Red; Scale bar: 50 μm for α-SMA. (M) TUNEL assay for hepatocyte apoptosis. Scale bar: 100 μm. (N) Principal component analysis (PCA) of hepatic transcriptome showing distinct clustering between vehicle- and Resmetirom-treated groups (n=4). (O) Volcano plot depicting differentially expressed genes (DEGs) between groups (p < 0.05, |log 2 FC| > 0.5). (P) Pathway enrichment analysis of DEGs highlighting key biological processes. (Q) Heatmap of representative DEGs associated with hepatic steatosis, inflammation, fibrosis, and cell death. Statistical significance: p < 0.05.

Article Snippet: To assess systemic inflammatory markers, serum concentrations of C-reactive protein (CRP; #KE10128, Proteintech) and serum amyloid A1 (SAA1; #CSB-EL020656MO, CUSABIO) were determined using specific ELISA kits following the manufacturers’ instructions.

Techniques: Expressing, Staining, Immunofluorescence, TUNEL Assay