Journal: Redox Biology

Article Title: Integrated stress response-mediated metabolic reprogramming drives hepatic stellate cell activation and liver fibrosis via the noncanonical EIF3d-ATF4-S100P signaling pathway

doi: 10.1016/j.redox.2025.103905

Figure Lengend Snippet: EIF3d-ATF4 renders HSCs resistant to metabolic stress by augmenting S100P signaling. (A) Volcano plot comparing TGFβ1-treated HSCs with those treated with TGFβ1+siATF4 samples using DESeq-2. Benjamini-Hochberg method and Fisher's exact test were used. (B) GO enrichment analysis for each gene cluster was performed. Fisher's exact test and Benjamini-Hochberg method were used. (C) Expression of S100P in human LX-2 HSCs transfected with siEIF3d or siATF4 (n = 4). (D – E) Immunoblots suggested that EIF3d-ATF4 inhibition blocked S100P expression and ectopic expression of EIF3d-ATF4 stimulated S100P expression (n = 4). (F) Representative IHC images of S100P in liver sections from patients with fibrotic MASH (F0–F4) (n = 25). Black arrows suggest positive staining. Right: Correlation of relative S100P expression with METAVIR fibrosis scores. (G) RNA co-expression analysis of S100P and ATF4 in human liver tissues based on the GTEx database ( https://gtexportal.org ). (H) A scheme for the luciferase-reporter construct driven by S100P promoter. Luciferase activity results indicated that the transcriptional activities of S100P promoter were upregulated after TGFβ1 stimulation, while ATF4 deletion obviously reduced S100P transcriptional activity (n = 4). (I – J) Expression of fibrotic markers, S100P and metabolic enzymes after S100P deletion (n = 3). (K) Cell proliferation assay showed that ATF4 mediated increases in hHSCs proliferation activity are dependent on the activation of S100P (n = 6). (L – M) Flow cytometry analysis indicated that ectopic S100P expression markedly reduced ATF4 depletion induced HSCs apoptosis (annexin V-PI staining) (n = 4). Data are presented as means ± SEM; statistical analyses were determined by one-way ANOVA with SNK post hoc analysis (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: Antibodies used were EIF3d (Proteintech, 10219-1-AP), p -eIF2a (Cell Signaling, 3597L), ATF4 (Cell Signaling, 11815S), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling, 9251S), SAPK/JNK (Cell Signaling, 9252S), phospho-Erk (Thr202/Tyr204) (Cell Signaling, 4370S), phospho-p38 MAPK (Thr180/Tyr182) (Proteintech, 28796-1-AP), S100P (Proteintech, 11803-1-AP), NLRP3 (Proteintech, 19771-1-AP), COL1a1 (Cell Signaling, 72026S), SMAD3 (Cell Signaling, 9523T), p -SMAD3 (Cell Signaling, 9250T), S100P (Proteintech, 11803-1-AP), α-SMA (Cell Signaling, 1924S), GAPDH (Cell Signaling, 2118S).

Techniques: Expressing, Transfection, Western Blot, Inhibition, Staining, Luciferase, Construct, Activity Assay, Proliferation Assay, Activation Assay, Flow Cytometry

Journal: Redox Biology

Article Title: Integrated stress response-mediated metabolic reprogramming drives hepatic stellate cell activation and liver fibrosis via the noncanonical EIF3d-ATF4-S100P signaling pathway

doi: 10.1016/j.redox.2025.103905

Figure Lengend Snippet: Our novel small-molecule ERMT1 induces anti-fibrotic effects in vivo without toxicity. (A) Schematic of the high-throughput screening (HTS) pipeline used to identify small molecules that inhibit the ATF4-S100P pathway within the ISR. The screening employed a reporter cell line expressing ATF4-S100P promoter-driven luciferase, followed by downstream characterization. (B) Immunoblot analysis of LX-2 HSCs showing the effect of ERMT1 on the ATF4 pathway and fibrotic markers. Quantification of phosphorylated ATF4 and COL1a1 levels after 24 h of ERMT1 treatment (5–20 μg/ml) in the presence of TGFβ1 (10 nM), relative to the control (C). (D) Relative gene expression levels of α-SMA and COL1a1 in LX-2 HSCs following treatment with ERMT1 and TGFβ1. (E) Representative images of H&E, Masson's trichrome, and IHC staining for α-SMA in liver sections from mice treated with CCl 4 (0.5 μL/g, twice weekly for 8 weeks) and either ERMT1 or ISRIB (20 mg/kg, twice weekly, I.P.) (scale bars: 50 μm). Quantification of positive staining is shown in (H). (F) Serum levels of AST and ALT in CCl 4 -induced fibrotic mice. (G) mRNA expression of HSC markers in liver tissues from CCl 4 -treated mice, as indicated. (I) Representative liver tissue images of MCDD-induced liver fibrosis showing Masson's trichrome staining, α-SMA immunostaining, and Oil Red O (ORO) staining, with quantification of positive staining in (K) (scale bars: 50 μm). (J) Serum levels of AST and ALT in MCDD-induced fibrotic mice. (L) Relative gene expression levels of ATF4, S100P, and COL1a1 in liver tissues. Data are presented as means ± SEM. Statistical analyses were performed using one-way ANOVA followed by SNK post hoc analysis (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: Antibodies used were EIF3d (Proteintech, 10219-1-AP), p -eIF2a (Cell Signaling, 3597L), ATF4 (Cell Signaling, 11815S), phospho-SAPK/JNK (Thr183/Tyr185) (Cell Signaling, 9251S), SAPK/JNK (Cell Signaling, 9252S), phospho-Erk (Thr202/Tyr204) (Cell Signaling, 4370S), phospho-p38 MAPK (Thr180/Tyr182) (Proteintech, 28796-1-AP), S100P (Proteintech, 11803-1-AP), NLRP3 (Proteintech, 19771-1-AP), COL1a1 (Cell Signaling, 72026S), SMAD3 (Cell Signaling, 9523T), p -SMAD3 (Cell Signaling, 9250T), S100P (Proteintech, 11803-1-AP), α-SMA (Cell Signaling, 1924S), GAPDH (Cell Signaling, 2118S).

Techniques: In Vivo, High Throughput Screening Assay, Expressing, Luciferase, Western Blot, Control, Gene Expression, Immunohistochemistry, Staining, Immunostaining

Journal: BMC Cancer

Article Title: Integrative analysis of a novel signature incorporating metabolism and stemness-related genes for risk stratification and assessing clinical outcomes and therapeutic responses in lung adenocarcinoma

doi: 10.1186/s12885-025-13984-6

Figure Lengend Snippet: The expression patterns of prognostic risk genes and their correlation with OS in patients with LUAD. A The Kaplan-Meier plot depicts the relationship between the expression levels of S100P, GPX2, PRC2, ARNTL2, and RGS20 and OS in LUAD patients. B The expression patterns of S100P, GPX2, PRC2, ARNTL2, and RGS20 in LUAD and normal samples are presented based on data from the GEPIA database. C RT-qPCR analysis of S100P, GPX2, PRC2, ARNTL2, and RGS20 was conducted using matched clinical tissues ( n = 8). D Immunohistochemical analysis of S100P, GPX2, PRC2, ARNTL2, and RGS20 was performed on LUAD and normal tissue samples sourced from the Human Protein Atlas (HPA) database. HR, hazard ratio; OS, overall survival; LUAD, lung adenocarcinoma. Statistical significance is indicated as * P < 0.05 ; ** P < 0.01

Article Snippet: Furthermore, immunohistochemical analyses from the Human Protein Atlas (HPA) database demonstrated high staining intensity of S100P, GPX2, and PRC1 in LUAD tissues, contrasting with low intensity or absence of staining in normal tissues, while ARNTL2 and RGS20 did not exhibit significant differences (Fig. D).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining