Background: Type 2 diabetes mellitus (T2DM) is driven by progressive β-cell failure, wherein endoplasmic reticulum (ER) stress activates the Protein kinase R-Like ER kinase (PERK)-activating transcription factor 4 (ATF4) signaling cascade, promoting ferroptosis and accelerating β-cell dysfunction. This study investigated whether STC-1–derived exosomes (EXO) protect β-cells by modulating this pathway.

Methods: A T2DM mouse model was induced via high-fat diet (HFD) feeding and streptozotocin (STZ) administration. Exosomes were extracted from STC-1 cell culture supernatants, followed by both in vitro cellular experiments and in vivo animal studies. MIN6 cells cultured under high-glucose (HG) conditions were treated with graded concentrations of STC-1-EXO or the ferroptosis inhibitor Ferrostatin-1. Cell viability was quantified using cell Counting Kit-8 (CCK-8) assay. Western blotting (WB) assessed expression of ferroptosis markers acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4). Ferroptotic activity was evaluated by measuring intracellular total iron concentration, malondialdehyde (MDA) content, and glutathione (GSH) levels. In vivo, T2DM mice received STC-1-EXO treatment, followed by assessment of random blood glucose, glucose tolerance, insulin sensitivity, and pancreatic histopathology with concomitant protein expression analysis.

Results: In vitro, STC-1-EXO dose-dependently reversed HG-induced MIN6 cell viability loss (p < 0.05). Relative to the high-glucose control group, STC-1-EXO treatment significantly attenuated intracellular iron accumulation and MDA production, while restoring GSH levels and reducing lipid peroxidation (p < 0.05). WB results revealed that compared to the HG group, STC-1-EXO treatment significantly downregulated the expression of ER stress markers glucose-regulated protein 78 (GRP78), phosphorylated-PERK (p-PERK), phosphorylated eukaryotic translation initiation factor 2α (p-eIF2α), and ATF4, while simultaneously reducing the pro-ferroptotic protein ACSL4 expression and increasing the anti-ferroptotic protein GPX4 expression (p < 0.05). In vivo experiments showed that compared to untreated T2DM mice, STC-1-EXO-treated mice exhibited significantly decreased random blood glucose levels and markedly improved glucose tolerance and insulin sensitivity (p < 0.05). Hematoxylin and Eosin (H&E) staining revealed increased islet area and enhanced β-cell numbers in STC-1-EXO-treated mice. Furthermore, compared to untreated T2DM mice, STC-1-EXO treatment significantly reduced the expression of GRP78, p-PERK, p-eIF2α, and ATF4 in pancreatic tissue, decreased ACSL4 expression while increasing GPX4 expression, and resulted in significantly decreased MDA levels and elevated GSH levels in pancreatic tissue (p < 0.05).

Conclusion: STC-1-EXO effectively ameliorates ER stress and ferroptosis in pancreatic β-cells through inhibition of the PERK-ATF4 signaling pathway, thereby improving blood glucose levels and insulin secretory function in T2DM mice. This finding provides a novel theoretical foundation and potential therapeutic strategies for exosome-based diabetes treatment.

Background: The persistent expression of high-risk human papillomavirus (hrHPV) E6 oncoproteins is a critical determinant in driving and maintaining the malignant phenotype of cervical cancer, a pathogenic process where autophagy serves as a key regulatory mechanism. This study aimed to identify autophagy-related genes as potential biomarkers for prognostic evaluation in cervical cancer.

Methods: This study established an HPV 16 E6-induced C33a cervical cancer cell model, which was treated with the autophagy activator rapamycin or inhibitor 3-methyladenine to modulate autophagy. The expression of the autophagy-related gene C-X-C motif chemokine ligand 8 (CXCL8) was analyzed by reverse transcription-quantitative PCR (RT-qPCR), western blotting, and enzyme-linked immunosorbent assay. Cell viability, migration, invasion, and apoptosis were assessed using the Cell Counting Kit-8 assay, wound healing assay, Transwell assay, and Terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeling (TUNEL) assays, respectively. Furthermore, CXCL8 expression levels in tumor tissues from cervical cancer patients with favorable (n = 81) and unfavorable (n = 61) prognosis were examined by immunohistochemistry and RT-qPCR. Univariate and multivariate Cox proportional hazards regression analyses were conducted to identify independent risk factors influencing disease-free survival (DFS) in cervical cancer patients. Receiver operating characteristic (ROC) curve analysis was employed to evaluate the predictive value of CXCL8 for unfavorable prognosis risk.

Results: Mechanistic studies indicated the participation of CXCL8 in HPV 16 E6–induced malignant phenotypes. The autophagy activator rapamycin or CXCL8-neutralizing antibody could neutralize the oncogenic effects of HPV 16 E6. CXCL8 was highly expressed in tumor tissues with poor prognosis. It showed correlations with poor tumor differentiation, cervical infiltration depth ≥2/3, and lymph node metastasis, independent of clinical stage. CXCL8 was identified as an independent prognostic factor in cervical cancer [hazard ratio = 3.143, 95% confidence interval (CI): 1.519–6.507, p = 0.002] and was significantly correlated with inferior DFS (χ² = 34.905, p < 0.0001). Furthermore, a model combining CXCL8 with squamous cell carcinoma antigen and cytokeratin 19 fragment showed promising prognostic accuracy, achieving an area under the curve of 0.897 (95% CI: 0.835–0.942) with 90.16% sensitivity and 88.89% specificity.

Conclusion: The CXCL8 gene promotes cervical cancer progression by contributing to the regulation of autophagy mediated by HPV 16 E6. This functional role underpins its potential utility as a clinical prognostic biomarker.

Background: The Duffy Antigen Receptor for Chemokines (DARC/ACKR1) regulates inflammatory chemokine levels. The functional ACKR1 rs12075 G>A (p.Gly42Asp) polymorphism influences this activity and varies across populations, but studies on its association with human diseases such as coronary artery disease (CAD) are limited. The purpose of this study was to investigate the potential genetic link between this specific variation in the Duffy blood group gene ACKR1 and the risk of developing CAD within a Saudi Arabian population cohort.

Methods: A case-control study of 100 confirmed CAD patients and 100 matched healthy controls was conducted. Genomic DNA was extracted from peripheral blood and genotyped for the ACKR1 rs12075 variant using the Amplification Refractory Mutation System PCR (ARMS-PCR). Allelic and genotypic frequencies were compared using Chi-square tests, and associations with CAD risk were estimated via logistic regression, calculating odds ratios (ORs) with 95% confidence intervals (CI).

Results: The genotype distribution differed significantly between cases and controls (p = 0.0004). The AA genotype frequency was markedly higher in CAD patients (15%) versus controls (2%). In a recessive inheritance model, the AA genotype conferred a significantly increased risk of CAD (OR = 8.64, 95% CI: 1.92–38.90, p = 0.004). The GA genotype was also associated with elevated risk (OR = 2.06, 95% CI: 1.04–4.11, p = 0.037). Furthermore, the AA and GA genotypes showed significant associations with key CAD comorbidities, including hypertension, hyperlipidemia, diabetes mellitus, and a history of myocardial infarction (p < 0.05).

Conclusion: The ACKR1 rs12075 A allele, particularly in the homozygous state, is strongly associated with an increased risk of CAD in a Saudi Arabian cohort and is linked to a more severe clinical phenotype. These findings suggest that this genetic variant may serve as a potential biomarker for CAD susceptibility and severity in this population.

Background: Methotrexate (MTX) is the main rheumatoid arthritis (RA) therapy. However, the molecular pathways directly modulated by MTX and transcriptomic features that distinguish clinical responders from non-responders remain unclear. This study aimed to define MTX‑associated transcriptional programs and baseline synovial gene expression pathways associated with a clinical response to MTX-treated RA through cross‑tissue transcriptomic integration.

Methods: Peripheral blood gene expression profiles collected before and after MTX treatment were obtained from the Gene Expression Omnibus (GSE35455) database. Differential gene expression analysis was used to identify upregulated and downregulated genes, and functional enrichment analyses were conducted for each gene set. Clinical response–associated transcriptional signatures were independently evaluated on a dataset of baseline synovial tissue from MTX‑treated patients with RA (GSE45867). Gene set enrichment analysis was used to identify pathways associated with the MTX response.

Results: In the GSE35455 dataset, MTX treatment was associated with significant differential expression of 730 genes, which were predominantly downregulated. MTX-downregulated genes showed limited enrichment in classical Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways but demonstrated significant suppression of inflammatory signaling modules, including TNF-α signaling via NF-κB. In contrast, MTX-upregulated genes were enriched for metabolic stress responses, including Hypoxia and Glycolysis pathways, and innate immune response modules. Disease and phenotype enrichment analyses of the upregulated genes highlighted immune-related disease annotations and lymphoid tissue abnormalities. In synovial tissue (GSE45867), MTX responders exhibited strong baseline enrichment of interferon-stimulated gene programs, cytokine-regulated JAK-STAT signaling pathways, and immune activation signatures.

Conclusions: MTX induces broad immunomodulatory transcriptional effects characterized by the suppression of inflammatory signaling networks, and baseline activation of interferon- and cytokine-responsive programs in the synovial tissue. This cross-tissue integration provides mechanistic insights into MTX response heterogeneity in RA.