Emerging evidence links reactive oxygen species modulator 1 (ROMO1), a key mitochondrial ROS regulator, to rheumatoid arthritis (RA) pathogenesis. However, its exact mechanism remains elusive given the conflicting evidence about its specific function. We used a four-level integrative framework combining multi-omics data and literature‑supported mechanistic inference. At the genetic level, Mendelian randomization (MR) was performed to explore potential causal relationships between ROMO1, IL2RA, HLA-DR, MIF, and RA risk, followed by differential expression analysis and machine learning-based feature selection to identify key mROS genes. The temporal expression dynamics of ROMO1 were assessed in RA progression. At the cellular and tissue levels, we integrated single-cell RNA sequencing and spatial transcriptomics to map cell-type-specific expression and synovial localization of ROMO1-related immune cells and pathways. Finally, our multi-omics findings were contextualized with literature-supported mechanistic inference. (1) MR results were consistent with a potential protective effect of ROMO1 on RA (OR = 0.52) and its potential regulation of risk factors IL2RA (OR = 0.46) and HLA-DR (OR = 0.40). Conversely, IL2RA (OR = 1.42), HLA-DR (OR = 1.88), and MIF (OR = 1.17) were positively associated with RA risk. Additionally, ROMO1 was identified as a top candidate diagnostic predictor with stage-specific dynamics: downregulated in the early but upregulated in the late/remission stages. (2) Single-cell RNA sequencing showed ROMO1's cell-specific expression in CD14+ HLA-DR+ CD74+ monocytes and CD4+ IL2RA+ T cells. Cell communication analysis further suggested that these cells may participate in MIF pathway regulation. Spatial transcriptomics subsequently identified that ROMO1-related cells localized to synovial pathological regions, with MIF pathway changes correlated with RA progression. (3) Finally, literature-supported mechanistic inference suggests that ROMO1 may modulate mROS levels to promote anti-inflammatory M2 macrophage polarization, which could theoretically contribute to reduced systemic inflammation and the alleviation of multi-organ decline in RA. This integrated multi-omics investigation, supported by literature-based mechanistic inference, suggests ROMO1 as a stage-dependent biomarker candidate and potential immune regulator in RA.
Characterized by renal inflammation and structural damage, lupus nephritis (LN) is a severe and often debilitating complication of systemic lupus erythematosus. The dynamic regulation of RNA processing, stability, and translation by N6-methyladenosine (m6A) modification has been implicated in the pathophysiology of LN. As an m6A reader, insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) plays a crucial role in the pathogenesis of various kidney diseases, yet its precise function in LN remains unclear. IGF2BP2 expression was measured in kidney specimens from LN patients and MRL/LPR mice. MRL/lpr mice were administered IGF2BP2 short hairpin RNA adeno-associated viral vectors, followed by evaluation of proteinuria, renal function, and histology. In vitro experiments employed lipopolysaccharide (LPS) stimulation of human kidney tubular epithelial cells (HK-2) to simulate LN inflammatory responses and further investigated the mechanism of IGF2BP2. Molecular interactions were validated via RNA immunoprecipitation and luciferase reporter assays. IGF2BP2 expression is significantly upregulated in LN and positively correlated with proinflammatory factor levels. Knockdown of IGF2BP2 significantly improved renal injury and alleviated renal inflammatory responses in MRL/LPR mice, as well as reduced LPS-induced inflammatory responses in HK-2 cells. Mechanistically, under LN pathological conditions, IGF2BP2 recognized m6A modifications on signal transducer and activator of transcription 1 (STAT1) mRNA and enhanced its stability, leading to the activation of proinflammatory signaling, exacerbation of renal inflammation, and ultimately renal injury. The m6A reader IGF2BP2 promotes proinflammatory signaling in LN by recognizing m6A-modified STAT1 mRNA. Targeting the IGF2BP2-STAT1 axis may represent a potential therapeutic strategy to ameliorate renal inflammatory responses in LN.
Tc17 cells and Tc17.1 cells contribute synovial inflammation during the development of rheumatoid arthritis (RA). However, the modulatory mechanisms of synovial Tc17 cells and Tc17.1 cells remain elusive. In the current study, we performed transcriptome profiling analysis on sorted live synovial Tc17 cells and Tc17.1 cells from a mouse model of collagen-induced arthritis (CIA). We found an array of differentially expressed genes (DEGs), which might be involved in the metabolism, cytokine signaling, and other biological aspects in synovial Tc17 and Tc17.1 cells. Importantly, we identified zinc finger protein 24 (ZNF24), a transcription factor, among the DEGs. ZNF24 knockdown impaired Tc17 differentiation and function in vitro, as evidenced by lower expression of IL-17A, RORγt, GM-CSF, IL-22, perforin, and granzyme B. ZNF24 knockdown resulted in similar effects on Tc17.1 differentiation and function in vitro. Furthermore, ZNF24 knockdown weakened the effects of Tc17 cells and Tc17.1 cells on inducing pro-inflammatory mediator expression in synovial fibroblasts. Adoptive transfer of Tc17.1 cells significantly exacerbated CIA development, whereas ZNF24-knockdown ameliorated the detrimental effect of Tc17.1 cells. Synovial fibroblasts up-regulated ZNF24 expression in Tc17 and Tc17.1 cells in a direct contact manner. The expression pattern and effect of ZNF24 were also observed in patients with RA and human Tc17 cells and Tc17.1 cells. Taken together, our study suggests that ZNF24 is a positive regulatory factor for Tc17 and Tc17.1 cells to exert pro-inflammatory effects in RA. We thus unveiled a novel mechanism by which the functions of synovial Tc17 cells and Tc17.1 cells are modulated.
Diffuse alveolar hemorrhage (DAH) is a lethal, female-biased complication of systemic lupus erythematosus (SLE). While adaptive immunity is a known driver, the role of innate lymphoid cells (ILCs) in DAH and the contribution of sex hormones to this process remain unclear. To characterize pulmonary ILC remodeling during DAH and evaluate whether androgen signaling modulates natural killer (NK) cell homeostasis and disease severity. A pristane-induced DAH model was established in C57BL/6J and Rag2 -/- mice. Pulmonary ILCs were profiled by single-cell transcriptomics and flow cytometry. Baseline sex differences were compared between healthy humans and mice. In vivo interventions included male castration, female testosterone propionate (TP) administration, and antibody-mediated NK cell depletion. DAH was accompanied by a marked reduction in pulmonary NK cell frequency in both C57BL/6J and Rag2 -/- mice, indicating an innate feature independent of adaptive lymphocytes. DAH-associated NK cells displayed an activated but constrained state, showing increased expression of effector-associated genes Prf1 and Ifng, together with the inhibitory receptor gene Lag3. At baseline, healthy human females and female mice exhibited lower NK cell frequencies than males. While male castration did not significantly alter DAH severity, TP administration in females restored pulmonary NK cell dominance and significantly attenuated disease severity. NK cell depletion prior to challenge led to a trend toward more severe hemorrhage (p = 0.084). Pulmonary NK cells are reduced in a lupus-associated DAH model and may participate in disease development. Androgen treatment restores pulmonary NK cell predominance and attenuates DAH severity, supporting further investigation of strategies aimed at restoring or preserving pulmonary NK cell homeostasis and function as potential therapeutic approaches for DAH.
BACKGROUND:Variants in the MT-TI gene, which encodes mitochondrial transfer RNA for isoleucine, have been associated with neuromuscular, cardiac, auditory, renal, and metabolic disorders, but their clinical interpretation remains difficult. OBJECTIVE:To integrate clinical, familial, heteroplasmy, and functional evidence across the reported MT-TI variant spectrum and clarify its implications for variant interpretation and diagnosis. METHODS:We conducted a narrative review of reported MT-TI variants, with detailed comparison of seven representative variants and synthesis of phenotypic, familial, tissue-specific heteroplasmy, and functional findings. RESULTS:Evidence was derived mainly from case reports and small pedigrees. Heteroplasmy differed markedly among blood, skeletal muscle, and myocardium, indicating that blood may not represent variant loads in energy-demanding tissues. Reported values generally reflected the lowest observed levels in affected individuals or family-specific boundaries rather than validated pathogenic cutoffs. Functional findings support a staged mechanism involving disturbed transfer RNA processing, structure, stability, or aminoacylation, followed by impaired mitochondrial protein synthesis and respiratory-chain dysfunction. Integrated mechanistic support was limited to a few variants, including m.4295A>G; evidence for most variants remained incomplete or indirect. CONCLUSION:Diagnosis requires tissue-informed heteroplasmy assessment integrated with phenotype, maternal family history, and functional evidence. Current treatment is supportive, and proposed reproductive and molecular strategies lack MT-TI-specific clinical-trial evidence.
BACKGROUND:The SOD1 gene encodes superoxide dismutase 1, an antioxidant enzyme in which pathogenic variants cause a subset of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Although the clinical and molecular consequences of SOD1 variants are well established, its locus-wide population genetic architecture has not been systematically characterized across global populations. OBJECTIVE:To characterize patterns of genetic diversity, population differentiation, and evolutionary constraint across the SOD1 locus in worldwide human populations. METHODS:We analyzed genomic variation spanning the SOD1 region in the five continental populations of the 1000 Genomes Project. Nucleotide diversity (π), fixation index (FST), and allele frequency distributions were estimated using sliding-window analyses. Bootstrap resampling was used to compare diversity within and outside the coding region. Recent positive selection was assessed using XP-nSL, and codon-based evolutionary analyses were performed using primate SOD1 orthologs. RESULTS:The coding region consistently exhibited a twofold to threefold reduction in nucleotide diversity relative to adjacent genomic sequences across all populations. Bootstrap analyses confirmed significantly lower diversity within the locus than in flanking regions (p < 0.05 in all populations). Population differentiation was low (mean FST ≈ 0.03) and showed no pronounced peaks within the coding interval, whereas allele frequency distributions were broadly similar across continental populations. XP-nSL analyses detected no evidence of population-specific recent selective sweeps. Codon-based analyses identified only 31 variable codons among 155 analyzed positions, six codons under negative selection, and no evidence of positive selection. CONCLUSION:The SOD1 locus exhibits reduced standing genetic variation, limited continental differentiation, and strong evolutionary conservation, consistent with sustained functional constraint. These findings provide a comprehensive population genetic framework for interpreting genetic variation at this medically important locus.
AIMS:There is no proven treatment to prevent the growth or rupture of abdominal aortic aneurysm (AAA). As an aneurysm enlarges over time, the risk of fatal aortic rupture increases. Metformin, a drug usually prescribed to treat Type 2 diabetes, has previously been associated with reduced AAA risk in observational studies. Our aim was to assess whether there was a causal association between metformin treatment and AAA using Mendelian randomisation (MR). METHODS:Logistic regression analysis was conducted in UK Biobank with 2972 AAA cases and 89,160 propensity-matched controls. In addition, two-sample MR analysis was performed, using a genetic proxy for metformin consisting of variants associated with both gene expression of seven metformin drug targets and decreased glycated haemoglobin (HbA1c) levels. Effect sizes for HbA1c were obtained from within UK Biobank, and for AAA risk from AAAgen, a multi-ancestry meta-GWAS analysis of 39,221 cases and 1,086,107 controls. RESULTS:We found evidence of a protective association between self-reported metformin treatment and reduced AAA risk in the observational analysis, OR 0.49 (95% CI: 0.41-0.59, p = 1.5 × 10-14). MR results support this finding, with an estimated decrease in AAA risk of 43%, OR = 0.57 (95% CI: 0.38-0.88, p = 0.010) per one standard deviation (sd) decrease in HbA1c via metformin gene targets, equivalent to the effect of a prescribed dose of metformin. This effect is specific to metformin target genes and was not seen using a general untargeted instrument. CONCLUSION:Our observational study found evidence that metformin use reduces the risk of developing AAA. The MR results support this finding, providing evidence that the association may be causal. Clinical trials are warranted to assess the efficacy of metformin to reduce the risk of aneurysm growth and rupture in people with AAA.
Interleukin 2 (IL-2) is a cytokine essential for the development and function of regulatory T cells, which are key players in maintaining immune tolerance by suppressing autoreactive lymphocytes. Patients with systemic lupus erythematosus (SLE) often exhibit reduced IL-2 production, resulting in impaired Treg function and a consequent breakdown of immune tolerance. Hematological abnormalities, mainly leukopenia and lymphopenia, are prominent features of SLE. This study aimed to explore the relationship between serum IL-2 concentrations and the altered blood cell populations observed in patients with SLE. We recruited 235 patients with SLE. Complete blood counts, including red blood cells, white blood cells, and platelets, were obtained, and IL-2 levels were quantified using the ultrasensitive single-molecule array (Simoa) technique. Multivariate linear and logistic regression analyses were performed to evaluate the associations between IL-2 concentrations and hematological parameters. Multivariate linear regression revealed significant inverse associations between the serum IL-2 concentration and the hemoglobin level, hematocrit, mean corpuscular hemoglobin concentration, and total leukocyte, neutrophil, lymphocyte, and basophil counts. Similarly, after adjustment for covariates, serum IL-2 was associated with increased odds of anemia, leukopenia, and lymphopenia but not neutropenia or thrombocytopenia. In conclusion, circulating IL-2 levels are independently associated with specific hematological abnormalities, including anemia, leukopenia, and lymphopenia, in patients with SLE. These findings highlight the potential role of IL-2 and regulatory T cell dysfunction in the hematologic manifestations of SLE.
The advancement of hepatocellular carcinoma (HCC) is associated with metabolic reprogramming and immune escape. Compound kushen injection (CKI) is a cancer therapeutic agent with a long history. O-GlcNAcylation is reported to regulate tumor progression. We aimed to investigate how CKI affects HCC cell glycolysis and immune evasion and then clarify the underlying O-GlcNAcylation regulatory mechanism. HCC cell lines and subcutaneous transplantation mice were treated with CKI, and glycolysis and immune evasion were analyzed. The O-GlcNAcylation was analyzed using immunoprecipitation and western blotting. The results showed that CKI inhibited the viability, glycolysis, and immune escape of HCC cells, as well as hindered tumor growth, glycolysis, and immune evasion in vivo. Induction of glycolysis caused by hypoxia reduced the effect of CKI on immune evasion. Besides, CKI reduced the O-GlcNAc levels and OGT transcription activity. Overexpression of OGT counteracted the inhibition of immune evasion caused by CKI. Additionally, OGT catalyzed the O-GlcNAcylation of PD-L1 and enhanced its stability. In conclusion, CKI suppresses immune evasion of HCC cells in association with glycolysis suppression, which is related to the reduction of PGT-mediated PD-L1 O-GlcNAcylation. These findings suggest that CKI possesses an anti-HCC effect, and targeting PD-L1 O-GlcNAcylation may improve its therapeutic efficacy on HCC.
Increasing evidence shows that CD8+ T cells are the pathogenic mediators of tissue injury in systemic lupus erythematosus (SLE), sustaining the chronic inflammation through the accumulation of long-lived cytotoxic memory populations. However, the transcriptional mechanisms that prevent the aberrant differentiation of pathogenic CD8+ T cells remain poorly understood. Here, the transcription factor E4BP4 (NFIL3) was identified as a critical restraint of cytotoxic effector-memory CD8+ T cells in lupus. E4BP4 expression was reduced in CD8+ T cells from SLE patients and inversely correlated with disease activity. Using a lupus-like disease model, we found that E4BP4 deficiency accelerated disease progression, resulting in heightened autoantibody production, immune complex deposition, and renal pathology. This phenotype was associated with the systemic accumulation of cytotoxic effector-memory CD8+ T cells. Depletion of CD8+ T cells significantly ameliorated the disease phenotype, confirming the functional contribution of CD8+ T cells to lupus-like immunopathology. Competitive adoptive transfer experiments revealed that E4BP4 functions cell-intrinsically to limit the cytotoxicity and proliferation of CD8+ T cells in autoimmunity. Beyond autoimmunity, E4BP4 deficiency also resulted in an exuberant CD8+ effector-memory T cell response to Listeria monocytogenes infection, indicating a broader role for E4BP4 in limiting CD8+ T cell effector-memory responses. Collectively, these findings establish E4BP4 as a transcriptional checkpoint that restricts pathogenic CD8+ effector-memory T cell responses to maintain immune homeostasis in autoimmunity and infection.
Myasthenia gravis (MG) is a complex autoimmune neuromuscular disorder, and the role of lipid metabolism dysregulation in MG pathogenesis remains unclear. This study aimed to investigate the molecular mechanisms of lipid metabolism regulation in myasthenia gravis development by integrating single-cell and bulk transcriptomic data. This study analyzed bulk RNA sequencing data from the GSE85452 dataset (13 MG patients and 12 healthy controls) and single-cell RNA sequencing data from the GSE227835 dataset (10 MG patients and 10 healthy controls). Differential expression analysis was performed, and weighted gene co-expression network analysis (WGCNA) was conducted to identify disease-related modules. Key genes were screened through the intersection of differentially expressed genes (DEGs), WGCNA hub genes, and lipid metabolism-related genes. Functional enrichment analysis, protein‒protein interaction (PPI) network construction, immune infiltration analysis, and regulatory network analysis were performed. Single-cell analysis was used to characterize cellular heterogeneity and intercellular communication features. A nomogram prediction model was constructed and internally validated using leave-one-out cross-validation (LOOCV). Potential therapeutic compounds were identified through drug prediction and molecular docking analysis. Furthermore, key genes were validated by RT-qPCR in an independent cohort of 5 MG patients and 5 healthy controls. A total of 823 DEGs and 13 co-expression modules were identified, of which 4 modules were significantly associated with MG. Twenty-one candidate genes were screened, and 2 key genes (IRS2 and ALDH2) were ultimately determined. IRS2 was significantly downregulated while ALDH2 was significantly upregulated in MG patients. The nomogram model based on key genes demonstrated excellent predictive performance (AUC = 0.897). Immune infiltration analysis showed increased regulatory T cells (Tregs) and decreased CD4+ memory activated T cells in MG patients. Single-cell analysis identified 10 major cell types. Cell‒cell communication analysis revealed dense interactions among CD4+ T cells, B cells, and CD14+ monocytes. Drug prediction identified metformin and cyclophosphamide as potential therapeutic candidates, and molecular docking confirmed favorable binding affinities. This integrative study generated the hypothesis that lipid metabolism dysregulation, potentially mediated by IRS2 and ALDH2, may contribute to immune dysfunction in MG pathogenesis. These findings provide preliminary insights into the molecular mechanisms underlying MG development and suggest potential diagnostic biomarkers and therapeutic targets. However, these results are hypothesis-generating, and clinical translation is contingent upon rigorous mechanistic validation through functional experiments, animal models, and larger multicenter clinical cohorts.
While interferon regulatory factor 1 (IRF1) has been implicated in reactive oxygen species (ROS)-dependent neutrophil extracellular trap (NET) release during acute lung injury, its regulatory role in mitochondrial dysfunction-driven NETosis specific to pediatric pneumonia remains unexplored. Neutrophils were isolated from the bronchoalveolar lavage fluid (BALF) of pediatric pneumonia patients and the bone marrow of mice. IRF1 expression was quantified via qRT-PCR and Western blotting. Mitochondrial ROS (mtROS) and total ROS were measured by flow cytometry. Mitochondrial dysfunction was assessed by ATP quantification. NETosis was evaluated through immunofluorescence staining and ELISA quantification of neutrophil elastase (NE), myeloperoxidase-DNA (MPO-DNA), and citrullinated histone H3 (Cit-H3) levels. For in vivo studies, pneumonia-related lung injury was induced by intratracheal LPS instillation in mice, with pathological severity graded by H&E staining, pulmonary edema quantified via the wet/dry weight ratio, and inflammation assessed by the BALF protein concentration. Mechanistically, the IRF1/MMP9 interaction was predicted by bioinformatics (STRING database) and validated by co-immunoprecipitation (Co-IP) and immunofluorescence staining, while MMP9 overexpression was achieved via lentivirus transduction to delineate pathway regulation. IRF1 was significantly upregulated in BALF neutrophils from patients and correlated with elevated ROS production and mitochondrial dysfunction, as well as NETs release. IRF1 knockdown attenuated ROS-driven NETosis in vitro. Matrix metalloproteinase 9 (MMP9) was predicted to interact with IRF1, and MMP9 overexpression effectively reversed the beneficial effects of IRF1 deficiency on ROS release, mitochondrial dysfunction, neutrophil apoptosis, and NETosis. Consistently, in mouse models, MMP9 overexpression abolished the protective effects of IRF1 deficiency, exacerbating acute lung injury and restoring NETs levels in BALF.
OBJECTIVE:To investigate the effects of Platycodin D (PD) on the proliferation, migration, and angiogenesis of EA.hy926 endothelial cells stimulated by supernatant from a rheumatoid arthritis (RA) synovial cell model (MH7A), this study aims to preliminarily explore its potential mechanisms. METHODS:The TNF-α-stimulated MH7A cells were utilized as a model for rheumatoid arthritis (RA) cells. The supernatant from these cells was collected and designated as conditioned medium (CM), which was then used to stimulate EA.hy926 cells, thereby establishing an RA endothelial cell model; EA.hy926 cells were transduced with lentivirus to overexpress or knock down CD146; The experimental groups included: a normal control group, a model group (optimal CM stimulation group), and PD treatment groups (PD at concentrations of 1.25, 2.5, and 5 mg/L); sh-CD146 + CM, sh-NC + CM, sh-NC, lenti-CD146 + CM, lenti-CD146-NC. The rescue experiment included the following groups: CM, CM + PD group, CM + lenti-CD146 group, and CM + lenti-CD146 + PD treatment group. Cell proliferation was assessed using the CCK-8 assay; cell migration was evaluated through the wound-healing assay; angiogenic capability was determined via tube formation assays; and CD146 expression levels were measured by Western blotting. RESULTS:Compared to the normal group, CM stimulation significantly enhanced endothelial cell proliferation (P < 0.01). However, after 24 hours of PD treatment, a notable decrease in cell proliferation was observed (P < 0.05). Additionally, CM stimulation improved cell migration ability relative to the normal group; this enhancement was significantly diminished following PD treatment (P < 0.01). Furthermore, tube formation capability was markedly increased with CM stimulation compared to the normal group, but showed significant inhibition after 24 hours of PD treatment (P < 0.0001). Moreover, CD146 expression levels were significantly elevated in the model group when compared to the normal group and subsequently decreased following PD treatment (P < 0.01), and in a dose-dependent manner. In the rescue experiment, compared with the PD treatment group (CM + PD 5 mg/L), the proliferation, migration, and tube formation abilities of cells in the CD146 overexpression + PD treatment group (CM + lenti-CD146 + PD 5 mg/L) were significantly increased (P < 0.0001, P < 0.05, P < 0.001). Interestingly, when compared with the overexpression group (CM + lenti-CD146), these abilities in the overexpression+ PD treatment group (CM + lenti-CD146 + PD 5 mg/L) remained significantly suppressed (P < 0.0001, P < 0.0001, P < 0.05). CONCLUSION:In this study, we further found that CD146 exhibits significant pro-angiogenic, pro-migratory, and pro-proliferative effects in EA.hy926 cells. Meanwhile, PD was shown to significantly inhibit the angiogenesis, proliferation, and migration of EA.hy926 cells in a dose-dependent manner, and downregulate CD146 expression in a dose-dependent manner. CD146 overexpression partially attenuated PD's anti-angiogenic effect, but PD still exhibited strong anti-angiogenic activity in CD146-overexpressing cells, suggesting that PD acts partly through CD146 regulation and possibly other pathways, reflecting its multifaceted nature. This study is the first to link PD's anti-angiogenic effect to CD146, revealing a promising therapeutic direction for RA.
Objective This study aims to screen for genetic variants associated with premature ovarian insufficiency (POI) in a Chinese Miao pedigree. Methods The proband underwent whole exome sequencing (WES), and the extracted data were subjected to bioinformatics analysis using the Rare Disease Data Center (RDDC) splicing tool to identify potential genetic causes in the lineage. Sanger sequencing was employed to confirm the variants in the family, and minigene assays were used to analyze how the FIGLA variant affects pre-mRNA splicing. Results A novel heterozygous intronic and POI-associated variant in the FIGLA gene (c.385-9G>A) was identified. Splice prediction analysis showed that this variant may disrupt splicing patterns. This result was supported by the outcome of minigene assays, which revealed that the variant led to aberrant splicing of FIGLA introns. As a result, retention of a 7-bp intronic sequence from intron 2 was found in the mature mRNA. Conclusion The FIGLA variant (c.385-9G>A) disrupts mRNA splicing in cells and may contribute to POI in this pedigree. Therefore, our results expand the variant spectrum of the FIGLA gene, provide valuable insights into the pathogenesis of POI for genetic counseling, and may assist clinicians in the early diagnosis of women with infertility.
Hashimoto's thyroiditis (HT) is a common autoimmune thyroid disorder with a complex genetic background. Interleukin-6 (IL-6) has been implicated in its pathogenesis, but the genetic mechanisms remain unclear. Two-sample Mendelian randomization (MR) using expression quantitative trait loci (eQTL) and gene set enrichment analysis (GWAS) data identified IL‑6-HT genes. In vivo validation in nonobese diabetic (NOD) mice involved siRNA knockdown, histopathology HE/Immunohistochemistry (HE/IHC), and enzyme-linked immunosorbent assay (ELISA). GSEA and molecular docking explored mechanisms. Western blot in THP‑1 cells assessed Nuclear factor kappa-light-chain-enhancer of activated B cells (NF‑κB) pathway proteins (pIKKβ, IKKβ, pTAK1, TAK1, pp65, p65) and associations with SLC45A3/CCDC77 at 2, 24, 48 h (β‑actin control). MR analysis identified CCDC77 and SLC45A3 as key genes associated with both IL-6 and HT. In vivo experiments showed that knockdown of CCDC77 exacerbated thyroid damage, increased serum anti-thyroperoxidase antibody (TPOAb), anti-thyroglobulin antibody (TGAb), and IL-6 levels, while reduced free triiodothyronine (FT3) and free thyroxine (FT4). Conversely, SLC45A3 knockdown alleviated thyroiditis, reduced antibody levels, and improved thyroid function. GSEA Western blot analysis further revealed that both CCDC77 and SLC45A3 were involved in regulating the activation of NF‑κB signaling pathway. Our study demonstrated that SLC45A3 and CCDC77 were involved in modulating HT progression through NF‑κB-IL-6 axis, providing novel therapeutic strategies for clinical applications.
BACKGROUND:Preeclampsia (PE) is a complex pregnancy disorder associated with early placental hypoxia, oxidative stress, and impaired angiogenic signaling. Melatonin and soluble tumor necrosis factor like weak inducer of apoptosis (sTWEAK) contribute to antioxidant and vascular pathways, whereas their receptors, melatonin receptor 1A (MTNR1A), melatonin receptor 1B (MTNR1B), and fibroblast growth factor-inducible 14 (Fn14), may be subject to epigenetic regulation. This study assessed serum melatonin and sTWEAK levels in parallel with promoter methylation of MTNR1A, MTNR1B, and Fn14 in early gestation. METHODS:A mixed design cohort was recruited between 13 and 20 weeks of gestation. A total of 198 pregnant women were categorized as pregnant control, high risk, or preeclamptic at enrollment time. Serum melatonin and sTWEAK were measured by enzyme-linked immunosorbent assay (ELISA). Promoter methylation of MTNR1A, MTNR1B, and Fn14 was assessed using methylation-specific PCR with semi quantitative densitometry. RESULTS:Serum sTWEAK levels were significantly lower in high-risk and preeclamptic women, whereas melatonin levels showed a downward trend without reaching statistical significance. Promoter methylation of MTNR1A and Fn14 was elevated in both high-risk and preeclamptic groups, whereas MTNR1B showed no notable differences. Multivariate analysis revealed that lower sTWEAK (OR 0.837; 95% confidence interval [CI] 0.782-0.896; p < 0.001) and higher Fn14 methylation (OR 0.935; 95% CI 0.882-0.992; p = 0.027) were independently associated with hypertensive outcomes. Additionally, higher Fn14 methylation in early pregnancy was observed in high-risk women who later developed hypertensive disorders. CONCLUSION:Early pregnancy hypermethylation of MTNR1A and Fn14, but not MTNR1B, was observed in high-risk and PE women and co-occurred with a significantly reduced sTWEAK level and non-significantly reduced serum melatonin, suggesting epigenetic modulation of antioxidant and angiogenic pathways in women at risk for PE. These findings establish associations rather than causation and require validation using quantitative assays in multicenter cohorts before clinical translation.
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease with complex molecular mechanisms. Although transcriptomic studies have revealed prominent interferon signatures in SLE, robust prioritization of candidate genes across independent cohorts remains challenging. In this study, we performed a multi-cohort transcriptomic analysis using publicly available GEO datasets. Differential expression analysis was conducted independently within each cohort to minimize cross-study confounding. Machine learning models, including LASSO, support vector machine, and random forest, were applied for feature prioritization in a designated training cohort, followed by independent validation in separate datasets. Model interpretability was assessed using SHAP analysis. Immune cell composition was estimated descriptively using CIBERSORT. In addition, molecular docking and molecular dynamics simulations were performed as exploratory in silico analyses to evaluate potential protein-compound interactions. A set of consistently dysregulated genes across cohorts was identified, many of which are associated with interferon signaling. Among these, RSAD2 showed robust prioritization across multiple machine learning models. The predictive performance of the models was stable in independent validation datasets. SHAP analysis highlighted the contribution of interferon-stimulated genes to model predictions. Immune deconvolution suggested altered immune cell composition in SLE samples, consistent with previously reported immune activation patterns. Exploratory in silico analyses suggested a potential interaction between artemisinin and RSAD2. This study provides a robust, multi-cohort computational framework for prioritizing candidate genes associated with SLE. The findings highlight interferon-associated transcriptional features as reproducible molecular signatures of SLE and generate testable hypotheses for future experimental and clinical investigation.
IgA vasculitis (IgAV) primarily affects small vessels, but rare cases with necrotizing arteritis (NA) raise questions about overlap with polyarteritis nodosa (PAN). To characterize IgAV with necrotizing arteritis (IgAV-NA) and compare its phenotype with classical IgAV and PAN. We performed a multicenter retrospective study combined with a systematic literature review (1990-2025). Patients fulfilled EULAR/PRINTO/PRES IgAV criteria, had pathological or imaging evidence of NA in small or medium arteries, and were ANCA-negative. Thirty patients were included (7 from databases, 23 from the literature). NA was confirmed by biopsy (n = 16) or vascular imaging (n = 14). Clinical features, treatments, remission, and mortality were compared with 257 adult IgAV and 196 PAN patients. Median age was 54.5 years. IgAV-NA was characterized by severe manifestations, including gastrointestinal bleeding, perforation, surgical abdomen, neuropathy, pancreatitis, and livedo. Compared with classical IgAV, IgAV-NA showed significantly higher rates of multi-organ involvement and mortality. Compared with PAN, IgAV-NA shared vascular complications but had less fever and neuropathy. Despite arterial involvement, patients did not fulfil PAN criteria. IgAV-NA represents a rare, severe IgAV phenotype with life-threatening complications rather than an IgAV-PAN overlap. Severe or atypical IgAV presentations should prompt vascular imaging and intensified immunosuppression.
To systematically examine global research trends in the association between ferroptosis and autoimmune diseases from 2018 to 2025, thereby informing future mechanistic and translational investigations in this field. On March 17, 2026, relevant publications were obtained from the Web of Science Core Collection and PubMed databases. This included articles and reviews written in English and published between 2018 and 2025. Bibliometric visualization was conducted using CiteSpace 6.4.R1, VOSviewer 1.6.20, and Scimago Graphica to map countries, authors, institutions, keywords, journals, and references. Data processing and descriptive statistical analysis were performed using Microsoft Excel 2019. Seven hundred and nineteen publications from 27 countries were included in the analysis. The annual number of publications showed a consistent upward trend, with China contributing the largest share (487 publications). High-frequency keywords included rheumatoid arthritis, ulcerative colitis, inflammatory bowel disease, oxidative stress, and lipid peroxidation, indicating research hotspots in this field. Research on the association between ferroptosis and autoimmune diseases has expanded substantially over the past 8 years. However, the field remains in a relatively early stage of development. These findings suggest that the field is receiving increasing scholarly attention and may provide useful references for future biomarker discovery and novel therapeutic strategy development.
Monocytes are major contributors to systemic lupus erythematosus (SLE) pathogenesis, modulating B- and T-cell autoreactivity through inflammatory cytokine secretion and disrupted immune clearance. Under prolonged inflammatory pressure, monocytes develop a pathogenic “exhausted” immune memory state defined by paradoxical proinflammatory and immunosuppressive gene expression and diminished immune effector functions. To determine whether chronic inflammation in SLE elicits a similar monocyte exhaustion phenotype, we analyzed bone marrow and splenic reservoir monocytes from lupus-prone MRL and MRL/lpr (LPR) mice. Monocytes from LPR mice exhibit chronic immune exhaustion, including reduced chemokine receptor CX3CR1 cell surface levels, heightened sensitivity to endotoxin stress, and altered expression of T cell regulatory molecules PD-L1, CD200R, ICOS-L, and CD86. Exhaustion severity correlated with SLE progression, although the development of monocyte exhaustion preceded the onset of disease symptoms. These features were largely recapitulated in monocytes from patients with SLE, demonstrating that monocyte exhaustion is a generalizable feature of SLE pathogenesis.
