Table_7_Regulation mechanisms of disulfidptosis-related genes in ankylosing spondylitis and inflammatory bowel disease.xlsx

IntroductionDisulfidptosis is a recently identified form of cell death that contributes to maintaining the internal environment balance of an organism. However, the molecular basis of disulfidptosis in ulcerative colitis (UC), ankylosing spondylitis (AS), and Crohn’s disease (CD) has not been thoroughly explored.
MethodsFirstly, the differentially expressed genes (DEGs) and disulfidptosis-associated genes (DAGs) were obtained through differential analysis between diseases (AS, CD, and UC) and control groups. After the disulfidptosis score was acquired using the single-sample gene set enrichment analysis (ssGSEA) algorithm, the DE-DAGs were screened by overlapping DAGs and DEGs of the three diseases. Next, the feature genes were selected through a combination of machine learning algorithms, receiver operating characteristic (ROC) curves, and expression analysis. Based on these feature genes, nomograms were created for AS, CD and UC. The co-feature genes were then identified by taking the intersections of the genes featured in all three diseases. Meanwhile, single-gene set enrichment analysis (GSEA) and the TF-mRNA-miRNA network were utilized to investigate the molecular mechanisms of the co-feature genes. To validate the expression differences of the co-feature genes between healthy controls and patients (AS and IBD), RT-PCR was performed. Lastly, mendelian randomization (MR) analysis was utilized to explore the causality between genetic variants of S100A12 with AS, UC and CD.
ResultsIn this study, 11 DE-DAGs were obtained. Functional enrichment analysis revealed their involvement in cytokine production and fatty acid biosynthesis. Latterly, AS/CD/UC -feature genes were derived, and they all had decent diagnostic performance. Through evaluation, the performance of the nomogram was decent for three diseases. Then, 2 co-feature genes (S100A12 and LILRA5) were obtained. The GSEA enrichment results indicated that the co-feature genes were mainly enriched in the cytokine-cytokine receptor interaction and drug metabolism cytochrome P450. As shown by functional experiments, there was a correlation between the mRNA expression of S100A12 with AS, UC and CD. Additionally, a causal connection between S100A12 and IBD was detected through MR analysis.
DiscussionIn this study, 2 co-feature genes (S100A12 and LILRA5) were screened, and their functions were investigated in AS, CD and UC, providing a basis for further research into diagnosis and treatment.

引言
二硫代死亡（disulfidptosis）是近年发现的一种细胞死亡形式，在维持机体内部环境稳态中发挥关键作用。然而，二硫代死亡在溃疡性结肠炎（UC）、强直性脊柱炎（AS）及克罗恩病（CD）中的分子基础尚未得到全面探索。

方法
首先，通过对疾病组（AS、CD、UC）与对照组进行差异表达分析，获取差异表达基因（DEGs）与二硫代死亡相关基因（DAGs）。采用单样本基因集富集分析（ssGSEA）算法计算二硫代死亡评分后，通过将三种疾病的DAGs与DEGs取交集，筛选出差异表达的二硫代死亡相关基因（DE-DAGs）。随后，结合机器学习算法、受试者工作特征（ROC）曲线及表达分析筛选特征基因。基于上述特征基因，分别为AS、CD及UC构建列线图模型。通过取三种疾病特征基因的交集，鉴定得到共同特征基因。同时，采用单基因集富集分析（GSEA）及转录因子（TF）-mRNA-微小RNA（miRNA）调控网络，探究共同特征基因的分子机制。为验证共同特征基因在健康对照与患者（AS及炎症性肠病（IBD））中的表达差异，本研究开展了逆转录聚合酶链式反应（RT-PCR）实验。最后，采用孟德尔随机化（MR）分析探讨S100A12的遗传变异与AS、UC及CD之间的因果关联。

结果
本研究共获得11个DE-DAGs。功能富集分析显示，这些基因参与细胞因子生成及脂肪酸生物合成过程。随后，分别得到AS、CD、UC的特征基因，且上述特征基因均具有良好的诊断效能。评估结果表明，针对三种疾病构建的列线图模型表现优异。最终鉴定得到2个共同特征基因（S100A12与LILRA5）。GSEA富集分析结果显示，共同特征基因主要富集于细胞因子-细胞因子受体相互作用及药物代谢细胞色素P450通路。功能实验结果表明，S100A12的mRNA表达水平与AS、UC及CD存在显著相关性。此外，孟德尔随机化分析证实S100A12与炎症性肠病（IBD）之间存在因果关联。

讨论
本研究筛选得到2个共同特征基因（S100A12与LILRA5），并探讨了其在AS、CD及UC中的功能，为上述疾病的诊断与治疗研究提供了理论依据。