Anoikis-Related Gene EZR is a Potential Factor Associated with Neutrophil Extracellular Traps(NETs) in Ankylosing Spondylitis with Hip Arthropathy

Abstract

Abstract Background: Ankylosing spondylitis (AS) is a chronic inflammatory disease primarily affecting the spine and sacroiliac joint. Anoikis is a newly recognized form of programmed cell death that has been implicated in the progression of several diseases. Thus, the objective of this research is to investigate the factors associated with anoikis that contribute to the progression of ankylosing spondylitis. Methods: Two datasets for AS were obtained from the gene expression omnibus database. By analyzing the differentially expressed genesbetween the AS and control groups, as well as the genes associated with anoikis,the differentially expressed genes related toanoikis (AnDEGs) were identified. Toinvestigate the immune cell infiltration in AS, the CIBERSORT method was employed. Gene set enrichment analysis was used to assessthe function of the genes. Using machine learning algorithms, a set of bub-anoikis-related genes was identified and used to classify 68 AS samples into two distinctsubclusters. We then performed Gene Ontology, Kyoto Encyclopedia of Genes and Genomes pathway enrichment, Gene Set Variation Analysis,and immune infiltration analysis to evaluate the characteristicsof thesesubclusters. Finally, to validate the results of our bioinformatic analysis, immunohistochemistry, proteomic analysis, and blood routine examinationswere conducted. Results: We identified a total of 11 AnDEGs,and using machine learning algorithms, four hub genes, namelyID2, DNMT1, EZR, and ITGB3, were further identified. The majorityof these hub genes exhibitedlower expression levelsin AS samples,and their expression was found to be negatively correlated with immune cell infiltration. Two distinct subclusters exhibiting differential expression of genes primarilyassociated with the immune response process were constructed. Immune signalingpathways were enrichedin cluster2. Consistently, the level of immune infiltration was higher in cluster2. Immunohistochemistry verified the differentialexpression of the hubgenes, and blood routine examination confirmed differences in immune infiltration between the two subclusters. However, proteomic analysis detected only differentialexpression of EZR in AS patients with hip arthropathy. Conclusion: The dysregulation of EZR appears to bethe crucialfactor associated with neutrophil extracellular traps(NETs) in ankylosing spondylitis patients with hip arthropathy.