Identification and validation of PANoptosis-related genes in ankylosing spondylitis

Ankylosing spondylitis (AS) is a common immune inflammatory disease. PANoptosis, as a novel programmed cell death pathway, its mechanism of action in ankylosing spondylitis remains unclear. Therefore, this study aims to clarify the role of a novel programmed cell death pattern - PANoptosis - in the pathogenesis of ankylosing spondylitis (AS), and to screen and verify key genes (APRGS), providing new ideas for the diagnosis and treatment of AS. Based on multiple gene expression datasets (GSE25101, GSE73754, GSE11886, GSE134290) of AS patients and known gene libraries related to panoptosis, by integrating differential expression genes analysis (DEGs), weighted gene co-expression network analysis (WGCNA), protein-protein interaction network (PPI), and three machine learning algorithms (RF, LASSO-logistic regression, SVM), six core APRGs were identified: AIM2, TNF, IFNG, CASP8, ADAR, ALKBH5. These genes are significantly enriched in signaling pathways such as NOD-like receptors, TNF and p53, and exhibit excellent diagnostic efficacy for as (ROC analysis). Immune infiltration analysis revealed that as patients had characteristics such as an increase in activated NK cells and CD4+ t cells. In vivo validation was carried out by establishing a rat model of as induced by complete freesier adjuvant (CFA). HE staining showed that obvious inflammatory infiltration and abnormal ossification occurred in the sacroiliac joint of the model rats, and the levels of serum pro-inflammatory factors (TNF-α, IL-6) increased, and anti-inflammatory factors (IL-10, IL-4) significantly decreased. Molecular testing confirmed that the expression of RIPK1 protein was upregulated. The mRNA and protein expressions of AIM2, TNF, IFNG, CASP8 and ALKBH5 in core APRGs were significantly increased, while the expression of ADAR was decreased. Our research has clarified that PANoptosis drives chronic inflammation and bone destruction in as through the synergistic action of six key genes: AIM2, TNF, IFNG, CASP8, ADAR, and ALKBH5, involving NOD-like receptors, TNF, and p53 pathways. These genes are potential diagnostic markers and therapeutic targets for as, providing an important basis for the development of targeted intervention strategies.