Role of Disulfidptosis in the Local Inflammatory Response of Atopic Dermatitis

Understanding cell death pathways is critical for elucidating the mechanisms underlying inflammation in atopic dermatitis (AD). This study investigates the role of disulfidptosis, a novel form of programmed cell death, in AD pathogenesis.

RNA-seq datasets GSE193309 and GSE121212 from GEO were analyzed to examine 43 disulfidptosis-related genes. Differentially expressed genes (DEGs) were identified using the limma package. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to annotate biological functions and signaling pathways. Weighted Gene Co-expression Network Analysis (WGCNA) was applied to identify gene modules significantly associated with AD. A protein-protein interaction (PPI) network was constructed using STRING data, and further visualized and analyzed with Cytoscape to identify core mRNA-pathway relationships involving disulfidptosis. Key disulfidptosis-related mRNAs were validated in the dataset GSE121212. Following house dust mite (HDM) extract treatment, disulfidptosis was induced in HaCaT cells, with characteristic F-actin collapse visualized by confocal microscopy and redox imbalance confirmed by an increased NADP⁺/NADPH ratio. Expression of disulfidptosis-related genes was quantified via RT-qPCR.

A total of 4239, 3570, and 516 DEGs were identified between lesional skin (LS) and healthy control (HC), LS and non-lesional skin (NL), and NL and HC, respectively. WGCNA clustered these DEGs into 14 co-expression modules, five of which were significantly correlated with AD. Notably, the turquoise module showed the strongest association with LS and contained four disulfidptosis-related genes: ACTB, GYS1, SLC7A11, and MYH9. These four genes were consistently upregulated in LS compared to both NL and HC. Immunofluorescence staining showed F-actin contraction and membrane detachment in HDM-treated HaCaT cells, consistent with disulfidptotic morphology. A significant increase in the NADP⁺/NADPH ratio further supported disulfidptosis induction. qPCR confirmed upregulation of four key disulfidptosis-related genes (ACTB, GYS1, SLC7A11, and MYH9) in response to HDM exposure. Moreover, HDM treatment triggered a pronounced pro-inflammatory response, as evidenced by the elevated mRNA expression of cytokines IL-25, IL-33, TSLP, IL-6, and IL-8.

Our findings reveal an association between disulfidptosis-related gene signatures and AD pathogenesis, suggesting that this pathway may contribute to the inflammatory response observed in lesional skin.