Integrative Multi-Omics Analysis Provides Insights into the Molecular Mechanisms and Therapeutic Targets of Cicada Pupae Allergy

• Neuroactive ligand-receptor interaction and JAK-STAT signaling pathways have been identified to be associated with pruritus symptoms in cicada pupae allergy (CPA). • NOD-like receptor signaling pathway was the key mediator of CPA responses. • TYK2, sphingosine-1P and dihydrosphingosine-1P have been identified as key biomarkers for early CPA diagnosis. • Sphingolipid metabolism is the most significantly altered pathway between the CPA and HC groups. • The significant regulation of dihydrosphingosine-1P and COL4A2 gene is involved in the response to CPA. As edible insects become increasingly important in addressing growing global protein demand, cicada pupae are gaining attention for their high nutritional value and potential health benefits. However, the widespread consumption of cicada pupae has increased the number of allergies. This study aimed to first identify potential novel biomarkers of CPA and elucidate its pathogenesis mechanism. The immunologic profiles of 12 subjects we recruited were characterized and divided into CPA and healthy control (HC) groups based on clinical diagnosis results. By multi-omics analysis, the key genes, specific biomarkers and pathways associated with CPA symptoms were identified. The expression profiling of key genes and the levels of key proteins were validated using RT-qPCR and Elisa, respectively. A correlation analysis was performed to elucidate the pathogenesis of CPA. The results showed that the lymphocyte percentage and eosinophil percentage in CPA group were significantly higher than those in the HC group. Neuroactive ligand-receptor interaction and JAK-STAT signaling pathway were observed to contribute to pruritus and 9 genes associated with different CPA symptoms were identified. NOD-like receptor signaling pathway was the key mediator of CPA responses. Sphingolipid metabolism was the most significant perturbed pathway between CPA and HC groups. Notably, TYK2, sphingosine-1P, and dihydrosphingosine-1P were identified as potential biomarkers for CPA diagnosis. Multi-omics analysis revealed that the significant regulation of dihydrosphingosine-1P and COL4A2 gene is involved in the CPA response. These findings identified reliable diagnostic biomarkers and revealed potential therapeutic targets, providing novel insights into the diagnosis and treatment of CPA.