D1.497 - An integrated multiomic insight to nonsteroidal anti-inflammatory drug-induced acute urticaria/angioedema

Cross-hypersensitivity to nonsteroidal anti-inflammatory drugs (NSAIDs) is the most frequent drug hypersensitivity type, with patients developing at least two reactions to NSAIDs belonging to different chemical groups. NSAID-induced urticaria/angioedema (NIUA) is the most common clinical entity. These reactions are thought to be bred by pharmacological cyclooxygenase-1 inhibition, leading to an impairment in eicosanoid biosynthesis; however, their underlying mechanisms are not fully understood. We aimed to investigate molecular processes involved in NIUA development through a multiomic integration analysis, which includes data from transcriptomics, proteomics, and metabolomics.

PBMCs were obtained from NIUA patients during a positive drug provocation test with acetylsalicylic acid (ASA; acute phase; n=5) and from NSAID-tolerant individuals after a full ASA dose intake (n=9). Transcriptomics was performed by RNAseq, proteomics by HPLC-MS in cell lysates and serum metabolomics by NMR. Coding RNA and protein abundances were normalised with DESeq2 and limma-voom, respectively. Data from metabolomics were processed with TopSpin and analysed by Chenomx Profiler. A multiomic integration model was generated with the DIABLO (Data Integration Analysis for Biomarker Discovery using Latent Variable Approaches for Omics Studies) algorithm, included in mixOmics. Model’s quality was measured using a permutation test, classification-error (CER) and ROC curves. Functional enrichment was done using clusterProfiler.

The final model (permutation p-value=0.009; ROC=1; CER=0.07) presented one component that included 45 coding-RNAs, 6 proteins, and 3 metabolites, showing high correlation between them (RNA-proteins, r=0.98; metabolites-proteins, r=0.83; RNA-metabolites, r=0.8). Most RNAs presented higher expression in NIUA patients, meanwhile most proteins displayed lower expression. Functional enrichment showed these molecules to be involved in immune mechanisms such as mast cell degranulation, lymphocyte and neutrophil migration/activation/chemotaxis, and FcεRI- and TLR-mediated stimulatory signalling pathways, with an important presence of proinflammatory transcription factors (JUN, DUSP1, MAFF) and cytokines (CXCL8, CD69, RSG1). Principal metabolites in the model were urea (upregulated in NIUA) and lactate (downregulated).

During the acute phase, NIUA patients showed an enhancement in some immunological processes, including proinflammatory pathways involving PBMCs, granulocytes activation and migration, and TLRs/FcεRI signalling. These results shed new light on the molecular basis of NIUA, delimiting a relevant immune context that may help to further elucidate NSAID-cross-hypersensitivity underlying mechanisms.