#3802 role of neutrophil extracellular traps in diabetic kidney disease: data from integration of bulk rna and single-nucleus rna sequencing analyses

Abstract Background and Aims Neutrophil extracellular traps (NETs) is an important immune response against infections that is implicated in various immune-mediated conditions. Growing evidence shows that inflammation has key pathogenic contributions in diabetic kidney disease (DKD) but the role of NETs remains unclear. Method We evaluated the differentially expressed genes (DEGs) of NETs in human DKD using the bulk RNA sequencing datasets of kidney biopsy from DKD patients (GSE30528 and GSE30529). The candidate genes were further selected based on the machine learning algorithms (LASSO and SVM-RFE). The single-nucleus RNA sequencing (GSE195460 and GSE131882) and DKD bulk RNA sequencing (GSE30122) were used to validate the results. Receiver Operating Characteristics (ROC) curve were constructed to assess the diagnostic performance of the identified genes for DKD. Candidate drugs were further screened from the DSigDB database. Results Three characteristic genes (ITGAM, ITGB2 and TLR7) were selected using machine learning approach, which were all upregulated in human DKD glomerular and tubulointerstitial portion compared to healthy controls (all P <0.05). Single-cell analysis further demonstrated that ITGAM, ITGB2 and TLR7 were mainly overexpressed in leucocytes. ITGAM, ITGB2 and TLR7 expression in tubulointerstitial compartment also showed good diagnostic performance for DKD (ROC AUC 0.983, 0.975 and 0.958 respectively). Candidate drugs that target ITGAM, ITGB2 or TLR7 genes in DKD include ropivacaine, lidocaine and sulfasalazine. Conclusion ITGAM, ITGB2 and TLR7 may serve as biomarkers for DKD and drugs that target these genes may have therapeutic potential for DKD.