Transcriptomics of Type 2 Diabetic and Healthy Human Neutrophils

Abstract

ABSTRACTObjectivesChronic inflammatory diseases, including diabetes and cardiovascular disease, are heterogeneous and often co-morbid, with increasing global prevalence. Uncontrolled type 2 diabetes (T2D) can result in severe inflammatory complications. As neutrophils are essential to inflammation, we conducted RNA-seq transcriptomic analyses to investigate the association between neutrophil gene expression and T2D phenotype. Further, as specialized pro-resolving lipid mediators, including resolvin E1 (RvE1), can actively resolve inflammation, we further surveyed the impact of RvE1 on isolated neutrophils.MethodsCell isolation and RNA-seq analysis of neutrophils from N=11 T2D and N=7 healthy individuals with available clinical data was conducted. Additionally, cultured neutrophils (N=3 T2D, N=3 healthy) were perturbed with increasing RvE1 doses (0nM, 1nM, 10nM, or 100nM) prior to RNA-seq. Data was evaluated through a bioinformatics pipeline including pathway analysis andpost hocfalse-discovery rate (FDR)-correction.ResultsWe observed significant differential expression of 50 genes between T2D and healthy neutrophils (p<0.05), including decreased T2D gene expression in inflammatory- and lipid-related genesSLC9A4, NECTIN2andPLPP3(p<0.003). RvE1 treatment induced dose-dependent differential gene expression (uncorrected p<0.05) across groups, including 59 healthy and 216 T2D neutrophil genes. Comparing T2D to healthy neutrophils, 1097 genes were differentially expressed across RvE1 doses, including two significant genes,LILRB5andAKR1C1, involved in inflammation (p<0.05).ConclusionsInflammatory- and lipid-related genes were differentially expressed between T2D and healthy neutrophils, and RvE1 dose-dependently modified gene expression in both groups. Unraveling the mechanisms regulating abnormalities in diabetic neutrophil responses could lead to better diagnostics and therapeutics targeting inflammation and inflammation resolution.