Transcriptomics of Type 2 Diabetic and Healthy Human Neutrophils

Abstract

Objectives: Chronic 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. Methods: Cell 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 and post hoc false-discovery rate (FDR)-correction. Results: We 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 genes SLC9A4, NECTIN2 and PLPP3 (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, LILRB5 and AKR1C1, involved in inflammation (p<0.05). Conclusions: Inflammatory- 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.

Inflammatory Networks Linking Oral Microbiome with Systemic Health and Disease

“…Neutrophils, normally present in periodontal tissues with sentinel capacity, are key immune cells for initial inflammation and its resolution, and neutrophil abnormalities—including impaired adhesion, cytokine signaling, and phagocytosis—are becoming increasingly recognized as being important to chronic inflammatory disease development (Alba-Loureiro et al 2007; Curtis et al 2011; Hotamisligil 2017; Serhan and Levy 2018; Feehan, Dalli, and Gilroy 2019). A recent transcriptomics study showed that chronic inflammation in type 2 diabetes correlated with reduced neutrophil gene expression as compared with healthy patients (Kleinstein et al 2019), similar to observations in gut dysbiosis (Hunt et al 2008). The main role of neutrophils in the inflammatory response is to clear tissue debris or microbes, which is partially completed by releasing neutrophil extracellular traps (NETs; Silvestre-Roig et al 2019).…”

“…This interaction between bacteria and the host immune response is critical: it is now understood that host variation in inflammatory genes, including IL-17, can increase the risk of periodontitis and systemic diseases (Bedoya et al 2013; Dutzan et al 2018; Borilova Linhartova et al 2019). Furthermore, we have shown evidence for dysregulation of neutrophil gene expression and cytokines in type 2 diabetes as compared with health (Kleinstein et al 2019). Dissecting interaction between specific signals from bacteria and inflammation is key to understanding the development of chronic inflammatory diseases.…”

Exploring the role of non-coding RNAs as potential candidate biomarkers in the cross-talk between diabetes mellitus and Alzheimer’s disease

“…It is found in the brain and neuronal cells and is necessary for proper cell junction formation. The expression of this protein was found to be lower in T2DM patients ( Kleinstein et al, 2019 ).…”