Profiling of CD14+ monocytes from humans with Type diabetes and without diabetes [RNA-Seq]

Monocyte activation by high glucose and free fatty acids promotes inflammation implicated in vascular complications associated with Type 2 diabetes (T2D). Emerging evidence shows that long non coding RNA (lncRNA)s regulate inflammation, but their role in T2D induced monocyte dysfunction is unclear. To examine this, we profiled the transcriptome of CD14+ monocytes from volunteers with T2D and without diabetes (n=5 each) using strand-specific RNA-seq on Illumina HiSeq 2500. Our study identified several differentially regulated lncRNAs along with coding genes involved in monocyte functions related to inflammation and monocytosis. Following approved IRB protocols (at City of Hope and Baylor College of Medicine) informed consent was obtained from volunteers and fasting blood (15 ml) was collected from Control volunteers and volunteers with T2D at the Baylor College of Medicine. PBMC were isolated using Ficoll-Hypaque gradient (Sigma) and CD14+ monocytes were isolated by cell depletion with magnetic beads (Miltenyi Biotech, San Diego, CA). Total RNA was isolated from CD14+ monocytes using RNeasy mini kit and on-column DNase I digestion to eliminate genomic DNA (Qiagen). Total RNA from CD14+ human monocytes was subjected to strand-specific RNA-seq on Hi-Seq 2500 platform (Illumina, San Diego, CA).