Re-programming of low-grade inflammatory monocytes

Purpose: The goals of this study are to compare the mircoRNA expression profiles from monocytes challegned with subclinical low dosage of LPS Methods: Primary murine bone marrow derived monocytes treated with or without 5 pg/ml LPS in complete culture medium. Harvested mircoRNAs were used to prepare libaray for subsequent sequencing, in triplicate, using HiSeq 1000. Sequences were assayed to ensure high quality for all samples using a custom R script. Reads were first trimmed for adaptor and quality using a combination of custom perl scripts and Btrim64. After trimming, trimmed fastq sequences were imported into mirAnalyzer for mapping, miRNA assembly and prediction, quantitation and differential expression. qRT–PCR validation was performed to validate selected microRNAs. Results: miRNA-seq data confirmed selective induction of a limited set of mircoRNAs in monocytes treated with subclinical super-low dose LPS. Conclusions: Our study revealed that there are selected miroRNAs preferentially induced in monocytes challenged with subclinical low dosage of LPS. microRNAs from monocytes cultured with or without 5 pg/ml LPS were harvested, prepared for sequencing, in triplicate, using HiSeq 1000.