TY - JOUR T1 - Small RNA-sequence analysis of plasma-derived extracellular vesicle miRNAs in smokers and patients with chronic obstructive pulmonary disease as circulating biomarkers. AU - Sundar,Isaac Kirubakaran, AU - Li,Dongmei, AU - Rahman,Irfan, Y1 - 2019/11/07/ PY - 2019/01/14/received PY - 2019/09/24/revised PY - 2019/10/20/accepted PY - 2019/11/26/entrez PY - 2019/11/26/pubmed PY - 2019/11/26/medline KW - COPD KW - Extracellular vesicles KW - biomarker KW - miRNAs KW - next-generation sequencing SP - 1684816 EP - 1684816 JF - Journal of extracellular vesicles JO - J Extracell Vesicles VL - 8 IS - 1 N2 - Extracellular vesicles (EVs) play a vital role in normal lung physiology to maintain homeostasis in the airways via intercellular communication. EVs include exosomes and microvesicles, and are characterized by their phospholipid bilayers. EVs have been recognized as novel circulating biomarkers of disease, which are released by different cell types. In this study, we used different EV isolation and purification methods to characterize the plasma-derived EV miRNAs from non-smokers, smokers and patients with COPD. A small RNA sequencing (RNA-seq) approach was adapted to identify novel circulating EV miRNAs. We found that plasma-derived EVs from non-smokers, smokers and patients with COPD vary in their size, concentration, distribution and phenotypic characteristics as confirmed by nanoparticle tracking analysis, transmission electron microscopy, and immunoblot analysis of EV surface markers. RNA-seq analysis confirmed the most abundant types of small RNAs, such as miRNAs, tRNAs, piRNAs snRNAs, snoRNAs and other biotypes in plasma-derived EVs. We mainly focused on miRNAs as novel biomarkers in smokers and patients with COPD for further analysis. Differential expression by DESeq2 identified distinct miRNA profiles (up-regulated: miR-22-3p, miR-99a-5p, miR-151a-5p, miR-320b, miR-320d; and down-regulated: miR-335-5p, miR-628-3p, miR-887-5p and miR-937-3p) in COPD versus smokers or non-smokers in a pairwise comparison. Gene set enrichment analysis (GSEA) of differentially expressed miRNAs revealed the top pathways, gene ontology and diseases associated with smokers and patients with COPD. We selectively validated miRNAs in EVs isolated from BEAS-2B cells treated with cigarette smoke extract by quantitative PCR analysis. For the first time, we report that plasma-derived EV miRNAs are novel circulating pulmonary disease biomarkers. Thus, molecular profiling of EV miRNAs has great translational potential for the development of biomarkers that may be used in the diagnosis, prognosis, and therapeutics of COPD. SN - 2001-3078 UR - https://www.unboundmedicine.com/medline/citation/31762962/Small_RNA_sequence_analysis_of_plasma_derived_extracellular_vesicle_miRNAs_in_smokers_and_patients_with_chronic_obstructive_pulmonary_disease_as_circulating_biomarkers_ DB - PRIME DP - Unbound Medicine ER -