A Microarray, Validation, and Gene-Enrichment Approach for Assessing Differentially Expressed Circulating miRNAs in...

Obesity is a risk factor associated with cardiovascular diseases that may lead to heart failure (HF). However, in HF, overweight and obese patients have longer survival than underweight patients, a phenomenon known as the obesity paradox. MiRNAs play a fundamental role in gene regulation involved in obesity and HF. The main objective of this study was to identify and validate differentially expressed circulating miRNAs in HF-obese and HF-lean patients. This case-control study was carried out in two phases: discovery and validation. In the discovery phase, plasma samples from 20 HF patients and from 10 healthy controls were analyzed using the miRNA 4.0 Affymetrix GeneChip array. Differentially expressed miRNAs were ranked and selected for validation. In this phase, plasma miRNAs -451a, -22-3p, and -548ac from 80 patients and controls were analyzed by qPCR. Target analysis and functional enrichment analysis were performed. When comparing HF-lean and HF-obese groups compared to controls, miRNAs -451a and -22-3p were up-regulated in both discovery and validation phases, while -548ac was down-regulated in the discovery phase and up-regulated in the validation phase, indicating that miRNA changes are independent of obesity. These miRNAs regulate genes and different biological processes associated with metabolic, morphological, and functional outcomes.