Molecular diagnosis of polycystic ovary syndrome in obese and non-obese women by targeted plasma miRNA profiling

Abstract

Objective: Polycystic ovary syndrome (PCOS) is diagnosed based on the cli nical signs, but its presentation is heterogeneous and potentially confounded by concurrent conditions, such as obesity and insulin resistance. miRNA have recently emerged as putative pathophysiological and diagno stic factors in PCOS. However, no reliable miRNA-based method for molecular diagnosis of PCOS has been reported. The aim of this study was to develop a tool for accurate diagnosis of PCOS by targeted miRNA profiling of pl asma samples, defined on the basis of unbiased biomarker-finding analyses and biostatistical tools. Methods: A case-control PCOS cohort was cross-sectionally studied, incl uding 170 women classified into four groups: non-PCOS/lean, non-PCOS/obese, PCOS/lean, and PCOS/obese women. High-throughput miRNA analyses were performed in plasma, using NanoString technology and a 800 huma n miRNA panel, followed by targeted quantitative real-timePCR validation. Statistics were applied to define optim al normalization methods, identify deregulated biomarker miRNAs, and build classification algorithms, considering PCOS and obesity as major categories. Results: The geometric mean of circulating hsa-miR-103a-3p, hsa-miR-125 a-5p, and hsa-miR-1976, selected among 125 unchanged miRNAs, was defined as optimal reference for inter nal normalization (named mR3-method). Ten miRNAs were identified and validated after mR3-normalization as differentially expressed across the groups. Multinomial least absolute shrinkage and selection operator reg ression and decision-tree models were built to reliably discriminate PCOS vs non-PCOS, either in obese or non-obese wom en, using subsets of these miRNAs as performers. Conclusions: We define herein a robust method for molecular classification of PCOS based on unbiased identification of miRNA biomarkers and decision-tree protocols. This method al lows not only reliable diagnosis of non-obese women with PCOS but also discrimination between PCOS and obesity. Capsule: We define a novel protocol, based on plasma miRNA profiling, for molecular diagnosis of PCOS. This tool not only allows proper discrimination of the condition in non-obese women but also permits distinction between PCOS and obesity, which often display overlapping clinical presentat ions.