MiRNA profiling in the aqueous humour in patients with Fuch's endothelial corneal dystrophy

Abstract

Aims: The aim of the study was to determine and compare the expression profiles of miRNA molecules in the aqueous humour (AH) of patients with Fuch's endothelial corneal dystrophy (FECD) and patients with normal corneal endothelium undergoing cataract surgery.Methods: The study consisted of 20 Caucasian patients (10 FECD and 10 control patients) who underwent comprehensive ophthalmology examination, thus routine corneal transplant or cataract surgery. 100 μL of AH were obtained from the anterior chamber at the beginning of the surgery with special care to avoid contamination with blood tears. The total RNA was isolated from the AH samples using an miRNeasy Serum/Plasma kit according to the manufacturer's instructions. The RNA concentration was determined by using NanoDrop2000c spectrophotometer. In addition, RNA analysis was performed using Agilent Bioanalyzer2100 and PicoRNA Kit according to the manufacturer's protocol. A microarray system (GeneChip miRNA 4.0 Array chip, Affymetrix) was used to determine the miRNA expression profiles. The RNA preparation and hybridization were performed according to the manufacturer's protocol, with one modification to extend the hybridization time to 42 h. The gene chips were scanned with an Affymetrix GeneChip Scanner 3000.Results: We found significant overexpression of hsa‐miR‐4532, hsa‐miR‐6800‐5p, hsa‐miR‐3621, hsa‐miR‐4270, hsa‐miR‐548ac, hsa‐miR‐328‐5p, hsa‐miR‐6771 ‐5p, hsa‐miR‐6858‐5p, hsa‐miR‐548a‐3p, hsa‐miR‐6069, hsa‐miR‐6716‐5p, hsa‐miR‐7151‐3p, hsa‐miR‐5189‐5p, hsa ‐miR‐4279, hsa‐miR‐1233‐5p, hsa‐miR‐455‐3p in patients with FECD compared to those patients with normal corneal endothelium. Reduced expression of hsa‐miR‐3613‐3p, hsa‐miR‐1298‐3p, hsa‐miR‐16‐2‐3p, hsa‐miR‐4668‐5p was observed in patients with FECD in addition to the control group.Conclusions: The analysis of miRNA expression profiles and their application as potential biomarkers will facilitate and accelerate the diagnosis of FECD, as well as enable the development of individualized molecularly targeted therapy.