Clinical & Experimental Ophthalmology

Biography

Biography: Maria Dolores Pinazo Durán

Abstract

Purpose: To utilize the tear film as biological source of micro RNAs (miRNAs) in order to analyze its profile in relation to primary open angle glaucoma (POAG). Material and methods: A total of 30 participants (POAG patients (GG; n=15) and matched healthy controls (CG; n=15) were interviewed, ophthalmologically examined and extracted their reflex tears atraumatically from the inferior meniscus (20-30 microL) with a Pasteur micropipette that were stored at -80º until processing. The mercury TM RNA isolation kit (Exiqon, Denmark) was used for RNA extraction. The SPSS 15.0 program was used for statistical analysis by integrating all variables. Results: Mean age of the participants was 70 + 2 years in the GG vs 68 + 2 years in the CG. We extracted RNA from the pooled tear film samples and its quality was evaluated using Agilent Bioanalyzer 2100. Then, pure RNA (< 200 nucleotides) was utilized to generate a sequencing library for small RNAs. It was obtained significantly higher purified RNA in tears from the GG as compared to the CG [11 + 1 ng/mL vs 8 + 2 ng/mL of pure RNA respectively (p<0.001)]. By next generation sequencing it has been detected a high variety of matured miRNAs with a differential expression profile in tears from both groups of participants. Conclusions: Tear film sampling is a non-invasive, relatively simple and efficient process for glaucoma research. The miRNAs obtained from tears can be used for quantitative and qualitative assays to improve knowledge on the molecular basis of POAG. The possibility remain that specific miRNAs can be used as biomarkers for better monitoring the diagnosis and therapy of the glaucomatous patients.