Study of the microRNA expression profile dysregulation by hydrogen peroxide on the retinal pigment epithelium cells: role of miR-205-5p
Year: 2021
Type: dissertation
Abstract: Age related macular degeneration (ADM) and diabetic \nretinopathy (DR) are common retina-related diseases leading to \nblindness. The retinal pigment epithelium (RPE) is essential for the \nvision, in fact is well documented that oxidative stress (OS) generated \nin RPE and choroid neovascularization are related to the origin of these \ndisorders. The study of circulating miRNAs, small non-codding RNA, \nis opening new possibilities in terms of diagnosis and therapeutics. \nmiRNAs can travel inside small Extracellular Vesicles (sEVs) playing \nan important role in intracellular communication. Concretely, miR205-5p is involved in VEGF-related angiogenesis and seems to regulate \nassociated cell signaling pathways. \nOne of the aims of the present work is to establish a miRNA \nexpression profile of ARPE-19 cells and sEVs released from ARPE-19 \ncells under oxidative conditions (H2O2). Moreover, we want to evaluate \nthe role of the identified miRNAs and sEVs in the progression of \npathological processes related to retinal disorders. \nThe miRNA expression profile was carried out using SeraMir \nProfiler that includes 384 miRNAs whose expression was analyzed in \nARPE-19 cells and sEVs released by ARPE-19 cells, both in control \nconditions and treated with 600 M H2O2 for 24 hours. Additionally, \nthe role of miRNAs and sEVs in angiogenic processes was studied \nusing matrigel assays with HUVEC cells. \nWe identify a different miRNA expression profile between \noxidative stressed and control ARPE-19 cells (Cell miRNAs) and the \nsEVs (sEV miRNAs) released by them. As a result, 306 out of 384 Cell miRNAs were detected by the array and 7 were significantly overexpressed in H2O2-treated cells. Moreover, 218 miRNAs could be \ndetected in control and H2O2-induced sEVs and only 2 of them were \nsignificantly under-expressed in H2O2-induced sEVs. Among the \ndysregulated Cell miRNAs we found the miR-205-5p. Our results \nindicate that miR-205-5p is modulated by OS and regulates VEGFAangiogenesis. We also found the role of 600 M H2O2-sEVs on \noxidative stress induction, cell death increasing and angiogenesis \nstimulation in healthy ARPE-19 cells. \nThe miRNA dysregulation observed suggest their implication \nin retinal disorders after oxidative challenge. Among the dysregulated \nmiRNAs, miR-205-5p is proposed as a candidate against eye-related \nproliferative diseases. Therefore, results herein suggest the influence of \nsEV in neighboring cells and the possible utility of sEV miRNAs as \nbiomarkers.
Author: Oltra Sanchis, María
Language: English
Subfield: Ophthalmology
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W7062029347