Journal Article

Effects of miR-204-5p modulation on PAX6 regulation and corneal inflammation

Javidjam, Dina, Stachon, Tanja, Suiwal, Shweta, Gutsmiedl, Quirin, Amini, Maryam, Moustardas, Petros, Lagali, Neil, Abbasi, Mojdeh, Dashti, Ava, Szentmáry, Nóra, Rautavaara, Yedizza, Fries, Fabian N., Seitz, Berthold

2024

Overview

Congenital aniridia is a rare eye disease characterized by loss of PAX6 protein leading to aniridia-associated keratopathy that significantly reduces vision. The miR-204-5p is a possible regulator of PAX6 function and here we evaluate its effect in multiple in vitro and in vivo models. In vitro, miR-204-5p overexpression suppressed vascular factor ANGPT1 in human limbal stem cells (T-LSC) and Pax6-knockdown LSC (mut-LSC), and in primary human limbal epithelial cells (LEC) at the gene and protein levels and following LPS stimulation. However, miR-204-5p inhibited VEGFA expression only in mut-LSCs and LPS-stimulated LEC. Also, miR-204-5p increased PAX6 expression in mut-LSC and differentiated corneal epithelial cells, but not in LEC. Topical miR-204-5p after LPS-induced keratitis in mice failed to suppress Vegfa , Angpt1 , Il-1β , and Tnf-α or rescue Pax6 levels in contrast to in vitro results, although it significantly reduced the inflammatory infiltrate in the cornea. In Pax6 Sey/+ aniridia mice, miR-204-5p did not rescue PAX6 levels or suppress Vegfa , Angpt1 , or inhibit the ERK1/2 pathway. While short-term miR-204-5p treatment effectively suppresses VEGFA and ANGPT1 and enhances PAX6 expression in multiple corneal epithelia, effects are variable across primary and immortalized cells. Effects of longer-term in vivo treatment, however, require further study.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 631/337 / 692/699/3161 / Animals / Aniridia / Aniridia - genetics / Aniridia - metabolism / Aniridia - pathology / Cell differentiation / Chromosome 5 / Cornea / Corneal inflammation / Epithelial cells / Extracellular signal-regulated kinase / Eye diseases / Gene Expression Regulation / Humanities and Social Sciences / Humans / Inflammation - genetics / Inflammation - metabolism / Inflammation - pathology / Keratitis / Lipopolysaccharides / Mice / MicroRNAs - genetics / MicroRNAs - metabolism / MiR-204-5p / multidisciplinary / PAX6 / Pax6 protein / PAX6 Transcription Factor - genetics / PAX6 Transcription Factor - metabolism / Science / Science (multidisciplinary) / Stem cells / Vascular Endothelial Growth Factor A - genetics / Vascular Endothelial Growth Factor A - metabolism