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

• Published in: Scientific reports 2024-11, Vol.14 (1), p.26436-14, Article 26436
• Main Authors: Abbasi, Mojdeh, Amini, Maryam, Moustardas, Petros, Gutsmiedl, Quirin, Javidjam, Dina, Suiwal, Shweta, Seitz, Berthold, Fries, Fabian N., Dashti, Ava, Rautavaara, Yedizza, Stachon, Tanja, Szentmáry, Nóra, Lagali, Neil
• Format: Article
• Language: English
• Subjects: 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
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-76654-w

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.