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

• Published in: Scientific reports Vol. 14; no. 1; pp. 26436 - 14
• 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: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 02-11-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 631/337; 692/699/3161; Aniridia; Cell differentiation; Chromosome 5; Cornea; Corneal inflammation; Epithelial cells; Extracellular signal-regulated kinase; Eye diseases; Humanities and Social Sciences; Keratitis; Lipopolysaccharides; MiR-204-5p; multidisciplinary; PAX6; Pax6 protein; Science; Science (multidisciplinary); Stem cells; MiR-204-5p; PAX6; Keratitis; Corneal inflammation; Aniridia
• 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.