YTHDF1-Mediated m6A Modification of NREP Promotes Corneal Fibrosis via TGF-β-Smad Signaling

Area of Science:

• Ophthalmology
• Molecular Biology
• Cell Biology

Background:

• Corneal fibrosis is a leading cause of vision loss with limited treatment options.
• Neuronal regeneration-related protein (NREP) and its regulator YTHDF1 are implicated in cellular processes.
• N6-methyladenosine (m6A) modification plays a role in gene regulation.

Purpose of the Study:

• To investigate the role of NREP, regulated by YTHDF1 and m6A, in promoting corneal fibrosis.
• To explore the underlying molecular mechanisms of NREP in keratocyte function.

Main Methods:

• Established a mouse alkali burn model and in vitro keratocyte culture system.
• Assessed corneal fibrosis, NREP, and m6A levels using RT-qPCR, western blotting, and other assays.
• Utilized siRNA to downregulate NREP and YTHDF1, and a specific YTHDF1 inhibitor (SKLB-Y13).

Main Results:

• Corneal fibrosis peaked at day 14 post-injury with elevated NREP levels.
• NREP knockdown reduced keratocyte proliferation, migration, and TGF-β1-induced α-SMA expression.
• YTHDF1 depletion mimicked these effects, reduced NREP protein, and impaired TGF-β-Smad signaling.

Conclusions:

• YTHDF1 promotes corneal fibrosis via m6A-dependent NREP translation.
• This process involves potentiation of TGF-β-Smad signaling and myofibroblast transdifferentiation.
• YTHDF1 represents a potential therapeutic target for fibrotic corneal diseases.