ALKBH5 Regulates Corneal Neovascularization by Mediating FOXM1 M6A Demethylation

Area of Science:

• Ophthalmology
• Molecular Biology
• Epigenetics

Background:

• Corneal neovascularization (CNV) is a significant cause of vision impairment.
• N6-methyladenosine (m6A) modification plays a role in various biological processes, including angiogenesis.
• The specific role of ALKBH5-mediated m6A demethylation in CNV remains underexplored.

Purpose of the Study:

• To investigate the regulatory function of ALKBH5 in CNV.
• To elucidate the underlying molecular mechanisms of ALKBH5 in CNV pathogenesis.
• To assess the therapeutic potential of targeting ALKBH5 for CNV treatment.

Main Methods:

• Established a mouse model of CNV using corneal alkali burns.
• Quantified m6A levels and mRNA expression of m6A-related enzymes.
• Utilized small interfering RNA to knockdown ALKBH5 in vivo and in vitro.
• Assessed cell proliferation, migration, and tube formation in human umbilical vein endothelial cells (HUVECs).
• Examined the interaction between ALKBH5 and FOXM1 using dual-luciferase and methylated RNA immunoprecipitation-qPCR assays.

Main Results:

• ALKBH5 expression was increased, while total m6A levels were reduced in CNV corneas and IL-6-induced HUVECs.
• ALKBH5 knockdown alleviated CNV, reduced inflammation, and inhibited HUVEC proliferation, migration, and tube formation.
• ALKBH5 depletion increased m6A levels on FOXM1 mRNA, leading to decreased FOXM1 expression.
• FOXM1 overexpression reversed the effects of ALKBH5 depletion.

Conclusions:

• ALKBH5-mediated m6A demethylation of FOXM1 is a key regulator of CNV progression.
• ALKBH5 acts as a pro-angiogenic factor in CNV.
• Targeting ALKBH5 presents a promising therapeutic strategy for managing corneal neovascularization.