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METTL14-mediated depression of NEIL1 aggravates oxidative damage and mitochondrial dysfunction of lens epithelial
Lihua Kang1, Sijie Bao1, Pengfei Li1
1Eye Institute, Affiliated Hospital of Nantong University, Medical School of Nantong University, Nantong 226001, China.
Abstract:
Abnormal base excision repair (BER) pathway and N6-methyladenosine (m6A) of RNA have been proved to be significantly related to age-related cataract (ARC) pathogenesis. However, the relationship between the Nei Endonuclease VIII-Like1 (NEIL1) gene (a representative DNA glycosylase of BER pathway) and its m6A modification remains unclear. Here, we showed that the expression of NEIL1 was decreased in the ARC anterior lens capsules and H2O2-stimulated SRA01/04 cells. Our findings demonstrated that ectopic expression of NEIL1 alleviated DNA oxidative damage, apoptosis and mitochondrial dysfunction through disturbing KEAP1/NRF2 interaction. Furthermore, silencing NEIL1 aggravated H2O2-induced lens opacity, whereas ML334 could mitigate lens cloudy ex vitro in rat lenses. Besides, intravitreal injection of AAV2-NEIL1 alleviated lens opacity in Emory mice in vivo. Mechanistically, the N(6)-Methyladenosine (m6A) methyltransferase-like 14 (METTL14) was identified as a factor in promoting m6A modification of NEIL1, which resulted in the recruitment of YTHDF2 to recognize and impair NEIL1 RNA stability. Collectively, these findings highlight the critical role of the m6A modification in NEIL1 on regulating oxidative stress and mitochondrial homeostasis through KEAP1/NRF2 pathways, providing a new way to explore the pathogenesis of ARC.
Insights
The Nei Endonuclease VIII-Like1 (NEIL1) gene
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- Age-related cataract (ARC) pathogenesis involves abnormal base excision repair (BER) and RNA N6-methyladenosine (m6A) modification.
- The specific role of the NEIL1 gene, a key BER component, and its m6A modification in ARC remains largely unknown.
Purpose of the Study:
- To investigate the function of NEIL1 and its m6A modification in the pathogenesis of age-related cataract.
- To elucidate the underlying molecular mechanisms involving oxidative stress and mitochondrial homeostasis.
Main Methods:
- Quantitative analysis of NEIL1 expression in ARC tissues and cells.
- Investigating the effect of NEIL1 modulation on oxidative damage, apoptosis, and mitochondrial function.
- Utilizing in vitro and in vivo models (rat lenses, Emory mice) to assess NEIL1's therapeutic potential.
- Identifying m6A modification regulators (METTL14, YTHDF2) and their impact on NEIL1 RNA stability.
Main Results:
- NEIL1 expression was decreased in ARC anterior lens capsules and H2O2-treated cells.
- Ectopic NEIL1 expression reduced oxidative damage, apoptosis, and mitochondrial dysfunction by modulating the KEAP1/NRF2 pathway.
- NEIL1 silencing exacerbated H2O2-induced lens opacity, while ML334 and AAV2-NEIL1 treatments mitigated opacity in vivo and ex vivo.
- METTL14 promotes m6A modification of NEIL1, leading to YTHDF2-mediated degradation and reduced RNA stability.
Conclusions:
- m6A modification of NEIL1 plays a critical role in regulating oxidative stress and mitochondrial homeostasis via the KEAP1/NRF2 pathway in ARC.
- NEIL1 and its m6A modification represent a potential therapeutic target for age-related cataract.
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