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METTL3-deficiency Suppresses Neural Apoptosis to Induce Protective Effects in Cerebral I/R Injury via Inhibiting RNA m6A Modifications: A Pre-clinical and Pilot Study

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Abstract

N6-Methyladenosine (m6A) RNA methylation involves in regulating the initiation, progression and aggravation of cerebral ischemia-reperfusion (I/R) injury, however, the detailed functions and mechanisms by which m6A drives cerebral I/R injury are not fully understood. This study found that methyltransferase-like 3 (METTL3) m6A-dependently regulated cerebral I/R injury trough regulating a novel LncRNA ABHD11-AS1/miR-1301-3p/HIF1AN/HIF-1α axis. Specifically, the middle cerebral artery occlusion (MCAO)/reperfusion mice models and glucose deprivation (OGD)/reoxygenation (RX) astrocyte cell models were respectively established, and we verified that METTL3, ABHD11-AS1 and HIF1AN were upregulated, whereas miR-1301-3p and HIF-1α were downregulated in both MCAO/reperfusion mice tissues and OGD/RX astrocytes. Mechanical experiments confirmed that METTL3 m6A dependently increased stability and expression levels of ABHD11-AS1, and elevated ABHD11-AS1 sponged miR-1301-3p to upregulate HIF1AN, resulting in the downregulation of HIF-1α. Moreover, silencing of METTL3 rescued MCAO/reperfusion and OGD/RX-induced oxidative stress-associated cell apoptosis and cell cycle arrest in both mice brain tissues in vivo and the mouse primary astrocytes in vitro, which were abrogated by overexpressing ABHD11-AS1 and downregulating miR-1301-3p. Taken together, our study firstly reported a novel METTL3/m6A/ ABHD11-AS1/miR-1301-3p/HIF1AN/HIF-1α signaling cascade in regulating the progression of cerebral I/R injury, and future work will focus on investigating whether the above genes can be used as biomarkers for the treatment of cerebral I/R injury by performing clinical studies.

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  1. Department of Neurology, The Eighth Affiliated Hospital, Sun Yat-Sen University, Shennanzhong Road 3025, Shenzhen, 518033, Guangdong, China

    Gang Huang, Yuda Qiu, Yafei Fan & Jianfeng Liu

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Contributions

Gang Huang designed the experiments, conducted the investigations and drafted the manuscript. Yuda Qiu and Yafei Fan helped to collect, analyze and visualize the data, and they also provided technical support for this study. Jianfeng Liu was the correspondence author who provided guidance for this study, and designed the experiments, drafted the manuscript and proofread the final version of the paper for potential publication.

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11064_2023_4015_MOESM1_ESM.jpg

Supplementary Figure S1. The mRNA levels of HIF1AN, HIF-1α, Nox4 and miR-1201-3p were respectively examined by performing Real-Time qPCR analysis in the (A-D) mice astrocytes and (E-H) mice brain tissues. Each experiment repeated for at least 3 times, and *P < 0.05 was considered as statistical significance.

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Huang, G., Qiu, Y., Fan, Y. et al. METTL3-deficiency Suppresses Neural Apoptosis to Induce Protective Effects in Cerebral I/R Injury via Inhibiting RNA m6A Modifications: A Pre-clinical and Pilot Study. Neurochem Res 49, 85–98 (2024). https://doi.org/10.1007/s11064-023-04015-6

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