alkbh5介导的m6A去甲基化通过增强PELI2 mRNA的稳定性来促进皮肤伤口的再上皮化。

IF 5 3区 医学 Q2 IMMUNOLOGY
Xin Huang, Yixuan Zhao, Daiming Liu, Shuchen Gu, Yunhan Liu, Yimin Khoong, Shenying Luo, Zewei Zhang, Wenzheng Xia, Meng Wang, Hsin Liang, Minxiong Li, Qingfeng Li, Tao Zan
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引用次数: 0

摘要

背景:伤口再上皮化受损导致皮肤屏障重建功能障碍。近年来,n6 -甲基腺苷(m6A) RNA修饰已被证明参与RNA命运的决定,其畸变触发许多疾病的发病机制。然而,m6A在伤口再上皮化中的作用仍然是一个谜。方法:构建Alkbh5 - / -小鼠,研究Alkbh5消融后创面再上皮化率。采用甲基化RNA免疫沉淀测序(MeRIP-seq)和RNA-seq相结合的综合高通量分析方法鉴定ALKBH5的下游靶点。通过体外和体内救援实验验证下游靶点对alkbh5缺陷细胞或动物功能表型的作用。此外,通过RIP-qPCR、RNA下拉和RNA稳定性分析确定了相互作用的读取器蛋白和调控机制。结果:ALKBH5在创面边缘表皮特异性上调。在ALKBH5 - / -小鼠中,消融ALKBH5抑制角化细胞迁移并导致伤口再上皮化延迟。综合高通量分析发现,E3泛素蛋白连接酶PELI2是ALKBH5的下游靶点。与此同时,在体外和体内,外源性PELI2补充部分地挽救了角化细胞的迁移,加速了alkbh5缺陷细胞的再上皮化。就其机制而言,ALKBH5通过去除PELI2 mRNA上的m6A修饰,增强PELI2的稳定性,从而促进PELI2的表达,其机制依赖于ythdf2。结论:本研究确定了ALKBH5是伤口再上皮化的内源性促进剂,从而有利于开发一种重编程m6A靶向治疗难治性伤口的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

ALKBH5-mediated m<sup>6</sup>A demethylation fuels cutaneous wound re-epithelialization by enhancing PELI2 mRNA stability.

ALKBH5-mediated m<sup>6</sup>A demethylation fuels cutaneous wound re-epithelialization by enhancing PELI2 mRNA stability.

ALKBH5-mediated m<sup>6</sup>A demethylation fuels cutaneous wound re-epithelialization by enhancing PELI2 mRNA stability.

ALKBH5-mediated m6A demethylation fuels cutaneous wound re-epithelialization by enhancing PELI2 mRNA stability.

Background: Impaired wound re-epithelialization contributes to cutaneous barrier reconstruction dysfunction. Recently, N6-methyladenosine (m6A) RNA modification has been shown to participate in the determination of RNA fate, and its aberration triggers the pathogenesis of numerous diseases. Howbeit, the function of m6A in wound re-epithelialization remains enigmatic.

Methods: Alkbh5‒/‒ mouse was constructed to study the rate of wound re-epithelialization after ALKBH5 ablation. Integrated high-throughput analysis combining methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA-seq was used to identify the downstream target of ALKBH5. In vitro and in vivo rescue experiments were conducted to verify the role of the downstream target on the functional phenotype of ALKBH5-deficient cells or animals. Furthermore, the interacting reader protein and regulatory mechanisms were determined through RIP-qPCR, RNA pull-down, and RNA stability assays.

Results: ALKBH5 was specifically upregulated in the wound edge epidermis. Ablation of ALKBH5 suppressed keratinocyte migration and resulted in delayed wound re-epithelialization in Alkbh5‒/‒ mouse. Integrated high-throughput analysis revealed that PELI2, an E3 ubiquitin protein ligase, serves as the downstream target of ALKBH5. Concordantly, exogenous PELI2 supplementation partially rescued keratinocyte migration and accelerated re-epithelialization in ALKBH5-deficient cells, both in vitro and in vivo. In terms of its mechanism, ALKBH5 promoted PELI2 expression by removing the m6A modification from PELI2 mRNA and enhancing its stability in a YTHDF2-dependent manner.

Conclusions: This study identifies ALKBH5 as an endogenous accelerator of wound re-epithelialization, thereby benefiting the development of a reprogrammed m6A targeted therapy for refractory wounds.

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来源期刊
CiteScore
11.10
自引率
1.20%
发文量
45
审稿时长
11 weeks
期刊介绍: Inflammation and Regeneration is the official journal of the Japanese Society of Inflammation and Regeneration (JSIR). This journal provides an open access forum which covers a wide range of scientific topics in the basic and clinical researches on inflammation and regenerative medicine. It also covers investigations of infectious diseases, including COVID-19 and other emerging infectious diseases, which involve the inflammatory responses. Inflammation and Regeneration publishes papers in the following categories: research article, note, rapid communication, case report, review and clinical drug evaluation.
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