WTAP Promotes the Excessive Proliferation of Airway Smooth Muscle Cells in Asthma by Enhancing AXIN1 Levels Through the Recognition of YTHDF2.

IF 2.1 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemical Genetics Pub Date : 2024-10-25 DOI:10.1007/s10528-024-10947-7

Xueli Chen, Li Dai

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引用次数: 0

Abstract

Asthma is a common chronic respiratory disease in children, the incidence rate of which has increased in recent years. Wilms tumour 1-associated protein (WTAP) is an N6-methyladenosine (m6A) methyltransferase. The purpose of this study was to explore the specific mechanism of WTAP in asthma progression, and clarify the intricate interplay between m6A modifications, WTAP, AXIN1, and their collective impact on airway smooth muscle cells (ASMCs) proliferation in asthma. Platelet-derived growth factor-BB (PDGF-BB)-treated ASMCs were used to establish an asthma model in vitro. The cell phenotype was tested using CCK-8, transwell, and wound healing assays. The expression of the Wnt signalling pathway was detected by western blotting. In addition, the relationship between WTAP/YTDHF2 and AXIN1 was assessed by a double luciferase reporter assay. Actinomycin D treatment and RT‒qPCR assays were performed to determine the mRNA stability of AXIN1. We found that WTAP was significantly increased in PDGF-BB-treated ASMCs. Knockdown of WTAP inhibited the excessive cell viability and migration of ASMCs induced by PDGF-BB. Furthermore, WTAP knockdown increased AXIN1 levels and inhibited the Wnt signalling pathway. Furthermore, WTAP knockdown decreased the m6A levels and enhanced the mRNA stability of AXIN1. WTAP overexpression showed the opposite effect. In addition, YTHDF2 was demonstrated to be the reader that recognizes the WTAP-mediated m6A modification of AXIN1. YTHDF2 knockdown enhanced the mRNA stability of AXIN1 and reversed the effect of WTAP overexpression on PDGF-BB-treated ASMCs. WTAP knockdown inhibited the excessive cell viability and migration of ASMCs by enhancing the m6A levels of AXIN1, which was further recognized by YTHDF2. The upregulation of AXIN1 mediated by the WTAP/YTHDF2 axis further inhibited the Wnt signalling pathway. Our study provides a new method for the treatment of asthma. This work not only deepens our understanding of the molecular underpinnings of asthma but also identifies potential therapeutic targets for the development of novel treatments aimed at inhibiting ASMC proliferation and alleviating asthma symptoms.

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WTAP 通过识别 YTHDF2 提高 AXIN1 水平，从而促进哮喘患者气道平滑肌细胞的过度增殖。

哮喘是儿童常见的慢性呼吸道疾病，近年来发病率有所上升。Wilms tumor 1-associated protein（WTAP）是一种 N6-甲基腺苷（m6A）甲基转移酶。本研究旨在探索 WTAP 在哮喘进展中的具体机制，并阐明 m6A 修饰、WTAP、AXIN1 之间错综复杂的相互作用及其对哮喘中气道平滑肌细胞（ASMCs）增殖的共同影响。经血小板衍生生长因子-BB（PDGF-BB）处理的气道平滑肌细胞在体外建立了哮喘模型。细胞表型通过 CCK-8、跨孔和伤口愈合试验进行检测。Wnt 信号通路的表达通过 Western 印迹法进行检测。此外，还通过双荧光素酶报告实验评估了 WTAP/YTDHF2 和 AXIN1 之间的关系。通过放线菌素 D 处理和 RT-qPCR 检测来确定 AXIN1 的 mRNA 稳定性。我们发现，WTAP 在 PDGF-BB 处理的 ASMC 中明显增加。敲除 WTAP 可抑制 PDGF-BB 诱导的 ASMC 细胞过度存活和迁移。此外，敲除 WTAP 还能提高 AXIN1 的水平，抑制 Wnt 信号通路。此外，WTAP 的敲除降低了 m6A 的水平，增强了 AXIN1 的 mRNA 稳定性。WTAP 的过表达则显示出相反的效果。此外，YTHDF2被证明是识别WTAP介导的AXIN1 m6A修饰的阅读器。敲除 YTHDF2 可增强 AXIN1 的 mRNA 稳定性，并逆转 WTAP 过表达对 PDGF-BB 处理的 ASMC 的影响。WTAP敲除通过提高AXIN1的m6A水平抑制了ASMCs过高的细胞活力和迁移，而YTHDF2则进一步识别了AXIN1的m6A水平。WTAP/YTHDF2 轴介导的 AXIN1 上调进一步抑制了 Wnt 信号通路。我们的研究为治疗哮喘提供了一种新方法。这项工作不仅加深了我们对哮喘分子基础的理解，还为开发旨在抑制 ASMC 增殖和缓解哮喘症状的新型疗法确定了潜在的治疗靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemical Genetics 生物-生化与分子生物学

CiteScore

3.90

自引率

0.00%

发文量

133

审稿时长

4.8 months

期刊介绍： Biochemical Genetics welcomes original manuscripts that address and test clear scientific hypotheses, are directed to a broad scientific audience, and clearly contribute to the advancement of the field through the use of sound sampling or experimental design, reliable analytical methodologies and robust statistical analyses. Although studies focusing on particular regions and target organisms are welcome, it is not the journal’s goal to publish essentially descriptive studies that provide results with narrow applicability, or are based on very small samples or pseudoreplication. Rather, Biochemical Genetics welcomes review articles that go beyond summarizing previous publications and create added value through the systematic analysis and critique of the current state of knowledge or by conducting meta-analyses. Methodological articles are also within the scope of Biological Genetics, particularly when new laboratory techniques or computational approaches are fully described and thoroughly compared with the existing benchmark methods. Biochemical Genetics welcomes articles on the following topics: Genomics; Proteomics; Population genetics; Phylogenetics; Metagenomics; Microbial genetics; Genetics and evolution of wild and cultivated plants; Animal genetics and evolution; Human genetics and evolution; Genetic disorders; Genetic markers of diseases; Gene technology and therapy; Experimental and analytical methods; Statistical and computational methods.