ALKBH5-mediated m6A demethylation ameliorates extracellular matrix deposition in cutaneous pathological fibrosis

IF 7.9 1区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL Clinical and Translational Medicine Pub Date : 2024-09-04 DOI:10.1002/ctm2.70016

Ruoqing Xu, En Yang, Hsin Liang, Shenying Luo, Yunhan Liu, Yimin Khoong, Haizhou Li, Xin Huang, Yixuan Zhao, Tao Zan

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Abstract

Background

Elevated extracellular matrix (ECM) accumulation is a major contributing factor to the pathogenesis of fibrotic diseases. Recent studies have indicated that N6-methyladenosine (m⁶A) RNA modification plays a pivotal role in modulating RNA stability and contribute to the initiation of various pathological conditions. Howbeit, the precise mechanism by which m⁶A influences ECM deposition remains unclear.

Methods

In this study, we used hypertrophic scars (HTSs) as a paradigm to investigate ECM-related diseases. We focused on the role of ALKBH5-mediated m⁶A demethylation within the pathological progression of HTSs and examined its correlation with clinical stages. The effects of ALKBH5 ablation on ECM components were studied both in vivo and in vitro. Downstream targets of ALKBH5, along with their underlying mechanisms, were identified using integrated high-throughput analysis, RNA-binding protein immunoprecipitation and RNA pull-down assays. Furthermore, the therapeutic potential of exogenous ALKBH5 overexpression was evaluated in fibrotic scar models.

Results

ALKBH5 was decreased in fibroblasts derived from HTS lesions and was negatively correlated with their clinical stages. Importantly, ablation of ALKBH5 promoted the expression of COL3A1, COL1A1, and ELN, leading to pathological deposition and reconstruction of the ECM both in vivo and in vitro. From a therapeutic perspective, the exogenous overexpression of ALKBH5 significantly inhibited abnormal collagen deposition in fibrotic scar models. As determined by integrated high-throughput analysis, key ECM components including COL3A1, COL1A1, and ELN are direct downstream targets of ALKBH5. By means of its mechanism, ALKBH5 inhibits the expression of COL3A1, COL1A1, and ELN by removing m⁶A from mRNAs, thereby decreasing their stability in a YTHDF1-dependent manner.

Conclusions

Our study identified ALKBH5 as an endogenous suppressor of pathological ECM deposition, contributing to the development of a reprogrammed m6A-targeted therapy for HTSs.

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ALKBH5 介导的 m6A 去甲基化可改善皮肤病理性纤维化中细胞外基质的沉积。

背景：细胞外基质（ECM）积累增加是导致纤维化疾病发病的一个主要因素。最近的研究表明，N6-甲基腺苷（m6A）RNA 修饰在调节 RNA 稳定性方面起着关键作用，并导致各种病理状况的发生。然而，m6A 影响 ECM 沉积的确切机制仍不清楚：在这项研究中，我们以肥厚性疤痕（HTSs）为范例，研究与 ECM 相关的疾病。我们重点研究了 ALKBH5 介导的 m6A 去甲基化在肥厚性疤痕病理进展中的作用，并考察了其与临床分期的相关性。我们在体内和体外研究了 ALKBH5 消融对 ECM 成分的影响。通过综合高通量分析、RNA结合蛋白免疫沉淀和RNA牵引试验，确定了ALKBH5的下游靶点及其内在机制。此外，还在纤维化瘢痕模型中评估了外源性ALKBH5过表达的治疗潜力：结果：ALKBH5在来源于HTS病变的成纤维细胞中减少，并与临床分期呈负相关。重要的是，ALKBH5 的消减促进了 COL3A1、COL1A1 和 ELN 的表达，导致 ECM 在体内和体外的病理沉积和重建。从治疗角度来看，外源性过表达 ALKBH5 能显著抑制纤维化疤痕模型中胶原蛋白的异常沉积。通过综合高通量分析确定，包括 COL3A1、COL1A1 和 ELN 在内的关键 ECM 成分是 ALKBH5 的直接下游靶标。ALKBH5的作用机制是通过去除mRNA中的m6A，从而以依赖YTHDF1的方式降低其稳定性，从而抑制COL3A1、COL1A1和ELN的表达：我们的研究发现，ALKBH5 是病理性 ECM 沉积的内源性抑制因子，有助于开发针对 HTS 的重编程 m6A 靶向疗法。

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

Clinical and Translational Medicine Multiple-

CiteScore

15.90

自引率

1.90%

发文量

450

审稿时长

4 weeks

期刊介绍： Clinical and Translational Medicine (CTM) is an international, peer-reviewed, open-access journal dedicated to accelerating the translation of preclinical research into clinical applications and fostering communication between basic and clinical scientists. It highlights the clinical potential and application of various fields including biotechnologies, biomaterials, bioengineering, biomarkers, molecular medicine, omics science, bioinformatics, immunology, molecular imaging, drug discovery, regulation, and health policy. With a focus on the bench-to-bedside approach, CTM prioritizes studies and clinical observations that generate hypotheses relevant to patients and diseases, guiding investigations in cellular and molecular medicine. The journal encourages submissions from clinicians, researchers, policymakers, and industry professionals.