EIF4A3 Promotes Muscle Atrophy and Aging by Inhibiting the FAK Pathway Through NEDD9 mRNA Destabilization

IF 9.1 1区医学 Q1 GERIATRICS & GERONTOLOGY

Journal of Cachexia Sarcopenia and Muscle Pub Date : 2025-07-10 DOI:10.1002/jcsm.70010

Qian Li, Xiaohang Yin, Wensi Wan, Yi Zhou, Siqi Wang, Yuwei Yan, Jingying Chen, Xinyi Ren, Junli Gao, Yuying Chen, Yanan Zhang, Caiyue Cui, Emeli Chatterjee, Guoping Li, Ming Wu, Yan Zhang, Dongchao Lu, Tingting Yang, Yongjun Zheng, Jin Li

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引用次数: 0

Abstract

Background

Muscle atrophy has a poor prognosis, caused by various factors. Identifying a shared treatment target could address an unmet clinical need. The exon junction complex (EJC), a protein complex assembly that binds to RNA, facilitates post-transcriptional regulation by participating in mRNA splicing, mRNA export, translation and nonsense-mediated mRNA decay. This study aims to investigate the role of the EJC in muscle atrophy.

Methods

Single-cell transcriptome analysis and western blot were employed to analyse EJC expression in muscle atrophy. Overexpression of EJC helicase EIF4A3, as well as counteracting endogenous EIF4A3, was manipulated using lentiviral and adeno-associated virus 8 (AAV8) at both in vitro and in vivo levels. Imaging, RT-qPCR and immunoblot were utilized to identify phenotypes associated with muscle atrophy and aging. RNA-seq, RIP-seq, RT-qPCR and RIP-PCR were conducted to determine the targets of EIF4A3. A pharmacological approach that activates the downstream pathways in EIF4A3 knockdown muscle was employed to elucidate the molecular mechanisms of EIF4A3 in muscle atrophy.

Results

The core RNA helicase of the EJC, EIF4A3, showed increased expression in atrophied muscles and aging human muscle (+150.43%, n = 5 in young and aged human, age: 26.20 ± 6.760 vs. 73.60 ± 5.030, p < 0.001) and aged mice muscle (+74.54% in male, +61.28% in female: n = 6 in young and aged mice in male/female, age: 3 months vs. 20 months, p < 0.001). In vitro studies demonstrated that EIF4A3 overexpression promoted muscle atrophy and aging in myotubes (n = 6, p < 0.05), while EIF4A3 inhibition mitigated these effects (p < 0.05). In vivo phenotypic analysis indicated that overexpression of EIF4A3 in skeletal muscle promoted muscle atrophy (n = 10, p < 0.05) including reduced grip strength (−42.36%, p < 0.001), running capacity (−21.24%, p < 0.001), contraction force (−19.62%, p < 0.001), muscle weight (gastrocnemius muscle: −15.75%; p < 0.001; tibialis anterior muscle: −9.50%, p < 0.01), myofiber size (−11.59%, p < 0.001) and worsened molecular phenotypes (all p < 0.05). Knockdown of EIF4A3 protected against muscle atrophy induced by various stimuli, including denervation (n = 10, p < 0.05), immobilization (n = 10, p < 0.05) and angiotensin II (n = 6–10, p < 0.05) in mice. Mechanistically, Neural Precursor Cell Expressed, Developmentally Down-Regulated 9 (NEDD9) mRNA was identified as a direct target of EIF4A3. EIF4A3 promoted the decay of NEDD9 mRNA and inhibited the downstream focal adhesion kinase (FAK) and PI3K-Akt pathway, promoting muscle atrophy. Pharmacological activation of the NEDD9-FAK pathway abolished the pro-atrophy effects of EIF4A3.

Conclusions

Our findings shed significant light on the pivotal function of the EJC in muscle atrophy, revealing novel mechanisms that contribute to EJC-related disorders. Providing a target for therapeutic interventions aimed at combating muscle atrophy.

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EIF4A3通过NEDD9 mRNA失稳抑制FAK通路促进肌肉萎缩和衰老

背景：肌萎缩预后不良，其原因多种多样。确定一个共同的治疗目标可以解决未满足的临床需求。外显子连接复合体（EJC）是一种结合RNA的蛋白质复合体，通过参与mRNA剪接、mRNA输出、翻译和无义介导的mRNA衰变来促进转录后调控。本研究旨在探讨EJC在肌肉萎缩中的作用。方法采用单细胞转录组分析和western blot方法分析EJC在肌肉萎缩组织中的表达。利用慢病毒和腺相关病毒8 （AAV8）在体外和体内水平上控制EJC解旋酶EIF4A3的过表达，并抑制内源性EIF4A3。利用成像、RT-qPCR和免疫印迹技术鉴定与肌肉萎缩和衰老相关的表型。采用RNA-seq、RIP-seq、RT-qPCR和RIP-PCR检测EIF4A3的作用靶点。通过激活EIF4A3敲低肌肉下游通路的药理学方法，阐明EIF4A3在肌肉萎缩中的分子机制。结果EJC核心RNA解旋酶EIF4A3在萎缩肌肉和衰老人肌肉中（+150.43%，青年和老年人肌肉中n = 5，年龄：26.20±6.760 vs. 73.60±5.030,p < 0.001）和老年小鼠肌肉中（雄性+74.54%，雌性+61.28%；雌雄青年和老年小鼠n = 6，年龄：3个月vs. 20个月，p < 0.001）表达增加。体外研究表明，EIF4A3过表达可促进肌管肌肉萎缩和衰老（n = 6, p < 0.05），而EIF4A3抑制可减轻这些影响（p < 0.05）。体内表型分析表明，EIF4A3在骨骼肌中的过表达促进了肌肉萎缩（n = 10, p < 0.05），包括握力（- 42.36%,p < 0.001）、跑步能力（- 21.24%,p < 0.001）、收缩力（- 19.62%,p < 0.001）、肌肉重量(腓肠肌：- 15.75%；p < 0.001；胫骨前肌：- 9.50%,p < 0.01)，肌纤维大小（- 11.59%,p < 0.001）和分子表型恶化（均p <； 0.05）。敲低EIF4A3对多种刺激引起的肌肉萎缩有保护作用，包括小鼠去神经支配（n = 10, p < 0.05）、固定化（n = 10, p < 0.05）和血管紧张素II （n = 6-10, p < 0.05）。机制上，神经前体细胞表达，发育下调9 (NEDD9) mRNA被确定为EIF4A3的直接靶点。EIF4A3促进NEDD9 mRNA的衰减，抑制下游局灶黏附激酶（FAK）和PI3K-Akt通路，促进肌肉萎缩。药理激活NEDD9-FAK通路可消除EIF4A3的促萎缩作用。我们的研究结果揭示了EJC在肌肉萎缩中的关键功能，揭示了导致EJC相关疾病的新机制。提供治疗干预的目标，旨在对抗肌肉萎缩。

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

Journal of Cachexia Sarcopenia and Muscle MEDICINE, GENERAL & INTERNAL-

CiteScore

13.30

自引率

12.40%

发文量

234

审稿时长

16 weeks

期刊介绍： The Journal of Cachexia, Sarcopenia and Muscle is a peer-reviewed international journal dedicated to publishing materials related to cachexia and sarcopenia, as well as body composition and its physiological and pathophysiological changes across the lifespan and in response to various illnesses from all fields of life sciences. The journal aims to provide a reliable resource for professionals interested in related research or involved in the clinical care of affected patients, such as those suffering from AIDS, cancer, chronic heart failure, chronic lung disease, liver cirrhosis, chronic kidney failure, rheumatoid arthritis, or sepsis.