RIPK3 promotes ASIC1a-mediated fibroblast-like synoviocyte migration and invasion via malate shuttle-driven mitochondrial respiration in rheumatoid arthritis.

IF 13.3 1区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
Theranostics Pub Date : 2025-08-11 eCollection Date: 2025-01-01 DOI:10.7150/thno.113974
Weirong Hu, Ke Wang, Yalu Dong, Yucai Xu, Jing Xing, Jianzhong Zhu, Jie Ding, Yingjie Zhao, Yayun Xu, Yuanzhi Cheng, Xiaoqing Peng, Renpeng Zhou, Wei Hu, Feihu Chen
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引用次数: 0

Abstract

Objective: Synovial fibroblast migration and invasion are critical contributors to the progression of rheumatoid arthritis (RA). Acidification of local joint tissue exacerbates RA progression, but the underlying mechanisms remain unclear. This study aimed to investigate the role of acid-sensitive ion channel ASIC1a and its mediator, the RIPK3-MDH1 axis, in regulating the migration and invasion of RA fibroblast-like synoviocytes (RA-FLSs). Methods: The expression of ASIC1a, RIPK3, and MDH1 in synovial tissue from RA patients and arthritic mice was analyzed using immunofluorescence and Western blotting. RA-FLSs were stimulated with extracellular acidification (pH 6.8, mimicking local tissue conditions), and their migration and invasion were assessed via Transwell assays. The interaction between ASIC1a and RIPK3 was predicted using molecular docking and confirmed by co-immunoprecipitation (CO-IP). RIPK3-/- mice were used to establish a collagen antibody-induced arthritis (CAIA) model. Pharmacological inhibitors of ASIC1a (PcTX1) and RIPK3 (GSK-872) were employed to evaluate their therapeutic effects on migration and invasion in vitro and arthritis progression in vivo using the collagen-induced arthritis (CIA) model. Bioinformatics analyses, along with glucose, ATP, NAD+ and NADH assays, and oxygen consumption rate (OCR) measurements, were conducted to investigate the regulation of mitochondrial respiration by the RIPK3-MDH1 axis. Results: Extracellular acidification (pH 6.8) significantly enhanced the migration and invasion of RA-FLSs, effects that were abrogated by ASIC1a knockdown or pharmacological inhibition. ASIC1a activated RIPK3 through its kinase function, independent of its ion channel activity. RIPK3 activation promoted mitochondrial respiration and ATP production via MDH1-mediated malate shuttle activation. Furthermore, inhibition of the malate shuttle using Aminooxyacetic acid (Carboxymethoxylamine) hemihydrochloride (AOA) suppressed ASIC1a-mediated RA-FLSs migration and invasion. The RIPK3-MDH1 axis also maintained malate shuttle activity by enhancing glycolysis and glutamate metabolism through GLS1. Mechanistically, ASIC1a activated RIPK3, which in turn promoted MDH1-mediated malate shuttle activation, enhancing mitochondrial respiration and ATP synthesis, thereby driving RA-FLSs migration and invasion. In vivo, pharmacological inhibition of ASIC1a or RIPK3, as well as RIPK3 knockdown, significantly alleviated arthritis progression in CIA and CAIA mouse models. Conclusion: The RIPK3-MDH1 malate shuttle drives RA-FLSs migration and invasion in RA. Activation of the ASIC1a-RIPK3-MDH1 axis enhances mitochondrial respiration and ATP synthesis in RA-FLSs, highlighting this pathway as a potential therapeutic target for RA.

在类风湿关节炎中,RIPK3通过苹果酸梭驱动的线粒体呼吸促进asic1a介导的成纤维细胞样滑膜细胞的迁移和侵袭。
目的:滑膜成纤维细胞的迁移和侵袭是类风湿关节炎(RA)进展的关键因素。局部关节组织的酸化加剧了RA的进展,但潜在的机制尚不清楚。本研究旨在探讨酸敏感离子通道ASIC1a及其介质RIPK3-MDH1轴在调节RA成纤维细胞样滑膜细胞(RA- flss)迁移和侵袭中的作用。方法:采用免疫荧光法和Western blotting分析RA患者和关节炎小鼠滑膜组织中ASIC1a、RIPK3和MDH1的表达。细胞外酸化(pH 6.8,模拟局部组织条件)刺激RA-FLSs,并通过Transwell试验评估其迁移和侵袭。利用分子对接预测ASIC1a与RIPK3之间的相互作用,并通过共免疫沉淀(CO-IP)证实。采用RIPK3-/-小鼠建立胶原抗体诱导关节炎(CAIA)模型。采用胶原诱导关节炎(CIA)模型,采用ASIC1a (PcTX1)和RIPK3 (GSK-872)药物抑制剂,评估其对体外迁移和侵袭以及体内关节炎进展的治疗效果。通过生物信息学分析、葡萄糖、ATP、NAD+和NADH检测以及氧消耗率(OCR)测量,研究了RIPK3-MDH1轴对线粒体呼吸的调节。结果:细胞外酸化(pH 6.8)显著增强了RA-FLSs的迁移和侵袭,ASIC1a敲除或药物抑制可消除这种作用。ASIC1a通过其激酶功能激活RIPK3,独立于其离子通道活性。RIPK3激活通过mdh1介导的苹果酸穿梭活化促进线粒体呼吸和ATP的产生。此外,氨基乙酸(羧甲氧基胺)半盐酸盐(AOA)抑制苹果酸穿梭抑制asic1a介导的RA-FLSs迁移和侵袭。RIPK3-MDH1轴也通过GLS1增强糖酵解和谷氨酸代谢来维持苹果酸穿梭活性。机制上,ASIC1a激活RIPK3,进而促进mdh1介导的苹果酸穿梭活化,增强线粒体呼吸和ATP合成,从而驱动RA-FLSs迁移和侵袭。在体内,ASIC1a或RIPK3的药理抑制以及RIPK3的敲低可显著缓解CIA和CAIA小鼠模型的关节炎进展。结论:RIPK3-MDH1苹果酸穿梭介导RA- flss在RA中的迁移和侵袭。ASIC1a-RIPK3-MDH1轴的激活增强了RA- flss中的线粒体呼吸和ATP合成,突出了该途径作为RA的潜在治疗靶点。
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来源期刊
Theranostics
Theranostics MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
25.40
自引率
1.60%
发文量
433
审稿时长
1 months
期刊介绍: Theranostics serves as a pivotal platform for the exchange of clinical and scientific insights within the diagnostic and therapeutic molecular and nanomedicine community, along with allied professions engaged in integrating molecular imaging and therapy. As a multidisciplinary journal, Theranostics showcases innovative research articles spanning fields such as in vitro diagnostics and prognostics, in vivo molecular imaging, molecular therapeutics, image-guided therapy, biosensor technology, nanobiosensors, bioelectronics, system biology, translational medicine, point-of-care applications, and personalized medicine. Encouraging a broad spectrum of biomedical research with potential theranostic applications, the journal rigorously peer-reviews primary research, alongside publishing reviews, news, and commentary that aim to bridge the gap between the laboratory, clinic, and biotechnology industries.
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