Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

IF 6.3 1区 医学 Q1 DERMATOLOGY Burns & Trauma Pub Date : 2024-10-19 DOI:10.1093/burnst/tkae024
Tianhao Li, Mingzi Zhang, Yunzhu Li, Yixin Sun, Jiuzuo Huang, Ang Zeng, Nanze Yu, Xiao Long
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Abstract

Background Keloid scarring is caused by a fibroproliferative disorder due to abnormal activation of genes, the underlying mechanism of which is still unclear. The basic helix–loop–helix transcription factor Twist-related protein 1 (TWIST1) controls cell proliferation and differentiation in tissue development and disease processes. In this study, we aimed to clarify the essential role of TWIST1 in the pathogenesis of keloids. Methods Immunohistochemistry, cell counting kit-8 assays, western blotting, PCR, matrigel invasion assays and immunofluorescence assays were applied to demonstrate the effects and mechanisms of TWIST1 in fibroblasts derived from normal skin and keloids. Mass spectrometry, ubiquitination assays, chromatin immunoprecipitation and dual luciferase reporter assay were applied to explore the interaction of TWIST1 with downstream molecules. Results In the present study, we confirmed that TWIST1 was upregulated in keloid tissue of patients and in keloid-derived fibroblasts (KFBs). In vitro, TWIST1 inhibition prevented KFB proliferation, invasion and activation. We also discovered a link between TWIST1 and the transforming growth factor β (TGF-β) signaling related molecules TGF-β receptor 1 (TΒR1), SMAD family member 2 (Smad2) and Smad3, and the fibrosis markers α-smooth muscle actin, collagen type I and collagen type III in KFBs. Mechanistically, we uncovered a brand-new mechanism by which TWIST1 interacts with myocyte enhancer factor 2A (MEF2A) and suppresses its ubiquitination and degradation. Using chromatin immunoprecipitation and dual-luciferase reporter assay, TΒR1 was identified as a novel downstream target of MEF2A, which directly binds to its promoter. Overexpression of TWIST1 promoted the recruitment of MEF2A to the TΒR1 promoter and restored TΒR1 functional expression. Conclusions Our research highlights a significant function of TWIST1 in the development of keloid and its related fibroblasts, partially facilitated by elevated MEF2A-dependent TΒR1 expression. Blocking the expression of TWIST1 in KFBs could potentially pave a novel therapeutic avenue for keloid treatment.
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扭转相关蛋白1通过调节肌细胞增强因子2A的稳定性促进瘢痕纤维母细胞中转化生长因子β受体1的形成
背景瘢痕疙瘩是由基因异常激活引起的纤维增生性疾病,其基本机制尚不清楚。基本螺旋环螺旋转录因子 Twist-related protein 1 (TWIST1) 在组织发育和疾病过程中控制着细胞的增殖和分化。本研究旨在阐明 TWIST1 在瘢痕疙瘩发病机制中的重要作用。方法 应用免疫组化、细胞计数试剂盒-8 检测、Western 印迹、PCR、matrigel 侵袭检测和免疫荧光检测来证明 TWIST1 在正常皮肤和瘢痕疙瘩成纤维细胞中的作用和机制。质谱分析、泛素化分析、染色质免疫沉淀和双荧光素酶报告分析被用来探讨 TWIST1 与下游分子的相互作用。结果 在本研究中,我们证实 TWIST1 在瘢痕疙瘩患者组织和瘢痕疙瘩衍生成纤维细胞(KFBs)中上调。在体外,抑制 TWIST1 可阻止 KFB 的增殖、侵袭和活化。我们还发现了 TWIST1 与转化生长因子 β(TGF-β)信号相关分子 TGF-β 受体 1(TΒR1)、SMAD 家族成员 2(Smad2)和 Smad3 以及 KFB 中的纤维化标志物 α-平滑肌肌动蛋白、I 型胶原和 III 型胶原之间的联系。从机理上讲,我们发现了TWIST1与肌细胞增强因子2A(MEF2A)相互作用并抑制其泛素化和降解的全新机制。通过染色质免疫沉淀和双荧光素酶报告分析,TΒR1被确定为MEF2A的一个新的下游靶点,它直接结合到MEF2A的启动子上。过表达 TWIST1 能促进 MEF2A 募集到 TΒR1 启动子并恢复 TΒR1 的功能表达。结论 我们的研究强调了 TWIST1 在瘢痕疙瘩及其相关成纤维细胞的发育过程中的重要功能,而 MEF2A 依赖性 TΒR1 表达的升高在一定程度上促进了这一功能。阻断 TWIST1 在 KFB 中的表达有可能为瘢痕疙瘩的治疗开辟一条新的治疗途径。
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来源期刊
Burns & Trauma
Burns & Trauma 医学-皮肤病学
CiteScore
8.40
自引率
9.40%
发文量
186
审稿时长
6 weeks
期刊介绍: The first open access journal in the field of burns and trauma injury in the Asia-Pacific region, Burns & Trauma publishes the latest developments in basic, clinical and translational research in the field. With a special focus on prevention, clinical treatment and basic research, the journal welcomes submissions in various aspects of biomaterials, tissue engineering, stem cells, critical care, immunobiology, skin transplantation, and the prevention and regeneration of burns and trauma injuries. With an expert Editorial Board and a team of dedicated scientific editors, the journal enjoys a large readership and is supported by Southwest Hospital, which covers authors'' article processing charges.
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