Protein disulfide isomerase A1 as a novel redox sensor in VEGFR2 signaling and angiogenesis

IF 9.2 1区 医学 Q1 PERIPHERAL VASCULAR DISEASE Angiogenesis Pub Date : 2022-08-19 DOI:10.1007/s10456-022-09852-7
Sheela Nagarkoti, Young-Mee Kim, Dipankar Ash, Archita Das, Eric Vitriol, Tracy-Ann Read, Seock-Won Youn, Varadarajan Sudhahar, Malgorzata McMenamin, Yali Hou, Harriet Boatwright, Ruth Caldwell, David W. Essex, Jaehyung Cho, Tohru Fukai, Masuko Ushio-Fukai
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引用次数: 3

Abstract

VEGFR2 signaling in endothelial cells (ECs) is regulated by reactive oxygen species (ROS) derived from NADPH oxidases (NOXs) and mitochondria, which plays an important role in postnatal angiogenesis. However, it remains unclear how highly diffusible ROS signal enhances VEGFR2 signaling and reparative angiogenesis. Protein disulfide isomerase A1 (PDIA1) functions as an oxidoreductase depending on the redox environment. We hypothesized that PDIA1 functions as a redox sensor to enhance angiogenesis. Here we showed that PDIA1 co-immunoprecipitated with VEGFR2 or colocalized with either VEGFR2 or an early endosome marker Rab5 at the perinuclear region upon stimulation of human ECs with VEGF. PDIA1 silencing significantly reduced VEGF-induced EC migration, proliferation and spheroid sprouting via inhibiting VEGFR2 signaling. Mechanistically, VEGF stimulation rapidly increased Cys-OH formation of PDIA1 via the NOX4–mitochondrial ROS axis. Overexpression of “redox-dead” mutant PDIA1 with replacement of the active four Cys residues with Ser significantly inhibited VEGF-induced PDIA1–CysOH formation and angiogenic responses via reducing VEGFR2 phosphorylation. Pdia1+/− mice showed impaired angiogenesis in developmental retina and Matrigel plug models as well as ex vivo aortic ring sprouting model. Study using hindlimb ischemia model revealed that PDIA1 expression was markedly increased in angiogenic ECs of ischemic muscles, and that ischemia-induced limb perfusion recovery and neovascularization were impaired in EC-specific Pdia1 conditional knockout mice. These results suggest that PDIA1 can sense VEGF-induced H2O2 signal via CysOH formation to promote VEGFR2 signaling and angiogenesis in ECs, thereby enhancing postnatal angiogenesis. The oxidized PDIA1 is a potential therapeutic target for treatment of ischemic vascular diseases.

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蛋白二硫化物异构酶A1作为VEGFR2信号传导和血管生成中的一种新型氧化还原传感器
内皮细胞(EC)中的VEGFR2信号传导受到来自NADPH氧化酶(NOx)和线粒体的活性氧(ROS)的调节,后者在出生后血管生成中起着重要作用。然而,目前尚不清楚高扩散性ROS信号如何增强VEGFR2信号传导和修复性血管生成。蛋白质二硫化物异构酶A1(PDIA1)根据氧化还原环境起氧化还原酶的作用。我们假设PDIA1起氧化还原传感器的作用,以增强血管生成。在这里,我们发现在用VEGF刺激人内皮细胞时,PDIA1在核周区域与VEGFR2共免疫沉淀,或与VEGF FR2或早期内体标记物Rab5共定位。PDIA1沉默通过抑制VEGFR2信号传导显著降低VEGF诱导的EC迁移、增殖和球体发芽。从机制上讲,VEGF刺激通过NOX4-线粒体ROS轴快速增加了PDIA1的Cys-OH形成。用Ser取代活性四个Cys残基的“氧化还原死亡”突变体PDIA1的过表达通过减少VEGFR2磷酸化显著抑制VEGF诱导的PDIA1–CysOH的形成和血管生成反应。Pdia1+/-小鼠在发育中的视网膜和Matrigel栓塞模型以及离体主动脉环发芽模型中表现出血管生成受损。使用后肢缺血模型的研究表明,在缺血肌肉的血管生成内皮细胞中,PDIA1的表达显著增加,并且在EC特异性PDIA1条件敲除小鼠中,缺血诱导的肢体灌注恢复和新生血管形成受损。这些结果表明,PDIA1可以通过CysOH的形成来感知VEGF诱导的H2O2信号,以促进内皮细胞中的VEGFR2信号和血管生成,从而增强出生后的血管生成。氧化的PDIA1是治疗缺血性血管疾病的潜在治疗靶点。
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来源期刊
Angiogenesis
Angiogenesis PERIPHERAL VASCULAR DISEASE-
CiteScore
21.90
自引率
8.20%
发文量
37
审稿时长
6-12 weeks
期刊介绍: Angiogenesis, a renowned international journal, seeks to publish high-quality original articles and reviews on the cellular and molecular mechanisms governing angiogenesis in both normal and pathological conditions. By serving as a primary platform for swift communication within the field of angiogenesis research, this multidisciplinary journal showcases pioneering experimental studies utilizing molecular techniques, in vitro methods, animal models, and clinical investigations into angiogenic diseases. Furthermore, Angiogenesis sheds light on cutting-edge therapeutic strategies for promoting or inhibiting angiogenesis, while also highlighting fresh markers and techniques for disease diagnosis and prognosis.
期刊最新文献
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