H3K27Me3 abundance is associated with a decreased barrier function of endothelial cells by directly silencing VE-cadherin expression

IF 4.9 2区医学 Q1 CARDIAC & CARDIOVASCULAR SYSTEMS

Atherosclerosis Pub Date : 2025-05-15 DOI:10.1016/j.atherosclerosis.2025.119242

Jolien Fledderus , Byambasuren Vanchin , Linda Brouwer , Timara Kuiper , Rianne M. Jongman , M. van Meurs , Martin C. Harmsen , Guido Krenning

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引用次数: 0

Abstract

Background and aims

Atherosclerosis develops mainly in predisposed, atheroprone regions characterised by the presence of disturbed flow i.e. oscillatory shear stress (OSS). OSS can induce endothelial cell (EC) activation, disruption of the EC barrier and increased permeability. The mechanisms that underly the loss of the EC barrier integrity are still incompletely understood. Enhancer of zeste homolog 2 (EZH2) and its epigenetic silencing mark H3K27Me3 are increased in the endothelium at atheroprone areas where EC barrier disruption is most prominent. Therefore, we hypothesized that increased H3K27Me3 abundance at atheroprone areas affects the barrier function of the endothelium.

Methods

A knockdown model of EZH2 in human umbilical vein EC (HUVEC) was used for RNA-seq, to identify differentially expressed genes involved in EC barrier function. Additionally, the effect of OSS on endothelial gene expression was studied by applying laminar shear stress (LSS) on y-shaped slides as a model to mimic atheroprotective and atheroprone areas in vivo. Results were corroborated by a functional study of the barrier function using trans-endothelial electric resistance (TEER).

Results

An increased H3K27Me3 abundance is present in areas under OSS, this coincides with a decreased expression of VE-cadherin. Differentially expression (DE) analysis of EZH2^KD HUVEC vs control, revealed that EZH2 regulates genes involved in cell-cell adhesion and leukocyte transmigration. Chromatin immunoprecipitation (ChIP) of H3K27Me3 showed that H3K27Me3 directly silenced CDH5 gene expression. Additionally, a reduction of EZH2 appears to increase EC barrier stability.

Conclusions

Decreased H3K27Me3 abundance in ECs is beneficial for the formation and integrity of the EC barrier.

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H3K27Me3丰度通过直接沉默VE-cadherin表达与内皮细胞屏障功能下降相关

背景和目的动脉粥样硬化主要发生在易发生动脉粥样硬化的区域，其特征是血流紊乱，即振荡剪切应力（OSS）。OSS可以诱导内皮细胞（EC）活化，破坏EC屏障，增加通透性。欧共体屏障完整性丧失的机制尚不完全清楚。zeste同源物2增强子（EZH2）及其表观遗传沉默标记H3K27Me3在动脉粥样硬化区内皮细胞中增加，这是EC屏障破坏最突出的区域。因此，我们假设动脉粥样硬化区H3K27Me3丰度的增加会影响内皮的屏障功能。方法采用人脐静脉EC (HUVEC) EZH2基因敲低模型进行rna测序，鉴定与EC屏障功能相关的差异表达基因。此外，通过在y形载玻片上施加层流剪切应力（laminar shear stress， LSS）作为模型，模拟体内动脉粥样硬化保护区和动脉粥样硬化倾向区，研究OSS对内皮基因表达的影响。使用跨内皮电阻（TEER）对屏障功能进行的功能研究证实了这一结果。结果OSS下H3K27Me3丰度升高，与VE-cadherin表达降低一致。EZH2KD HUVEC与对照组的差异表达（DE）分析显示，EZH2调控细胞-细胞粘附和白细胞转运相关基因。H3K27Me3的染色质免疫沉淀（ChIP）显示H3K27Me3直接沉默CDH5基因的表达。此外，EZH2的减少似乎增加了EC屏障的稳定性。结论EC中H3K27Me3丰度的降低有利于EC屏障的形成和完整。

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来源期刊

Atherosclerosis 医学-外周血管病

CiteScore

9.80

自引率

3.80%

发文量

1269

审稿时长

36 days

期刊介绍： Atherosclerosis has an open access mirror journal Atherosclerosis: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. Atherosclerosis brings together, from all sources, papers concerned with investigation on atherosclerosis, its risk factors and clinical manifestations. Atherosclerosis covers basic and translational, clinical and population research approaches to arterial and vascular biology and disease, as well as their risk factors including: disturbances of lipid and lipoprotein metabolism, diabetes and hypertension, thrombosis, and inflammation. The Editors are interested in original or review papers dealing with the pathogenesis, environmental, genetic and epigenetic basis, diagnosis or treatment of atherosclerosis and related diseases as well as their risk factors.