Manea Simona-Adriana, Vlad Mihaela-Loredana, Lazar Alexandra-Gela, Muresian Horia, Simionescu Maya, Manea Adrian
Institute of Cellular Biology and Pathology "Nicolae Simionescu" of the Romanian Academy, 8, B.P. Hasdeu Street, Bucharest, 050568, Romania.
Cardiovascular Surgery Department, University Hospital Bucharest, Bucharest, Romania.
J Transl Med. 2025 Mar 17;23(1):339. doi: 10.1186/s12967-025-06338-0.
Dysregulation of histone methylation-based epigenetic mechanisms leads to either transient or long-lasting transcriptomic alterations in vascular and immune cells with important consequences on atherosclerotic plaque development and stability. We hypothesized that the epigenetic enzyme SET7 lysine methyltransferase contributes to the up-regulation of NADPH oxidase (Nox) and NLRP3 inflammasome expression in atherosclerosis.
To test this hypothesis, we examined human non-atherosclerotic and atherosclerotic tissue samples, apolipoprotein E-deficient (ApoE-/-) mice, and human macrophages (Mac) employing real-time PCR, Western blot, immunofluorescence microscopy, and histological techniques. Male ApoE-/- mice with established atherosclerosis were randomized to receive concomitant with the high-fat diet, 5 mg/kg (R)-PFI-2, a selective SET7 pharmacological inhibitor, or its vehicle, every other day for 4 weeks.
The results revealed that SET7 mRNA and protein, and H3K4me1 levels were significantly elevated in human carotid atherosclerotic lesions, aorta of atherosclerotic mice, and in cultured pro-inflammatory Mac. In the atherosclerotic mice, pharmacological blockade of SET7 catalytic activity with the specific inhibitor, significantly reduced atherosclerotic plaque development, decreased the aortic up-regulation of mRNA and protein levels of Nox catalytic subunits, mitigated the formation of NT-/4HNE-protein adducts, attenuated NLRP3 gene and protein expression, and reduced pro-caspase-1 and pro-IL18 cleavage. In polarized pro-inflammatory human M1-Mac, SET7-oriented pharmacological intervention reduced the transcriptional up-regulation of Nox catalytic subunits, NLRP3, caspase-1, IL1β, and IL18, and the secretion IL1β and TNFα. Transient overexpression of SET7 in human endothelial cells enhanced mRNA levels of Nox1, Nox2, Nox4, Nox5, and p22phox.
The novel results show that SET7 regulates important mechanisms leading to enhanced formation of reactive oxygen species and pro-inflammatory cytokines release in atherosclerosis. The data recommend SET7 as a promising target for pharmacological interventions and as supportive therapeutic strategy in atherosclerotic cardiovascular diseases.
基于组蛋白甲基化的表观遗传机制失调会导致血管和免疫细胞中出现短暂或持久的转录组改变，对动脉粥样硬化斑块的发展和稳定性产生重要影响。我们推测表观遗传酶SET7赖氨酸甲基转移酶在动脉粥样硬化中促成了NADPH氧化酶（Nox）和NLRP3炎性小体表达的上调。
为验证这一假设，我们采用实时PCR、蛋白质印迹、免疫荧光显微镜和组织学技术，检测了人类非动脉粥样硬化和动脉粥样硬化组织样本、载脂蛋白E缺陷（ApoE-/-）小鼠以及人类巨噬细胞（Mac）。已患动脉粥样硬化的雄性ApoE-/-小鼠被随机分组，在高脂饮食的同时，每隔一天接受5mg/kg的（R）-PFI-2（一种选择性SET7药理学抑制剂）或其溶剂，持续4周。
结果显示，SET7 mRNA和蛋白质以及H3K4me1水平在人类颈动脉粥样硬化病变、动脉粥样硬化小鼠的主动脉以及培养的促炎性Mac中显著升高。在动脉粥样硬化小鼠中，用特异性抑制剂对SET7催化活性进行药理学阻断，可显著减少动脉粥样硬化斑块的发展，降低主动脉中Nox催化亚基mRNA和蛋白质水平的上调，减轻NT-/4HNE-蛋白质加合物的形成，减弱NLRP3基因和蛋白质表达，并减少前半胱天冬酶-1和前白细胞介素18的裂解。在极化的促炎性人类M1-Mac中，针对SET7的药理学干预降低了Nox催化亚基、NLRP3、半胱天冬酶-1、白细胞介素1β和白细胞介素18的转录上调，以及白细胞介素1β和肿瘤坏死因子α的分泌。在人类内皮细胞中短暂过表达SET7可提高Nox1、Nox2、Nox4、Nox5和p22phox的mRNA水平。
这些新结果表明，SET7调节导致动脉粥样硬化中活性氧生成增加和促炎性细胞因子释放的重要机制。这些数据表明SET7是药理学干预的一个有前景的靶点，也是动脉粥样硬化性心血管疾病的一种辅助治疗策略。
