HDAC1 and 2 regulate endothelial VCAM-1 expression and atherogenesis by suppressing methylation of the GATA6 promoter.
HDAC1 and 2 regulate endothelial VCAM-1 expression and atherogenesis by suppressing methylation of the GATA6 promoter.
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HDAC1 和 2 通过抑制 GATA6 启动子甲基化来调节内皮 VCAM-1 表达和动脉粥样硬化形成
作者：
Hu C;Peng K;Wu Q;Wang Y;Fan X;Zhang DM;Passerini AG;Sun C
血管细胞粘附分子（VCAM）-1在活化的动脉内皮细胞（EC）表面上的表达增加对动脉粥样硬化的形成起关键作用，这可能部分地由表观遗传机制调节。本研究调查了临床上可用的组蛋白去乙酰化酶1和2（HDAC 1/2）抑制剂药物罗米地辛是否以及如何表观遗传地调节VCAM-1表达以抑制动脉粥样硬化。方法：在用罗米地辛处理或用HDAC 1/2靶向siRNA转染的原代人主动脉EC（HAEC）中分析VCAM-1表达。甲基化特异性PCR检测GATA 6启动子区甲基化。用染色质免疫沉淀法检测STAT 3对GATA 6启动子的富集。构建Lys 685 Arg突变以阻断STAT 3乙酰化。罗米地辛对动脉粥样硬化的潜在治疗作用在用高脂肪饮食喂养的Apoe-/-小鼠中进行评价。结果如下：罗米地辛通过同时抑制HDAC 1/2显著减弱TNFα诱导的HAEC表面VCAM-1表达和单核细胞粘附。VCAM-1的这种下调可归因于转录因子GATA 6的表达减少。罗米地辛增强STAT 3乙酰化及其与DNA甲基转移酶1（DNMT 1）的结合，导致GATA 6启动子CpG富集区+140/+255的超甲基化。在Lys 685处阻断STAT 3乙酰化破坏了DNMT 1-STAT 3相互作用，降低了GATA 6启动子甲基化，并逆转了HDAC 1/2抑制对GATA 6和VCAM-1表达的抑制作用。最后，腹膜内给予罗米地辛减少Apoe-/-小鼠中饮食诱导的动脉粥样硬化病变的发展，伴随着主动脉中GATA 6/VCAM-1表达的减少。结论：HDAC 1/2通过抑制STAT 3乙酰化依赖的GATA 6启动子甲基化参与VCAM-1表达和动脉粥样硬化。这些发现可能为HDAC 1/2靶向治疗动脉粥样硬化性心脏病提供了理论基础。
Increased expression of vascular cell adhesion molecule (VCAM)-1 on the activated arterial endothelial cell (EC) surface critically contributes to atherosclerosis which may in part be regulated by epigenetic mechanisms. This study investigated whether and how the clinically available histone deacetylases 1 and 2 (HDAC1/2) inhibitor drug Romidepsin epigenetically modulates VCAM-1 expression to suppress atherosclerosis. Methods: VCAM-1 expression was analyzed in primary human aortic EC (HAEC) treated with Romidepsin or transfected with HDAC1/2-targeting siRNA. Methylation of GATA6 promoter region was examined with methylation-specific PCR assay. Enrichment of STAT3 to GATA6 promoter was detected with chromatin immunoprecipitation. Lys685Arg mutation was constructed to block STAT3 acetylation. The potential therapeutic effect of Romidepsin on atherosclerosis was evaluated in Apoe-/- mice fed with a high-fat diet. Results: Romidepsin significantly attenuated TNFα-induced VCAM-1 expression on HAEC surface and monocyte adhesion through simultaneous inhibition of HDAC1/2. This downregulation of VCAM-1 was attributable to reduced expression of transcription factor GATA6. Romidepsin enhanced STAT3 acetylation and its binding to DNA methyltransferase 1 (DNMT1), leading to hypermethylation of the GATA6 promoter CpG-rich region at +140/+255. Blocking STAT3 acetylation at Lys685 disrupted DNMT1-STAT3 interaction, decreased GATA6 promoter methylation, and reversed the suppressive effects of HDAC1/2 inhibition on GATA6 and VCAM-1 expression. Finally, intraperitoneal administration of Romidepsin reduced diet-induced atherosclerotic lesion development in Apoe-/- mice, accompanied by a reduction in GATA6/VCAM-1 expression in the aorta. Conclusions: HDAC1/2 contributes to VCAM-1 expression and atherosclerosis by suppressing STAT3 acetylation-dependent GATA6 promoter methylation. These findings may provide a rationale for HDAC1/2-targeting therapy in atherosclerotic heart disease.