School of Basic Medical Sciences (L.W., S.G., K.C., Ziling Li, Zou Li, M.S., C.W., P.C., D.F., X.F., Y.X.), Guangzhou Medical University, China.
Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital Fudan University, China (L.W.).
Hypertension. 2023 Dec;80(12):2627-2640. doi: 10.1161/HYPERTENSIONAHA.123.21382. Epub 2023 Oct 5.
Endothelial dysfunction plays a crucial role in aortic remodeling. Aerobic glycolysis and endothelial-to-mesenchymal transition (EndoMT) have, respectively, been suggested to contribute to endothelial dysfunction in many cardiovascular diseases. Here, we tested the hypothesis that glycolytic reprogramming is critical for EndoMT induction in aortic remodeling through an epigenetic mechanism mediated by a transcriptional corepressor CtBP1 (C-terminal binding protein 1), a sensor of glycolysis-derived NADH.
EndoMT program, aortic remodeling, and endothelial expression of the glycolytic activator PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase isoform 3) were evaluated in Ang (angiotensin) II-infused mice. Mice with endothelial-specific deficiency or CtBP1 inactivation, immunoprecipitation, chromatin immunoprecipitation, and luciferase reporter assay were employed to elucidate whether and how PFKFB3/CtBP1 epigenetically controls EndoMT.
The EndoMT program and increased endothelial PFKFB3 expression were induced in remodeled thoracic aortas. In TGF-β (transforming growth factor-β)-treated human endothelial cells, activated SMAD2/3 (SMAD Family Member 2/3) transcriptionally upregulated PFKFB3 expression. In turn, the TGF-β/SMAD signaling and EndoMT were compromised by silencing or inhibition of PFKFB3. Mechanistic studies revealed that PFKFB3-mediated glycolysis increased NADH content and activated the NADH-sensitive CtBP1. Through interaction with the transcription repressor E2F4 (E2F Transcription Factor 4), CtBP1 enhanced E2F4-mediated transcriptional repression of SMURF2 (SMAD ubiquitin regulatory factor 2), a negative regulator of TGF-β/SMAD2 signaling. Additionally, EC-specific deficiency or CtBP1 inactivation in mice led to attenuated Ang II-induced aortic remodeling.
Our results demonstrate a glycolysis-mediated positive feedback loop of the TGF-β signaling to induce EndoMT and indicate that therapeutically targeting endothelial PFKFB3 or CtBP1 activity could provide a basis for treating EndoMT-linked aortic remodeling.
内皮功能障碍在主动脉重塑中起着关键作用。有氧糖酵解和内皮向间充质转化（EndoMT）分别被认为是许多心血管疾病中内皮功能障碍的原因。在这里，我们通过 CtBP1（C 端结合蛋白 1）介导的表观遗传机制测试了这样一个假设，即糖酵解重编程通过 CtBP1（糖酵解衍生的 NADH 传感器）是内皮向间充质转化诱导的关键。
在血管紧张素 II 输注的小鼠中评估了 EndoMT 程序、主动脉重塑以及糖酵解激活剂 PFKFB3（6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶同工酶 3）在血管内皮中的表达。利用内皮特异性敲除或 CtBP1 失活、免疫沉淀、染色质免疫沉淀和荧光素酶报告基因检测，阐明 PFKFB3/CtBP1 是否以及如何通过表观遗传控制 EndoMT。
在重塑的胸主动脉中诱导了 EndoMT 程序和内皮 PFKFB3 表达增加。在 TGF-β（转化生长因子-β）处理的人内皮细胞中，激活的 SMAD2/3（SMAD 家族成员 2/3）转录上调 PFKFB3 表达。反过来，沉默或抑制 PFKFB3 会损害 TGF-β/SMAD 信号和 EndoMT。机制研究表明，PFKFB3 介导的糖酵解增加了 NADH 含量并激活了 NADH 敏感的 CtBP1。通过与转录抑制因子 E2F4（E2F 转录因子 4）相互作用，CtBP1 增强了 E2F4 对 TGF-β/SMAD2 信号负调节因子 SMURF2（SMAD 泛素调节因子 2）的转录抑制。此外，在小鼠中内皮特异性敲除或 CtBP1 失活导致 Ang II 诱导的主动脉重塑减弱。
我们的结果证明了 TGF-β 信号的糖酵解介导的正反馈环诱导了 EndoMT，并表明针对内皮 PFKFB3 或 CtBP1 活性的治疗方法可能为治疗与 EndoMT 相关的主动脉重塑提供基础。
