Identification of MicroRNA-124 as a Major Regulator of Enhanced Endothelial Cell Glycolysis in Pulmonary Arterial Hypertension via PTBP1 (Polypyrimidine Tract Binding Protein) and Pyruvate Kinase M2.
Identification of MicroRNA-124 as a Major Regulator of Enhanced Endothelial Cell Glycolysis in Pulmonary Arterial Hypertension via PTBP1 (Polypyrimidine Tract Binding Protein) and Pyruvate Kinase M2.
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DOI：
10.1161/circulationaha.117.028034
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发表时间：
2017-12-19
期刊：
影响因子：
37.8
通讯作者：
Morrell NW
中科院分区：
文献类型：
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作者：
Caruso P;Dunmore BJ;Schlosser K;Schoors S;Dos Santos C;Perez-Iratxeta C;Lavoie JR;Zhang H;Long L;Flockton AR;Frid MG;Upton PD;D'Alessandro A;Hadinnapola C;Kiskin FN;Taha M;Hurst LA;Ormiston ML;Hata A;Stenmark KR;Carmeliet P;Stewart DJ;Morrell NW
肺动脉高压（PAH）的特征是肺动脉内皮细胞（PAECs）的异常生长和糖酵解增强。然而，能源生产变化的潜在机制尚未确定。在这里，我们研究了由于骨形态发生蛋白受体2（BMPR 2）基因突变导致的遗传性PAH（HPAH）患者和特发性PAH（IPAH）患者的血液生长内皮细胞（BOEC）的miRNA和蛋白质组学特征，以确定异常内皮糖酵解的机制。我们假设，在PAH患者的BOEC中，使用分层系统生物学方法确定的miR-124的下调是剪接因子多嘧啶束结合蛋白（PTBP 1）表达增加的原因，导致丙酮酸激酶肌肉亚型1和2（PKM 1和2）的选择性剪接，从而增加PKM 2表达。我们质疑这种替代调节是否在PAH内皮细胞的糖酵解过度表型中起关键作用。HPAH和IPAH BOEC概括了IPAH患者PAEC中观察到的代谢异常，证实了从氧化磷酸化到有氧糖酵解的转变。miR-124的过表达或PTPB 1的siRNA沉默恢复了HPAH BOEC的正常增殖和糖酵解，纠正了糖酵解基因和乳酸产生的失调，并部分恢复了线粒体呼吸。在对照BOEC中，BMPR 2敲低降低了miR-124的表达，增加了PTPB 1，并增强了糖酵解。此外，我们观察到严重PAH的大鼠SUGEN缺氧模型中miR-124减少，PTPB 1和PKM 2表达增加以及糖酵解基因的显著失调，其特征在于BMPR 2表达减少和内皮过度增殖，支持该机制在体内的相关性。从PAH患者中分离的肺血管和循环祖细胞内皮细胞显示miR-124下调，通过PTPB 1和PKM 1/PKM 2导致PAH EC的代谢和增殖异常。因此，这种miRNA或其靶点的操作可能代表了治疗PAH的新治疗方法。
Pulmonary arterial hypertension (PAH) is characterized by abnormal growth and enhanced glycolysis of pulmonary artery endothelial cells (PAECs). However, the mechanisms underlying alterations in energy production have not been identified. Here, we examined the miRNA and proteomic profiles of blood outgrowth endothelial cells (BOECs) from patients with heritable PAH (HPAH) due to mutations in the bone morphogenetic protein receptor type 2 (BMPR2) gene and patients with idiopathic PAH (IPAH) to determine mechanisms underlying abnormal endothelial glycolysis. We hypothesized that in BOECs from PAH patients, the downregulation of miR-124, determined using a tiered systems biology approach, is responsible for increased expression of the splicing factor polypyrimidine-tract-binding protein (PTBP1), resulting in alternative splicing of pyruvate kinase muscle isoforms 1 and 2 (PKM1 and 2) and consequently, increased PKM2 expression. We questioned whether this alternative regulation plays a critical role in the hyperglycolytic phenotype of PAH endothelial cells. HPAH and IPAH BOECs recapitulated the metabolic abnormalities observed in PAECs from IPAH patients, confirming a switch from oxidative phosphorylation to aerobic glycolysis. Overexpression of miR-124, or siRNA silencing of PTPB1, restored normal proliferation and glycolysis in HPAH BOECs, corrected the dysregulation of glycolytic genes and lactate production, and partially restored mitochondrial respiration. BMPR2 knockdown in control BOECs reduced expression of miR-124, increased PTPB1, and enhanced glycolysis. Moreover, we observed reduced miR-124, increased PTPB1 and PKM2 expression and significant dysregulation of glycolytic genes in the rat SUGEN-hypoxia model of severe PAH, characterized by reduced BMPR2 expression and endothelial hyperproliferation, supporting the relevance of this mechanism in vivo. Pulmonary vascular and circulating progenitor endothelial cells isolated from patients with PAH demonstrate downregulation of miR-124 leading to the metabolic and proliferative abnormalities in PAH ECs via PTPB1 and PKM1/PKM2. Therefore, the manipulation of this miRNA, or its targets, could represent a novel therapeutic approach for the treatment of PAH.