Hypoxic activation of glucose-6-phosphate dehydrogenase controls the expression of genes involved in the pathogenesis of pulmonary hypertension through the regulation of DNA methylation
Hypoxic activation of glucose-6-phosphate dehydrogenase controls the expression of genes involved in the pathogenesis of pulmonary hypertension through the regulation of DNA methylation
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DOI：
10.1152/ajplung.00001.2020
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发表时间：
2020-04-01
影响因子：
4.9
通讯作者：
Gupte, Sachin A.
中科院分区：
文献类型：
--
作者：
Joshi, Sachindra Raj;Kitagawa, Atsushi;Gupte, Sachin A.
代谢重编程被认为在肺动脉高压（PH）中观察到的闭塞性血管病变的发病机制中是重要的。然而，将重编程代谢与基因异常表达联系起来的机制。调节PH中细胞功能表型的基因仍然是个谜。在此，我们证明，在小鼠中，缺氧诱导的PH通过葡萄糖-6-磷酸脱氢酶缺乏（G6PD（Def））来预防，并且进一步显示Cyp2c44（-/-）小鼠中建立的严重PH通过用G6PI）shRNA敲低或通过用抑制剂（N-乙基-N'-[（3 β，5 α）-17-氧代雄烷-3-基脲，NEF3）抑制G6PI）来减弱。在机制上，G6PD（D）（ef）、肺中的敲低和抑制：1）减少缺氧诱导的细胞质和线粒体代谢的变化。2)增加泰特甲基胞嘧啶双加氧酶2（Tet2）基因的表达，和3）上调编码基因和长非编码（lnc）RNA Pint的表达，其通过使转录起始位点下游的启动子侧翼区低甲基化来抑制细胞生长。这些结果表明，功能性TET2是G6PI抑制所必需的，以增加基因表达并逆转小鼠中缺氧诱导的PII。此外，G6PD活性抑制剂（NEPD）降低了代谢重编程，上调TET2和lncPINT，并分别抑制了从正常个体和特发性PAH患者的肺动脉中分离的对照和患病平滑肌细胞的生长。总的来说，这些发现证明了G6PD作为DNA甲基化调节剂的先前未被认识的功能。这些发现进一步表明，G6PD作为重编程代谢和异常基因调控之间的联系，并在调节PH发病机制中涉及的细胞表型中起着至关重要的作用，PH是一种具有高死亡率的衰弱性疾病。
Metabolic reprogramming is considered important in the pathogenesis of the occlusive vasculopathy observed in pulmonary hypertension (PH). However, the mechanisms that link reprogrammed metabolism to aberrant expression of genes. which modulate functional phenotypes of cells in PH, remain enigmatic. Herein, we demonstrate that, in mice, hypoxia-induced PH was prevented by glucose-6-phosphate dehydrogenase deficiency (G6PD(Def)), and further show that established severe PH in Cyp2c44(-/-) mice was attenuated by knockdown with G6PI) shRNA or by G6PI) inhibition with an inhibitor (N-ethyl-N'-[(3 beta,5 alpha)-17-oxoandrostan-3-yliurea, NEOU). Mechanistically, G6PD(D)(ef), knockdown and inhibition in lungs: 1) reduced hypoxia-induced changes in cytoplasmic and mitochondrial metabolism. 2) increased expression of Tet methylcytosine dioxygenase 2 (Tet2) gene, and 3) upregulated expression of the coding genes and long noncoding (lnc) RNA Pint, which inhibits cell growth, by hypomethylating the promoter flanking region downstream of the transcription start site. These results suggest functional TET2 is required for G6PI) inhibition to increase gene expression and to reverse hypoxia-induced PII in mice. Furthermore, the inhibitor of G6PD activity (NEOU) decreased metabolic reprogramming, upregulated TET2 and lncPINT, and inhibited growth of control and diseased smooth muscle cells isolated from pulmonary arteries of normal individuals and idiopathic-PAH patients, respectively. Collectively, these findings demonstrate a previously unrecognized function for G6PD as a regulator of DNA methylation. These findings further suggest that G6PD acts as a link between reprogrammed metabolism and aberrant gene regulation and plays a crucial role in regulating the phenotype of cells implicated in the pathogenesis of PH, a debilitating disorder with a high mortality rate.