Metabolic Maturation of Human Pluripotent Stem Cell-Derived Cardiomyocytes by Inhibition of HIF1α and LDHA.
Metabolic Maturation of Human Pluripotent Stem Cell-Derived Cardiomyocytes by Inhibition of HIF1α and LDHA.
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DOI：
10.1161/circresaha.118.313249
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发表时间：
2018-10-12
影响因子：
20.1
通讯作者：
Domian IJ
中科院分区：
文献类型：
--
作者：
Hu D;Linders A;Yamak A;Correia C;Kijlstra JD;Garakani A;Xiao L;Milan DJ;van der Meer P;Serra M;Alves PM;Domian IJ
人多能干细胞衍生的心肌细胞（hPSC-CM）是一种容易获得的、稳健可重复的和生理学上合适的用于心脏疾病建模、药物发现和体外毒性筛选的人细胞来源。然而，与体内的成体心肌细胞不同，体外培养的hPSC-CM保持不成熟的代谢表型，其中大部分ATP通过有氧糖酵解而不是线粒体中的氧化磷酸化产生。关于控制hPSC-CM的代谢和功能成熟的潜在信号通路知之甚少。明确控制CM代谢途径选择的分子途径，促进CM代谢和功能成熟。我们在不同的培养基组成中培养hPSC-CM，包括含葡萄糖的培养基、补充有脂肪酸的含葡萄糖的培养基和以脂肪酸作为主要碳源的无葡萄糖培养基。我们发现，在葡萄糖存在下培养的CM主要利用有氧糖酵解，并异常上调缺氧诱导因子1-α（HIF 1 α）及其下游靶点乳酸脱氢酶A（LDHA）。相反，葡萄糖剥夺促进氧化磷酸化并抑制HIF 1 α。HIF 1 α或LDHA的小分子抑制导致有氧糖酵解转变为氧化磷酸化。同样，siRNA抑制HIF 1 α刺激氧化磷酸化，同时抑制有氧糖酵解。这种代谢转变伴随着线粒体含量和细胞ATP水平的增加。抑制HIF 1 α/LDHA可提高肌节长度和收缩力。我们发现，在标准培养条件下，HIF 1 α-LDHA轴在hPSC-CM中异常上调，阻止了它们的代谢成熟。该途径的化学或siRNA抑制导致从有氧糖酵解到氧化磷酸化的适当代谢转变。这进而改善了hPSC-CM的代谢和功能成熟。这些发现为hPSC-CM代谢的分子控制提供了关键的见解，并可用于产生生理上更成熟的CM，用于药物筛选、疾病建模和治疗目的。
Human Pluripotent Stem Cell-Derived Cardiomyocytes (hPSC-CMs) are a readily available, robustly reproducible and physiologically appropriate human cell source for cardiac disease modeling, drug discovery, and toxicity screenings in vitro. However, unlike adult myocardial cells in vivo, hPSC-CMs cultured in vitro maintain an immature metabolic phenotype where majority of ATP is produced through aerobic glycolysis instead of oxidative phosphorylation in the mitochondria. Little is known about the underlying signaling pathways controlling hPSC-CMs’ metabolic and functional maturation. To define the molecular pathways controlling CMs’ metabolic pathway selections and improve CM metabolic and functional maturation. We cultured hPSC-CMs in different media compositions including glucose-containing media, glucose-containing media supplemented with fatty acids, and glucose-free media with fatty acids as the primary carbon source. We found that CMs cultured in the presence of glucose utilized primarily aerobic glycolysis and aberrantly upregulated hypoxia-inducible factor 1-alpha (HIF1α) and its downstream target lactate dehydrogenase A (LDHA). Conversely, glucose deprivation promoted oxidative phosphorylation and repressed HIF1α. Small molecule inhibition of HIF1α or LDHA resulted in a switch from aerobic glycolysis to oxidative phosphorylation. Likewise, siRNA inhibition of HIF1α stimulated oxidative phosphorylation while inhibiting aerobic glycolysis. This metabolic shift was accompanied by an increase in mitochondrial content and cellular ATP levels. Furthermore, functional gene expressions, sarcomere length and contractility were improved by HIF1α/LDHA inhibition. We show that under standard culture conditions, the HIF1α-LDHA axis is aberrantly upregulated in hPSC-CMs, preventing their metabolic maturation. Chemical or siRNA inhibition of this pathway results in an appropriate metabolic shift from aerobic glycolysis to oxidative phosphorylation. This in turn improves metabolic and functional maturation of hPSC-CMs. These findings provide key insight into molecular control of hPSC-CMs’ metabolism and may be used to generate more physiologically mature CMs for drug screening, disease modeling and therapeutic purposes.