NRF2 Orchestrates the Metabolic Shift during Induced Pluripotent Stem Cell Reprogramming.
NRF2 Orchestrates the Metabolic Shift during Induced Pluripotent Stem Cell Reprogramming.
复制标题
DOI：
10.1016/j.celrep.2016.02.003
复制
发表时间：
2016-03-01
期刊：
影响因子：
8.8
通讯作者：
McKay TR
中科院分区：
文献类型：
--
作者：
Hawkins KE;Joy S;Delhove JM;Kotiadis VN;Fernandez E;Fitzpatrick LM;Whiteford JR;King PJ;Bolanos JP;Duchen MR;Waddington SN;McKay TR
诱导多能干细胞(IPSCs)在疾病建模和再生医学中的潜力是巨大的，但目前的方法仍然效率低下。了解iPSC重新编程背后的细胞机制，如代谢从氧化到糖酵解的能量产生，是提高其效率的关键。我们开发了一种慢病毒报告系统，通过对人皮肤成纤维细胞进行IPSC重编程，来分析活细胞中细胞信号和转录因子活性的纵向变化。我们发现，在缺氧诱导因子κ(HIFα，α)活性的短暂高峰之前，核转录因子NRFB、AP-1和NRF2的早期激活。从机制上讲，我们证明了氧化磷酸化的早期爆发和活性氧生成的增加介导了NRF2活性的增加，这反过来又启动了HIFα介导的糖酵解转变，并可能调节葡萄糖重新分配到戊糖磷酸途径。关键的是，通过Keap1过表达抑制NRF2会损害代谢重新编程，并导致IPSC克隆形成效率降低。细胞在重新编程的早期促进增殖，氧磷和ROS的产生因此在这个阶段抗氧化反应是活跃的，在HIFα激活之前，NRF2促进HIFα的激活，代谢开关，和克隆形成NRF2的激活伴随着葡萄糖的重新分配到PPP Hawkins等人。检查IPSC重新编程期间的代谢变化。他们提出，由转基因表达驱动的细胞增殖增加可以导致氧化磷酸化增加，从而产生ROS。升高的ROS激活NRF2，促进缺氧诱导因子α的激活和糖酵解的转换。
The potential of induced pluripotent stem cells (iPSCs) in disease modeling and regenerative medicine is vast, but current methodologies remain inefficient. Understanding the cellular mechanisms underlying iPSC reprogramming, such as the metabolic shift from oxidative to glycolytic energy production, is key to improving its efficiency. We have developed a lentiviral reporter system to assay longitudinal changes in cell signaling and transcription factor activity in living cells throughout iPSC reprogramming of human dermal fibroblasts. We reveal early NF-κB, AP-1, and NRF2 transcription factor activation prior to a temporal peak in hypoxia inducible factor α (HIFα) activity. Mechanistically, we show that an early burst in oxidative phosphorylation and elevated reactive oxygen species generation mediates increased NRF2 activity, which in turn initiates the HIFα-mediated glycolytic shift and may modulate glucose redistribution to the pentose phosphate pathway. Critically, inhibition of NRF2 by KEAP1 overexpression compromises metabolic reprogramming and results in reduced efficiency of iPSC colony formation. Cells increase proliferation, OXPHOS, and ROS production early in reprogramming The antioxidant response is therefore active at this stage, prior to HIFα activation NRF2 promotes HIFα activation, the metabolic switch, and colony formation NRF2 activation is concomitant with glucose redistribution to the PPP Hawkins et al. examine the metabolic shift during iPSC reprogramming. They propose that increased proliferation of cells driven by transgene expression can lead to increased oxidative phosphorylation resulting in ROS production. Elevated ROS activates NRF2, promoting HIFα activation and the switch to glycolysis.