Chen Liuqing, Chen Meiting, Luo Mupeng, Li Yong, Liao Bagen, Hu Min, Yu Qiuliyang
Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.
Shenzhen Key Laboratory for the Intelligent Microbial Manufacturing of Medicines, Shenzhen 518055, China.
ACS Sens. 2023 Apr 28;8(4):1518-1528. doi: 10.1021/acssensors.2c02565. Epub 2023 Mar 17.
Mapping NAD dynamics in live cells and human is essential for translating NAD interventions into effective therapies. Yet, genetically encoded NAD sensors with better specificity and pH resistance are still needed for the cost-effective monitoring of NAD in both subcellular compartments and clinical samples. Here, we introduce multicolor, resonance energy transfer-based NAD sensors covering nano- to millimolar concentration ranges for clinical NAD measurement and subcellular NAD visualization. The sensors captured the blood NAD increase induced by NMN supplementation and revealed the distinct subcellular effects of NAD precursors and modulators. The sensors then enabled high-throughput screenings for mitochondrial and nuclear NAD modulators and identified α-GPC, a cognition-related metabolite that induces NAD redistribution from mitochondria to the nucleus relative to the total adenine nucleotides, which was further confirmed by NAD FRET microscopy.
绘制活细胞和人体中的NAD动态变化对于将NAD干预转化为有效疗法至关重要。然而，为了在亚细胞区室和临床样本中经济高效地监测NAD，仍需要具有更好特异性和抗pH能力的基因编码NAD传感器。在此，我们引入了基于多色共振能量转移的NAD传感器，其涵盖了纳摩尔到毫摩尔浓度范围，用于临床NAD测量和亚细胞NAD可视化。这些传感器捕捉到了NMN补充诱导的血液NAD增加，并揭示了NAD前体和调节剂的不同亚细胞效应。然后，这些传感器能够对线粒体和细胞核NAD调节剂进行高通量筛选，并鉴定出α-GPC，一种与认知相关的代谢物，相对于总腺嘌呤核苷酸，它会诱导NAD从线粒体重新分布到细胞核，这一点通过NAD荧光共振能量转移显微镜进一步得到证实。
