Senior Department of Hematology, the Fifth Medical Center of Chinese PLA General Hospital, Beijing, 100071, China.
Medical School of the Chinese PLA General Hospital, Beijing, 100039, China.
Stem Cell Res Ther. 2024 Sep 15;15(1):303. doi: 10.1186/s13287-024-03930-x.
Hematopoietic stem progenitor cells (HSPCs) undergo phenotypical and functional changes during their emergence and development. Although the molecular programs governing the development of human hematopoietic stem cells (HSCs) have been investigated broadly, the relationships between dynamic metabolic alterations and their functions remain poorly characterized.
In this study, we comprehensively described the proteomics of HSPCs in the human fetal liver (FL), umbilical cord blood (UCB), and adult bone marrow (aBM). The metabolic state of human HSPCs was assessed via a Seahorse assay, RT‒PCR, and flow cytometry-based metabolic-related analysis. To investigate whether perturbing glutathione metabolism affects reactive oxygen species (ROS) production, the metabolic state, and the expansion of human HSPCs, HSPCs were treated with buthionine sulfoximine (BSO), an inhibitor of glutathione synthetase, and N-acetyl-L-cysteine (NAC).
We investigated the metabolomic landscape of human HSPCs from the fetal, perinatal, and adult developmental stages by in-depth quantitative proteomics and predicted a metabolic switch from the oxidative state to the glycolytic state during human HSPC development. Seahorse assays, mitochondrial activity, ROS level, glucose uptake, and protein synthesis rate analysis supported our findings. In addition, immune-related pathways and antigen presentation were upregulated in UCB or aBM HSPCs, indicating their functional maturation upon development. Glutathione-related metabolic perturbations resulted in distinct responses in human HSPCs and progenitors. Furthermore, the molecular and immunophenotypic differences between human HSPCs at different developmental stages were revealed at the protein level for the first time.
The metabolic landscape of human HSPCs at three developmental stages (FL, UCB, and aBM), combined with proteomics and functional validations, substantially extends our understanding of HSC metabolic regulation. These findings provide valuable resources for understanding human HSC function and development during fetal and adult life.
造血干祖细胞（HSPCs）在其出现和发育过程中经历表型和功能变化。尽管已经广泛研究了调控人类造血干祖细胞（HSCs）发育的分子程序，但动态代谢改变与其功能之间的关系仍知之甚少。
在这项研究中，我们全面描述了人胎肝（FL）、脐血（UCB）和成人骨髓（aBM）中 HSPCs 的蛋白质组学。通过 Seahorse 测定、RT-PCR 和基于流式细胞术的代谢相关分析来评估人 HSPCs 的代谢状态。为了研究干扰谷胱甘肽代谢是否会影响活性氧（ROS）产生、代谢状态和人 HSPCs 的扩增，我们用谷胱甘肽合成酶抑制剂丁硫氨酸亚砜（BSO）和 N-乙酰-L-半胱氨酸（NAC）处理 HSPCs。
我们通过深入的定量蛋白质组学研究了来自胎儿、围产期和成人发育阶段的人 HSPC 的代谢组学图谱，并预测了人 HSPC 发育过程中从氧化状态到糖酵解状态的代谢转变。 Seahorse 测定、线粒体活性、ROS 水平、葡萄糖摄取和蛋白质合成速率分析支持了我们的发现。此外，UCB 或 aBM HSPCs 中免疫相关途径和抗原呈递上调，表明其在发育过程中功能成熟。谷胱甘肽相关代谢扰动导致人 HSPCs 和祖细胞产生不同的反应。此外，首次在蛋白质水平上揭示了不同发育阶段人 HSPCs 之间的分子和免疫表型差异。
三个发育阶段（FL、UCB 和 aBM）的人 HSPC 代谢图谱，结合蛋白质组学和功能验证，极大地扩展了我们对 HSC 代谢调控的理解。这些发现为理解胎儿和成人生命中人类 HSC 功能和发育提供了有价值的资源。
