Metabolic Profiling and Perturbations of Human CD34+ Hematopoietic Stem and Progenitors to Regulate HSPC Function

Hematopoietic stem cells (HSCs) possess self-renewal and multilineage differentiation abilities to generate blood cells and sustain hematopoiesis. Recent studies indicate that HSC metabolism is crucial for regulating their function and cell fate determination in mammals. However, a comprehensive understanding of the metabolic landscape of human HSCs across distinct developmental stages remains lacking. In this study, we performed untargeted metabolomics of Lin−CD34+ hematopoietic stem and progenitor cells (HSPCs) from human fetal liver (FL), umbilical cord blood (UCB), and adult bone marrow (aBM), revealing different developmentally associated metabolic signatures that shape HSPC function across ontogeny. Metabolomic analysis identified D-glutamine and arachidonic acid (AA) as metabolites exhibiting distinct abundance during HSPC development, suggesting their potential roles in modulating HSPC function. Transcriptomic profiling after specific metabolic treatment further revealed distinct gene expression programs associated with lineage commitment, stemness maintenance, and metabolic regulation. Functional assays demonstrated that the inhibition of glutamine metabolism with 6-diazo-5-oxo-L-norleucine (DON) induced HSPCs into quiescent cell states, improving the engraftment of HSPCs and myeloid differentiation. Conversely, exogenous AA supplementation in HSPC culture promoted proliferation and significantly enhanced megakaryocytic differentiation both in vitro and in vivo. Collectively, our study profiled the metabolic landscape of human HSPCs from embryonic to adult period through the newborn stages, suggesting that metabolic modulation could regulate HSPC function. These findings provide novel mechanistic insights and potential strategies for metabolite-based interventions to promote and enhance human HSPC function, with broad implications for basic research and regenerative medicine.