Abstract
The aging of hematopoietic stem cells (HSCs) impairs their regenerative and self-renewal capacities, contributing to immune dysfunction and hematologic diseases, yet safe and effective rejuvenation strategies remain elusive. METHODS: This study employed a human data-driven, metabolism-centered approach to identify endogenous compounds with rejuvenating potential for aged HSCs. Genome-scale metabolic models and single-cell transcriptomic analyses were used to characterize metabolic changes in human and mouse HSCs. Aged mice were treated with cholesterol sulfate (CS), and HSC functionality was assessed via proliferation assays, DNA damage analysis, and bone marrow transplantation. Molecular docking and in vivo experiments further explored the role of the nuclear receptor RORA. RESULTS: Supplementation with CS in aged mice enhanced HSC proliferation, reduced DNA damage, and restored hematopoietic output. Metabolic modeling revealed increased CS hydrolysis as a key alteration in aged HSCs, while single-cell transcriptomics showed that CS treatment suppressed inflammatory signaling and eliminated stress-responsive HSC subpopulations. Mechanistically, CS was identified as a potent ligand that activates RORA. Functional studies suggested that RORA functions as a key mediator in this process. CONCLUSION: CS supplementation reverses aging-associated HSC decline by enhancing proliferation, reducing DNA damage, and modulating inflammatory pathways, with RORA serving as an essential mediator. These findings highlight the CS-RORA axis as a promising therapeutic target for rejuvenating aged HSCs and preserving hematopoietic function.