Integrative analysis of metabolism, epigenetics and transcription in human haematopoietic stem cells

Haematopoietic stem cells (HSCs) experience drastic shifts in metabolic requirements to maintain the dynamic physiological demand of haematopoiesis. Thus, cellular metabolism is a fundamental aspect of HSC biology, with promising clinical applications, most of which include an ex vivo step. However, metabolic regulation of human HSCs, both in the context of clinically-relevant ex vivo cultures and in vivo in humans remains understudied. This project will utilize computational tools to integrate metabolomic, epigenetic and transcriptomic datasets in human HSCs across ex vivo and in vivo conditions, to identify key metabolites impacting HSC function. First, I will investigate how metabolic regulation shifts ex vivo, using clinically-relevant gene therapy culture conditions and a novel expansion culture system. This will identify how key metabolic pathways influence transcriptional regulation in relation to HSC self-renewal and differentiation. The role of selected metabolic pathways will then be functionally validated through perturbation experiments with small-molecule inhibitors. Finally, I will determine if these key metabolic pathways also play a role in vivo, comparing my ex vivo datasets to existing publicly available datasets of human HSCs across the lifespan. Overall, this project will build a comprehensive, multimodal map of how metabolites shape HSC function and stress response.