Roxadustat enhances inflammation and metabolic reprogramming in human leukocytes by affecting oxygen sensing

Abstract Since its approval in 2019, hypoxia-inducible factor (HIF) prolyl hydroxylase inhibitors, like roxadustat, have been used for treatment of anemia in chronic kidney disease. However, the impact of HIF stabilization on circulating leukocytes remains largely unexplored. In this study, we examined how clinically relevant concentrations of roxadustat affect human PBMCs. We evaluated the effects of roxadustat on leukocyte viability, HIF pathway activation via protein and gene expression analysis, metabolic shifts through oxygen consumption and extracellular acidification, and immune subpopulation dynamics and activation through single-cell RNA sequencing. We also explored the effects of roxadustat combined with lipopolysaccharide to simulate conditions of inflammatory hypoxia. Roxadustat did not compromise PBMC viability, but triggered HIF-1α protein accumulation, glycolytic reprogramming, and cytokine gene expression. Single-cell RNA sequencing revealed shifts in leukocyte subpopulations, and a combined treatment with lipopolysaccharide showed an enhanced inflammatory response. We found roxadustat to be a modulator of immune activity, revealing its potential to activate specific leukocyte subpopulations and amplify inflammatory responses. Our study sheds new light on the immunological dimensions of HIF stabilization and its implications for patient care, urging further exploration of its therapeutic and safety profile.