Biological and Genetic Determinants of Glycolysis: Phosphofructokinase Isoforms Boost Energy Status of Stored Red Blood Cells and Transfusion Outcomes

Abstract

SummaryMature red blood cells (RBCs) lack mitochondria, and thus exclusively rely on glycolysis to generate adenosine triphosphate (ATP) during aging in vivo and during storage in vitro in the blood bank. Here we identify an association between blood donor age, sex, ethnicity and end of storage levels of glycolytic metabolites in 13,029 volunteers from the Recipient Epidemiology and Donor Evaluation Study. Associations were also observed to ancestry-specific genetic polymorphisms in regions coding for phosphofructokinase 1, platelet – which we detected in mature RBCs - hexokinase 1, and for the ADP-ribosyl cyclase 1 and 2 (CD38/BST1). Gene-metabolite associations were validated in fresh and stored RBCs from 525 Diversity Outbred mice, and via multi-omics characterization of 1,929 samples from 643 RBC units during storage. ATP levels, breakdown and deamination into hypoxanthine were associated with hemolysis in vitro and in vivo, both in healthy autologous transfusion recipients and in 4,700 heterologous critically ill patients.HighlightsBlood donor age and sex affect glycolysis in stored RBCs from 13,029 volunteers;Ancestry-based genetic polymorphisms in PFKP and CD38/BST1 influence glycolysis in stored human and fresh or stored murine RBCs;PFKP is detected in pure mature RBCs and boosts glycolytic fluxes when ATP is low, such as in stored RBCs;ATP and hypoxanthine associate with RBC hemolysis in vitro and in vivo in mice and thousands of transfusion recipients.Graphical abstract