Metabolomics-Based Elucidation of Active Metabolic Pathways in Erythrocytes and HSC-Derived Reticulocytes.
A detailed analysis of the metabolic state of human-stem-cell-derived erythrocytes allowed us to characterize the existence of active metabolic pathways in younger reticulocytes and compare them to mature erythrocytes. Using high-resolution LC-MS-based untargeted metabolomics, we found that reticulocytes had a comparatively much richer repertoire of metabolites, which spanned a range of metabolite classes. An untargeted metabolomics analysis using stable-isotope-labeled glucose showed that only glycolysis and the pentose phosphate pathway actively contributed to the biosynthesis of metabolites in erythrocytes, and these pathways were upregulated in reticulocytes. Most metabolite species found to be enriched in reticulocytes were residual pools of metabolites produced by earlier erythropoietic processes, and their systematic depletion in mature erythrocytes aligns with the simplification process, which is also seen at the cellular and the structural level. Our work shows that high-resolution LC-MS-based untargeted metabolomics provides a global coverage of the biochemical species that are present in erythrocytes. However, the incorporation of stable isotope labeling provides a more accurate description of the active metabolic processes that occur in each developmental stage. To our knowledge, this is the first detailed characterization of the active metabolic pathways of the erythroid lineage, and it provides a rich database for understanding the physiology of the maturation of reticulocytes into mature erythrocytes.
Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, Victoria 3052, Australia.
Anubhav
A
Srivastava
Walter and Eliza Hall Institute of Medical Research , Division of Infection and Immunity, Parkville, Victoria 3052, Australia.
Krystal J
KJ
Evans
Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, Victoria 3052, Australia.
Anna E
AE
Sexton
Walter and Eliza Hall Institute of Medical Research , Division of Infection and Immunity, Parkville, Victoria 3052, Australia.|Australian Institute of Tropical Health and Medicine, James Cook University , Douglas, Queensland 4814, Australia.
Louis
L
Schofield
Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University , Parkville, Victoria 3052, Australia.
Darren J
0000-0001-7497-7082
DJ
Creek
D002454:Cell Differentiation / Q000235:genetics; D002853:Chromatography, Liquid; D016208:Databases, Factual; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007553:Isotope Labeling; D050356:Lipid Metabolism / Q000235:genetics; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways / Q000235:genetics; D055432:Metabolomics; D012156:Reticulocytes / Q000166:cytology / Q000378:metabolism*; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism
D005947:Glucose
erythrocytes; metabolism; metabolomics; reticulocytes; stable-isotope labeling