Travis Nemkov1, Syed M Qadri2,3,4, William P Sheffield3,4, Angelo D'Alessandro1. 1. Department of Biochemistry and Molecular Genetics, University of Colorado Denver - Anschutz Medical Campus, Aurora, CO, USA. 2. Faculty of Health Sciences, Ontario Tech University, Oshawa, ON, Canada. 3. Centre for Innovation, Canadian Blood Services, Hamilton, ON, Canada. 4. Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada.
Abstract
BACKGROUND: In response to stress, anucleate red blood cells (RBCs) can undergo a process of atypical cell death characterised by intracellular Ca2+ accumulation and phosphatidylserine (PS) externalisation. Here we studied alterations in RBC metabolism, a critical contributor to their capacity to survive environmental challenges, during this process. MATERIALS AND METHODS: Metabolomics analyses of RBCs and supernatants, using ultra-high-pressure liquid chromatography coupled to mass spectrometry, were performed after in vitro exposure of RBCs to different pathophysiological cell stressors, including starvation, extracellular hypertonicity, hyperthermia, and supraphysiological ionic stress. Cell death was examined by flow cytometry. RESULTS: Our data show that artificially enhancing RBC cytosolic Ca2+ influx significantly enhanced purine oxidation and strongly affected cellular bioenergetics by reducing glycolysis. Depleting extracellular Ca2+ curtailed starvation-induced cell death, an effect paralleled by the activation of compensatory pathways such as the pentose phosphate pathway, carboxylic acid metabolism, increased pyruvate to lactate ratios (methemoglobin reductase activation), one-carbon metabolism (protein-damage repair) and glutathione synthesis; RBCs exposed to hypertonic shock displayed a similar metabolic profile. Furthermore, cell stress promoted lipid remodelling as reflected by the levels of free fatty acids, acyl-carnitines and CoA precursors. Notably, RBC PS exposure, independently of the stressor, showed significant correlation with the levels of free fatty acids, glutamate, cystine, spermidine, tryptophan, 5-oxoproline, lactate, and hypoxanthine. DISCUSSION: In conclusion, different cell death-inducing pathophysiological stressors, encountered in various clinical conditions, result in differential RBC metabolic phenotypes, only partly explained by intracellular Ca2+ levels and ATP availability.
BACKGROUND: In response to stress, anucleate red blood cells (RBCs) can undergo a process of atypical cell death characterised by intracellular Ca2+ accumulation and phosphatidylserine (PS) externalisation. Here we studied alterations in RBC metabolism, a critical contributor to their capacity to survive environmental challenges, during this process. MATERIALS AND METHODS: Metabolomics analyses of RBCs and supernatants, using ultra-high-pressure liquid chromatography coupled to mass spectrometry, were performed after in vitro exposure of RBCs to different pathophysiological cell stressors, including starvation, extracellular hypertonicity, hyperthermia, and supraphysiological ionic stress. Cell death was examined by flow cytometry. RESULTS: Our data show that artificially enhancing RBC cytosolic Ca2+ influx significantly enhanced purine oxidation and strongly affected cellular bioenergetics by reducing glycolysis. Depleting extracellular Ca2+ curtailed starvation-induced cell death, an effect paralleled by the activation of compensatory pathways such as the pentose phosphate pathway, carboxylic acid metabolism, increased pyruvate to lactate ratios (methemoglobin reductase activation), one-carbon metabolism (protein-damage repair) and glutathione synthesis; RBCs exposed to hypertonic shock displayed a similar metabolic profile. Furthermore, cell stress promoted lipid remodelling as reflected by the levels of free fatty acids, acyl-carnitines and CoA precursors. Notably, RBC PS exposure, independently of the stressor, showed significant correlation with the levels of free fatty acids, glutamate, cystine, spermidine, tryptophan, 5-oxoproline, lactate, and hypoxanthine. DISCUSSION: In conclusion, different cell death-inducing pathophysiological stressors, encountered in various clinical conditions, result in differential RBC metabolic phenotypes, only partly explained by intracellular Ca2+ levels and ATP availability.
Authors: Angelo D'Alessandro; Alan D Gray; Zbigniew M Szczepiorkowski; Kirk Hansen; Louise H Herschel; Larry J Dumont Journal: Transfusion Date: 2017-03-10 Impact factor: 3.157
Authors: Olga Mykhailova; Carly Olafson; Tracey R Turner; Angelo DʼAlessandro; Jason P Acker Journal: Transfusion Date: 2020-08-19 Impact factor: 3.157
Authors: Kasiemobi E Pulliam; Bernadin Joseph; Amy T Makley; Charles C Caldwell; Alex B Lentsch; Michael D Goodman; Timothy A Pritts Journal: Surgery Date: 2020-08-23 Impact factor: 3.982
Authors: Rosi Bissinger; Polina Petkova-Kirova; Olga Mykhailova; Per-Arne Oldenborg; Elena Novikova; David A Donkor; Thomas Dietz; Abdulla Al Mamun Bhuyan; William P Sheffield; Marijke Grau; Ferruh Artunc; Lars Kaestner; Jason P Acker; Syed M Qadri Journal: Cell Commun Signal Date: 2020-09-18 Impact factor: 5.712
Authors: Daniel Stephenson; Travis Nemkov; Syed M Qadri; William P Sheffield; Angelo D'Alessandro Journal: Front Physiol Date: 2022-02-07 Impact factor: 4.566
Authors: Tiffany Thomas; Davide Stefanoni; Monika Dzieciatkowska; Aaron Issaian; Travis Nemkov; Ryan C Hill; Richard O Francis; Krystalyn E Hudson; Paul W Buehler; James C Zimring; Eldad A Hod; Kirk C Hansen; Steven L Spitalnik; Angelo D'Alessandro Journal: J Proteome Res Date: 2020-10-26 Impact factor: 4.466