Pathophysiology of sickle cell disease is mirrored by the red blood cell metabolome

Abstract

Emerging metabolomic tools can now be used to establish metabolic signatures of specialized circulating hematopoietic cells in physiologic or pathologic conditions and in human hematologic diseases. To determine metabolomes of normal and sickle cell erythrocytes, we used an extraction method of erythrocytes metabolites coupled with a liquid chromatography-mass spectrometrybased metabolite profiling method. Comparison of these 2 metabolomes identified major changes in metabolites produced by (1) endogenous glycolysis characterized by accumulation of many glycolytic intermediates; (2) endogenous glutathione and ascorbate metabolisms characterized by accumulation of ascorbate metabolism intermediates, such as diketogulonic acid and decreased levels of both glutathione and glutathione disulfide; (3) membrane turnover, such as carnitine, or membrane transport characteristics, such as amino acids; and (4) exogenous arginine and NO metabolisms, such as spermine, spermidine, or citrulline. Finally, metabolomic analysis of young and old normal red blood cells indicates metabolites whose levels are directly related to sickle cell disease. These results show the relevance of metabolic profiling for the follow-up of sickle cell patients or other red blood cell diseases and pinpoint the importance of metabolomics to further depict the pathophysiology of human hematologic diseases. (Blood. 2011; 117(6):e57-e66) IntroductionMetabolome is defined as the complete set of metabolites present in a given biologic system that can be unicellular organism, organ, tissue, cell, or biologically relevant liquid compartments. Complementary to genomics or proteomics, the aim of metabolome analysis is to describe qualitatively and quantitatively the final products of cellular regulatory pathways and can be seen as the ultimate response of a biologic system to genetic factors and/or environmental changes. 1 Presently, metabolomics is used to describe metabolites present in simple unicellular organisms, such as Escherichia coli, 2 or in important biologic fluids, such as plasma or urine, 3,4 but the cross-talks between cells in tissues or the complex metabolism in mammalian cells make the interpretation of metabolomics data challenging in human, although important metabolites involved in pathogenesis of cancer, 5,6 diabetes, 7 and cardiovascular 8,9 and mitochondrial diseases 10 are described.Mammalian mature red blood cell (RBC) is a cell without nucleus or cytoplasmic organelles, such as mitochondria or ribosomes, easy to collect and to purify in large quantity. In human, during their 120-day life span, RBCs are perfectly adapted to oxygen, carbon dioxide, and proton transport. RBCs have also an important role in interorgan transport of metabolites, such as amino acid transport to muscles resulting from numerous amino acids receptors on RBC membranes. RBCs are also used as reporters of exogenous metabolisms as exemplified by the level of hemoglobin A1c, which is a measure of erythrocyte hemoglobin glycation and reflects mean glycemia for the...

Alterations of red blood cell metabolome in overhydrated hereditary stomatocytosis

“…haematologica | 2011; 96(12) Creatine has been reported to be a youth marker in RBCs 13 and in hemolytic anemia. Creatine concentration is expected to increase due to the short RBC lifespan (see also in sickle cell anemia 8 ). This discrepancy might be the consequence of a lower intake of creatine dependent on the sodium gradient, 14 itself altered in OHSt RBCs.…”

“…8 Control RBCs were fractioned according to the age of the RBCs. 9 The percentage of leukocytes in RBC purified samples was determined using an automated cell counter (Abacus Junior Hematology Analyzer, Diatron).…”

“…8 Here we use this strategy to distinguish metabolic changes in RBCs from OHSt patients in order to fully characterize the metabolic bases of this disease. …”

Cell Metabolomics

“…Metabolomic analysis of young and old normal red blood cells indicates metabolites whose levels are directly related to sickle cell disease (Darghouth et al, 2011). To study the metabolic changes associated with the differentiation program, the exometabolome of differentiating murine B lymphoma cells and primary B cells were compared (Garcia-Manteiga et al, 2011).…”

Nanoscale monitoring of drug actions on cell membrane using atomic force microscopy

“…When a cell becomes pathological, its morphology changes. For example, the shape of normal red blood cells is an oval biconcave disk, whereas those of anemia patients become sickle-shaped [57] . Additionally, the morphology of a normal cell changes when it transforms into a cancerous one [58] .…”