title: Restoration of intracellular ATP production in banked red blood cells improves inducible ATP export and suppresses RBC-endothelial adhesion.
abstract: Transfusion of banked red blood cells (RBCs) has been associated with poor cardiovascular outcomes. Storage-induced alterations in RBC glycolytic flux, attenuated ATP export, and microvascular adhesion of transfused RBCs in vivo could contribute, but the underlying mechanisms have not been tested. We tested the novel hypothesis that improving deoxygenation-induced metabolic flux and the associated intracellular ATP generation in stored RBCs (sRBCs) results in an increased extracellular ATP export and suppresses microvascular adhesion of RBCs to endothelium in vivo following transfusion. We show deficient intracellular ATP production and ATP export by human sRBCs during deoxygenation (impairments ~42% and 49%, respectively). sRBC pretreatment with a solution containing glycolytic intermediate/purine/phosphate precursors (i.e., "PIPA") restored deoxygenation-induced intracellular ATP production and promoted extracellular ATP export (improvement ~120% and 50%, respectively). In a nude mouse model of transfusion, adhesion of human RBCs to the microvasculature in vivo was examined. Only 2% of fresh RBCs (fRBCs) transfused adhered to the vascular wall, compared with 16% of sRBCs transfused. PIPA pretreatment of sRBCs significantly reduced adhesion to just 5%. In hypoxia, adhesion of sRBCs transfused was significantly augmented (up to 21%), but not following transfusion of fRBCs or PIPA-treated sRBCs (3.5% or 6%). Enhancing the capacity for deoxygenation-induced glycolytic flux within sRBCs increases their ability to generate intracellular ATP, improves the inducible export of extracellular anti-adhesive ATP, and consequently suppresses adhesion of stored, transfused RBCs to the vascular wall in vivo.
authors.affiliation: Department of Medicine, Division of Hematology, Duke University Medical Center, Durham, North Carolina;
authors.forename: Brett S
authors.initials: BS
authors.lastname: Kirby
authors.affiliation: Optical Molecular Imaging and Analysis Core, Duke Cancer Institute, Duke University Medical Center, Durham, North Carolina;
authors.forename: Gabi
authors.initials: G
authors.lastname: Hanna
authors.affiliation: Department of Radiation Oncology, Duke University Medical Center, Durham, North Carolina;
authors.forename: Hansford C
authors.initials: HC
authors.lastname: Hendargo
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, Critical Care Medicine, Duke University Medical Center, Durham, North Carolina; and Durham Veterans Affairs Medical Center, Durham, North Carolina tim.mcmahon@duke.edu.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: McMahon
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001771:Blood Banks; D001793:Blood Preservation / Q000379:methods*; D001803:Blood Transfusion / Q000379:methods; D002448:Cell Adhesion; D015687:Cell Hypoxia; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D055806:Microvessels / Q000378:metabolism* / Q000502:physiology; D019188:Organ Preservation Solutions / Q000494:pharmacology
chemical_list: D019188:Organ Preservation Solutions; C505094:PIPA solution; D000255:Adenosine Triphosphate
keywords: adenosine triphosphate; erythrocytes; hypoxia; storage