The overall goal of this project is to improve the safety and efficacy of transfusion of stored blood. There is good evidence that red blood cell storage results in loss of functionality and integrity of red blood cells over time and that this contributes to deleterious effects upon blood transfusion. This project, which is a competitive renewal of a previously funded one, is built around the hypothesis that this storage lesion is largely due to dysregulation of nitric oxide homeostasis in the blood. This disruption is due to increased nitric oxide scavenging by cell-free hemoglobin and microparticles that are released during hemolysis in stored blood and diminished nitric oxide production by the newly discovered red cell nitric oxide synthase. In this renewal we will examine the exact mechanisms of loss of NO bioavailability, as well as down-stream effects of this loss, particularly platelet activation. e will also study interactions that occur in a susceptible host receiving the transfusion. A vast array of clinical, biophysical, molecular biology, and biochemical tools will be applied to characterize the nitric oxide storage lesion in vitro in stored blood as well as in chimeric mice models, a canine model, and in human studies. In addition, therapeutics will be explored in these systems that could restore nitric oxide homeostasis by increasing nitric oxide production.

Public Health Relevance

There are several potential risks associated with transfusion of stored red blood cells, a procedure performed about 14 million times per year in patients undergoing surgery or with chronic illness. This project explores the hypothesis that deleterious effects of red cell transfusion are due to a resulting upset in the availability of the important signaling molecule nitric oxide. The project investigates the cause of this reduction in nitric oxie bioavailability and its pathological effects, and explores ways to restore nitric oxide upon red cel transfusion.

National Institute of Health (NIH)
Research Project (R01)
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Special Emphasis Panel (ZRG1)
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Welniak, Lisbeth A
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University of Pittsburgh
Internal Medicine/Medicine
Schools of Medicine
United States
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Huang, Sha; Hou, Han Wei; Kanias, Tamir et al. (2015) Towards microfluidic-based depletion of stiff and fragile human red cells that accumulate during blood storage. Lab Chip 15:448-58
Liu, Chen; Wajih, Nadeem; Liu, Xiaohua et al. (2015) Mechanisms of human erythrocytic bioactivation of nitrite. J Biol Chem 290:1281-94
Kim-Shapiro, Daniel B; Gladwin, Mark T (2015) Pitfalls in measuring NO bioavailability using NOx. Nitric Oxide 44:2-Jan
Tejero, Jesús; Gladwin, Mark T (2014) The globin superfamily: functions in nitric oxide formation and decay. Biol Chem 395:631-9
Zemke, Anna C; Shiva, Sruti; Burns, Jane L et al. (2014) Nitrite modulates bacterial antibiotic susceptibility and biofilm formation in association with airway epithelial cells. Free Radic Biol Med 77:307-16
Corti, Paola; Gladwin, Mark T (2014) Is nitrite the circulating endocrine effector of remote ischemic preconditioning? Circ Res 114:1554-7
Saraf, Santosh L; Zhang, Xu; Kanias, Tamir et al. (2014) Haemoglobinuria is associated with chronic kidney disease and its progression in patients with sickle cell anaemia. Br J Haematol 164:729-39
Wang, Dong; Cortés-Puch, Irene; Sun, Junfeng et al. (2014) Transfusion of older stored blood worsens outcomes in canines depending on the presence and severity of pneumonia. Transfusion 54:1712-24
Liu, Chen; Liu, Xiaohua; Janes, John et al. (2014) Mechanism of faster NO scavenging by older stored red blood cells. Redox Biol 2:211-9
Sparacino-Watkins, Courtney E; Tejero, Jesús; Sun, Bin et al. (2014) Nitrite reductase and nitric-oxide synthase activity of the mitochondrial molybdopterin enzymes mARC1 and mARC2. J Biol Chem 289:10345-58

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