Red cell transfusions are associated with inflammation and thrombosis, both arterial and venous, the mechanisms of which are not understood. Platelets are critically involved in hemostasis, thrombosis and inflammation. Our hypothesis is that storage-induced changes in red cells including the release of free hemoglobin and the accumulation of biologically active mediators in the supernatant have a direct deleterious effect on the recipient's platelets. We propose that unwanted effects on platelets contribute to the inflammatory and pro-thrombotic properties of red cell transfusions. We will study changes in stored leukoreduced Adsol (AS)-1 red cells and their supernatants over time, and ascertain the effects of supernatant mediators and stored red cells on recipient platelets.
In Aim 1, red cell-free hemoglobin and key lipid mediators that accumulate in microparticles and supernatant fractions of leukoreduced red cells will be defined, and we will ascertain the effects of stored red cells and these mediators on platelets. We will determine whether FDA licensed manipulations, such as washing and/or rejuvenation, ameliorate the effects of stored red cells and supernatant on platelet function. Mediators that will be studied based on our preliminary and published data include: hemoglobin which alters platelet function;F2 isoprostanes that are markers of oxidative stress and cell damage and directly affect platelet function;and red cell-derived microparticles which stimulate platelets. Assessment of biologic effects on platelets will employ aggregation, spreading and bioactive/proinflammatory mediator release. We will focus our investigations on red cell supernatant mediators because our supporting studies demonstrate that supernatant removal reduces morbidity and mortality in patients with hematologic malignancies. Our compelling preliminary laboratory data demonstrate that even leukoreduced red cells accumulate large amounts of mediators (hemoglobin, isoprostanes and microparticles) in the supernatant that alter platelet function in vitro and if transfused may alter recipient platelet function and promote vascular disease and inflammation.
In Aim 2, we will investigate the clinical importance of red cell storage time and infused mediators by correlating these with clinical and laboratory outcomes in critically ill patients. A prospective cohort study of 600 ICU patients will characterize the relationship of infused cell-free hemoglobin, isoprostanes, prostaglandins, and red cell storage time, with laboratory and clinical outcome measures of inflammation and hemostasis/thrombosis. Laboratory measures include CD40 ligand, C-reactive protein, D-dimer, blood cell count and differential. Clinical outcomes include organ failure, bleeding, thrombosis, time on ventilator and length of hospital stay. Overall, the proposed studies will determine whether the adverse effects of red cell transfusions are likely due to the stored red cells, to accumulated supernatant mediators, or to both.

Public Health Relevance

Red cell transfusions are one of the most commonly used therapies in hospitalized patients. Transfusions are associated with increased complications, including lung failure, blood clots and heart attack. Our research will discover explanations for these effects, and we will devise inexpensive strategies to reduce these complications

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL095467-04
Application #
8298227
Study Section
Special Emphasis Panel (ZHL1-CSR-S (S1))
Program Officer
Welniak, Lisbeth A
Project Start
2009-09-16
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2014-07-31
Support Year
4
Fiscal Year
2012
Total Cost
$382,388
Indirect Cost
$134,888
Name
University of Rochester
Department
Pathology
Type
Schools of Dentistry
DUNS #
041294109
City
Rochester
State
NY
Country
United States
Zip Code
14627
Lannan, Katie L; Phipps, Richard P; White, R James (2014) Thrombosis, platelets, microparticles and PAH: more than a clot. Drug Discov Today 19:1230-5
Spinelli, Sherry L; Lannan, Katie L; Casey, Ann E et al. (2014) Isoprostane and isofuran lipid mediators accumulate in stored red blood cells and influence platelet function in vitro. Transfusion 54:1569-79
Wagner, Stephen J; Glynn, Simone A; Welniak, Lisbeth A et al. (2014) Research opportunities in optimizing storage of red blood cell products. Transfusion 54:483-94
Connolly, Gregory C; Phipps, Richard P; Francis, Charles W (2014) Platelets and cancer-associated thrombosis. Semin Oncol 41:302-10
James White, R; Lannan, Katie L; Phipps, Richard P (2014) Drug discovery in pulmonary arterial hypertension: attacking the enigmatic root of a deadly weed. Drug Discov Today 19:1226-9
Refaai, Majed A; Carter, Jessie; Henrichs, Kelly F et al. (2013) Alterations of platelet function and clot formation kinetics after in vitro exposure to anti-A and -B. Transfusion 53:382-93
Lannan, Katie L; Sahler, Julie; Spinelli, Sherry L et al. (2013) Transfusion immunomodulation--the case for leukoreduced and (perhaps) washed transfusions. Blood Cells Mol Dis 50:61-8
Cholette, Jill M; Powers, Karen S; Alfieris, George M et al. (2013) Transfusion of cell saver salvaged blood in neonates and infants undergoing open heart surgery significantly reduces RBC and coagulant product transfusions and donor exposures: results of a prospective, randomized, clinical trial. Pediatr Crit Care Med 14:137-47
Rogers, Mary A M; Levine, Deborah A; Blumberg, Neil et al. (2012) Antigenic challenge in the etiology of autoimmune disease in women. J Autoimmun 38:J97-J102
Rogers, Mary A M; Levine, Deborah A; Blumberg, Neil et al. (2012) Triggers of hospitalization for venous thromboembolism. Circulation 125:2092-9

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