Lipids accumulate during the routine storage of blood and stimulate multiple neutrophil (PMN) functions as well as causing activation of human pulmonary microvascular endothelial cells (HMVECs). Epidemiologic studies have demonstrated that older packed red blood cells are independently associated with the development of acute lung iinjury and multiple organ failure in injured aptients. Moreover, we have implicated these lipids in human transfusion related acute lung injury (TRALI) and have proposed a two event model for this life threatening illness, similar to the pathophysiology of the acute respiratory distress syndrome. In this two event model the clinical status of the patient is the first event and the infusion of biologically active lipids in stored blood components is the second event. An animal model of TRALI has verified that the plasma and lipids from stored, but not fresh, blood and blood components cause TRALI and an in vitro model has demonstrated much of the cellular physiology required for this two event lung model. These events include the requirements of 1) endothelial activation (increase in adhesion molecules and chemokine release) resulting in PMN priming and adherence followed by 2) activation of these adherent """"""""hyperresponsive"""""""" PMNs, culminating in lung injury. Lysophosphatidylcholines (lyso-PCs) are the major biologically active species that accumulate in cellular blood components, and these compounds stimulate both PMNs and HMVECs through rapid increases in cytosolic calcium and activation of protein kinase C (PKC). We hypothesize that lipids from stored blood stimulate human PMNs and HMVECs through activation of specific PKC isoforms that stimulate endothelial beds and/or activate sequestered PMNs resulting in acute lung injury. This hypothesis will be tested through completion of the following specific aims: 1) to determine the signaling pathways of lyso-PCs from its receptor to its effector kinases; 2) to confirm the that PKC gamma is the effector kinase in lyso-PC mediated signaling; 3) to determine the role of PKC activation in PMNs and HMVECs, and 4) to investigate the effects of PKC inhibition in a well-described animal model of TRALI using a clinically tested PKC inhibitor. Completion of these specific aims will likely result in targets for in vitro, and possibly clinical, intervention to develop methods to inhibit or to attenuate the effects of these lipids and ultimately make transfusion safer.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL059355-08
Application #
7028369
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Mondoro, Traci
Project Start
1999-04-01
Project End
2008-03-31
Budget Start
2006-04-01
Budget End
2008-03-31
Support Year
8
Fiscal Year
2006
Total Cost
$293,645
Indirect Cost
Name
University of Colorado Denver
Department
Pediatrics
Type
Schools of Medicine
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045
Loi, Michele M; Kelher, Marguerite; Dzieciatkowska, Monika et al. (2018) A comparison of different methods of red blood cell leukoreduction and additive solutions on the accumulation of neutrophil-priming activity during storage. Transfusion 58:2003-2012
Silliman, Christopher C; Kelher, Marguerite R; Khan, Samina Y et al. (2017) Supernatants and lipids from stored red blood cells activate pulmonary microvascular endothelium through the BLT2 receptor and protein kinase C activation. Transfusion 57:2690-2700
Kelher, Marguerite R; McLaughlin, Nathan J D; Banerjee, Anirban et al. (2017) LysoPCs induce Hck- and PKC?-mediated activation of PKC? causing p47phox phosphorylation and membrane translocation in neutrophils. J Leukoc Biol 101:261-273
Schofield, Thomas J; Conger, Rand D; Gonzales, Joseph E et al. (2016) Harsh parenting, physical health, and the protective role of positive parent-adolescent relationships. Soc Sci Med 157:18-26
Kelher, Marguerite R; Banerjee, Anirban; Gamboni, Fabia et al. (2016) Antibodies to major histocompatibility complex class II antigens directly prime neutrophils and cause acute lung injury in a two-event in vivo rat model. Transfusion 56:3004-3011
D'Alessandro, Angelo; Nemkov, Travis; Kelher, Marguerite et al. (2015) Routine storage of red blood cell (RBC) units in additive solution-3: a comprehensive investigation of the RBC metabolome. Transfusion 55:1155-68
Silliman, Christopher C; Kelher, Marguerite R; Khan, Samina Y et al. (2014) Experimental prestorage filtration removes antibodies and decreases lipids in RBC supernatants mitigating TRALI in vivo. Blood 123:3488-95
Ball, J Bradley; Khan, Samina Y; McLaughlin, Nathan J D et al. (2012) A two-event in vitro model of acute chest syndrome: the role of secretory phospholipase A2 and neutrophils. Pediatr Blood Cancer 58:399-405
Silliman, Christopher C; Moore, Ernest E; Kelher, Marguerite R et al. (2011) Identification of lipids that accumulate during the routine storage of prestorage leukoreduced red blood cells and cause acute lung injury. Transfusion 51:2549-54
Silliman, Christopher C; Kelher, Marguerite R; Gamboni-Robertson, Fabia et al. (2010) Tumor necrosis factor-alpha causes release of cytosolic interleukin-18 from human neutrophils. Am J Physiol Cell Physiol 298:C714-24

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