This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Following severe trauma mononuclear cells are reprogrammed leading to alterations in innate immunity. These phenotypes are responsible for increased susceptibility to invading organisms leading to the development of organ failure. This state has been recreated in vitro by subjecting mononuclear cells to factors induced by trauma, including platelet activating factor (PAF), oxidant stress and complement 5a (C5a). Although the mechanism(s) responsible for this reprogramming remain unknown, previous work has demonstrated that this process may be associated with alterations in the protein content within specific plasma membrane microdomains that are rich in cholesterol and sphingolipids termed lipid rafts. Following injury, we hypothesize that factors induced by trauma result in the production of the lipid mediator ceramide from lipid rafts. Ceramide once produced fuses within rafts leading to the formation of macrodomains resulting in changes in membrane fluidity. Due to these changes, various proteins are recruited to the lipid raft resulting in the formation of focal adhesion-like complexes that contain some but not all of the Toll-like receptor (TLR) components. The following experimental approach will be followed: Differentiated THP-1 cells will be subjected to lipopolysaccharide (LPS) stimulation for various periods of time up to 60 min. Selected cells will be pre-treated with PAF, hydrogen peroxide or C5a for periods of time up to 30-60 min. Lipid raft protein extraction will be performed using sucrose gradient centrifugation. Harvested proteins will then be used for analysis using the LC-ESI-MS system. It is our hypothesis that assembly of these complexes and changes in lipid raft content are responsible for subsequent reprogramming that induces enhanced activation in response to subsequent infection. Based on these in vitro observations, it is our hope to then explore potential changes that occur in severely injured trauma patients in order to determine potential prognostic and therapeutic targets.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR018522-04
Application #
7359117
Study Section
Special Emphasis Panel (ZRG1-BECM (40))
Project Start
2006-07-01
Project End
2007-06-30
Budget Start
2006-07-01
Budget End
2007-06-30
Support Year
4
Fiscal Year
2006
Total Cost
$21,015
Indirect Cost
Name
Battelle Pacific Northwest Laboratories
Department
Type
DUNS #
032987476
City
Richland
State
WA
Country
United States
Zip Code
99352
Smallwood, Heather S; Duan, Susu; Morfouace, Marie et al. (2017) Targeting Metabolic Reprogramming by Influenza Infection for Therapeutic Intervention. Cell Rep 19:1640-1653
Wang, Hui; Barbieri, Christopher E; He, Jintang et al. (2017) Quantification of mutant SPOP proteins in prostate cancer using mass spectrometry-based targeted proteomics. J Transl Med 15:175
Sigdel, Tara K; Gao, Yuqian; He, Jintang et al. (2016) Mining the human urine proteome for monitoring renal transplant injury. Kidney Int 89:1244-52
Webb-Robertson, Bobbie-Jo M; Wiberg, Holli K; Matzke, Melissa M et al. (2015) Review, evaluation, and discussion of the challenges of missing value imputation for mass spectrometry-based label-free global proteomics. J Proteome Res 14:1993-2001
Ibrahim, Yehia M; Baker, Erin S; Danielson 3rd, William F et al. (2015) Development of a New Ion Mobility (Quadrupole) Time-of-Flight Mass Spectrometer. Int J Mass Spectrom 377:655-662
Ream, Thomas S; Haag, Jeremy R; Pontvianne, Frederic et al. (2015) Subunit compositions of Arabidopsis RNA polymerases I and III reveal Pol I- and Pol III-specific forms of the AC40 subunit and alternative forms of the C53 subunit. Nucleic Acids Res 43:4163-78
Malouli, Daniel; Hansen, Scott G; Nakayasu, Ernesto S et al. (2014) Cytomegalovirus pp65 limits dissemination but is dispensable for persistence. J Clin Invest 124:1928-44
Cox, Jonathan T; Marginean, Ioan; Kelly, Ryan T et al. (2014) Improving the sensitivity of mass spectrometry by using a new sheath flow electrospray emitter array at subambient pressures. J Am Soc Mass Spectrom 25:2028-37
Cao, Li; Toli?, Nikola; Qu, Yi et al. (2014) Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. Anal Biochem 452:96-102
Martin, Jessica L; Yates, Phillip A; Soysa, Radika et al. (2014) Metabolic reprogramming during purine stress in the protozoan pathogen Leishmania donovani. PLoS Pathog 10:e1003938

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