Visit our Expo - Redox and Inflammation signaling 2012

Session III : Protein kinase cascades as therapeutic targets Poster III, 52
MAPKinase gene expression – as determined by microarray analysis - distinguishes
uncomplicated from complicated reconstitution after major surgical trauma
E. Marion Schneider1, Manfred Weiss2, Weidong Du1, Fabian Kriebel1, Gerhard
Leder3, Klaus Buttenschön3, Ulrich C. Liener3, Uwe Bernd Brückner3
fabiankriebel@web.de
1Sektion Experimentelle Anaesthesiologie; 2Abteilung Klinische Anaesthesiologie,
3Abteilung Chirurgie, Universitaetsklinikum Ulm, Steinhoevelstrasse 9; 89075 Ulm,
Germany
Objective: A prospective pilot study in patients with major surgical trauma at an university
hospital was set up to identify signaling pathways by microarray expression analysis, which
may be distinct in individual patients and may be indicative for complicated versus uneventful
reconstitution post trauma.
Methods: RNA was prepared from peripheral blood drawn into PAXgene tubes from three
patients before and exactly 24h after trauma. These RNA were then transcribed into cDNA,
Cy3- or Cy5-labeled, and hybridized onto a cDNA microchip, red vs. green fluorescence was
quantified using a microarray scanner. Patient (patients before and post trauma) versus control
(healthy donors) blood cell gene expressions were compared using fluorescence intensities
and appropriate normalizations.
Results: In addition to a number of rather non-specific stress response genes, activated in all
patients, we found a remarkable number of differences in gene expression patterns of
individual patients. Some of the differing genes were associated with uncomplicated
convalescence such as up-regulation of both the ERK5 pathway (MAPK7, mitogen-activated
protein kinase-7) and transcription factors which stimulate hematopoiesis and tissue
remodeling (MEF2, myocyte enhancer factor-2, BMP-2, bone morphogenic protein-2,
TNFRSF11a (TNF-R superfamily, syn. RANK), and RUNX-1, runt related transcription
factor-1). Chemokine genes active in stem cell recruitment from the bone marrow as well as
dendritic cell and NK maturation (SCYA14 (HCC-1, hemofiltrate cc chemokine )), were
increased as well as activators of the lymphoid compartment (TNFRSF7 (CD27), CD3zeta
and perforin (PRF1)). In contrast, all these transcripts were down-regulated in complicated
reconstitution and later development of septic shock. Moreover, p38 kinase (MAPK14), S100
molecules and members of the lipoxygenase pathway were associated with a more eventful
outcome. In none of the patients, we found c-Jun-stress activated factor (JNK) pathway to be
markedly activated, also p105 (Rel) or the “NFkB inducible kinase” (NIK, MAP3K14) were
not found to be up-regulated post surgical trauma.
Conclusions: Microarray expression studies are a promising tool for screening and then
selecting differentially regulated genes in favorable as compared to complicated reconstitution
post trauma. Gene up-regulation patterns may emphasize previously unrecognized molecules
and signaling pathways encouraging the appropriate protein and functional assays in an
individual patient but also in larger patient cohorts.
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