Sepsis gene expression profiling: Murine splenic compared with hepatic responses determined by using complementary DNA microarrays

ObjectiveDNA microarrays allow genome-wide assessment of changes in relative messenger RNA abundance and thus can be used to monitor changes in gene expression. The aim of this series of experiments was to gain experience in sepsis gene expression profiling in a well-accepted model of murine polymicrobial abdominal sepsis and begin characterizing (in the parlance of genomics) the sepsis “transcriptome.” DesignProspective animal study. SettingUniversity-based animal research facility. SubjectsC57BL/6 mice. InterventionsAfter induction of general anesthesia, cecal ligation and puncture were performed to induce peritonitis and polymicrobial sepsis. The control group had sham laparotomy only. Three samples of spleen and liver were collected from septic and sham animals at 24 hrs after laparotomy. Changes in expression were measured for 588 annotated mouse genes by using a commercially available complementary DNA microarray kit. Measurements and Main ResultsBroad-scale gene expression profiles were characterized for septic liver and spleen and compared with sham controls. The analytical tools used included commercially available software packages and a novel analysis program. Very little overlap was observed in the septic gene expression profiles of these two organs. Most of the genes identified have previously been linked to regulation of the inflammatory response; importantly, however, some have not. In addition, hierarchical cluster analysis showed that cecal ligation and puncture at 24 hrs induced coordinate expression of genes that alter cell signaling and survival pathways in spleen, consistent with previously published reports of sepsis-induced splenocyte apoptosis. The current limitations of microarray analysis as reflected in these studies are also discussed. ConclusionsMicroarray technology provides a powerful new tool for rapidly analyzing tissue-specific changes in gene expression induced by sepsis in animal models. To our knowledge, these data constitute the first report on the use of microarrays to determine the sepsis transcriptome in vivo.

[1]  M. Province,et al.  Functional genomics of critical illness and injury , 2002, Critical care medicine.

[2]  T G Buchman,et al.  Complex systems analysis: a tool for shock research. , 2001, Shock.

[3]  Buchman Tg,et al.  microArRAY of hope , 2001 .

[4]  K. Tracey,et al.  Mind over immunity , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  R. Hotchkiss,et al.  SEPSIS GENE EXPRESSION PROFILING II: EFFECT OF ANTIBIOTICS OVER TIME ON MURINE SPLENOCYTES AFTER CECAL LIGATION AND PUNCTURE.: 78 , 2001 .

[6]  G. Stormo,et al.  INJURY IN THE ERA OF GENOMICS , 2001, Shock.

[7]  Roger E Bumgarner,et al.  Interaction of pseudomonas aeruginosa with epithelial cells: identification of differentially regulated genes by expression microarray analysis of human cDNAs. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[8]  E. Winzeler,et al.  Genomics, gene expression and DNA arrays , 2000, Nature.

[9]  K. Tracey,et al.  Vagus nerve stimulation attenuates the systemic inflammatory response to endotoxin , 2000, Nature.

[10]  G. Sherlock Analysis of large-scale gene expression data. , 2000, Current opinion in immunology.

[11]  S. Korsmeyer,et al.  Prevention of lymphocyte cell death in sepsis improves survival in mice. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[12]  R. Hotchkiss,et al.  Inducible nitric oxide synthase (iNOS) gene deficiency increases the mortality of sepsis in mice. , 1999, Surgery.

[13]  H. Tsukagoshi,et al.  Cecal ligation and puncture peritonitis model shows decreased nicotinic acetylcholine receptor numbers in rat muscle: immunopathologic mechanisms? , 1999, Anesthesiology.

[14]  R. Hotchkiss,et al.  Apoptotic cell death in patients with sepsis, shock, and multiple organ dysfunction. , 1999, Critical care medicine.

[15]  I. Chaudry,et al.  Sex steroids regulate pro- and anti-inflammatory cytokine release by macrophages after trauma-hemorrhage. , 1999, The American journal of physiology.

[16]  Tony Pawson,et al.  Signaling Networks—Do All Roads Lead to the Same Genes? , 1999, Cell.

[17]  S. Nasraway Sepsis research: we must change course. , 1999, Critical care medicine.

[18]  B. Williams,et al.  Identification of genes differentially regulated by interferon α, β, or γ using oligonucleotide arrays , 1998 .

[19]  D. Botstein,et al.  Cluster analysis and display of genome-wide expression patterns. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[20]  J N Weinstein,et al.  Fishing Expeditions , 1998, Science.

[21]  M. Stearns,et al.  Dismissal of Faculty , 1998, Science.

[22]  R. Hotchkiss,et al.  Apoptosis in lymphoid and parenchymal cells during sepsis: findings in normal and T- and B-cell-deficient mice. , 1997, Critical care medicine.

[23]  C. Natanson,et al.  Anti-inflammatory therapies to treat sepsis and septic shock: a reassessment. , 1997, Critical care medicine.

[24]  J. Stephenson Reflecting and regrouping after failed trials, sepsis researchers forge on. , 1996, JAMA.

[25]  I. Chaudry,et al.  Evaluation of factors affecting mortality rate after sepsis in a murine cecal ligation and puncture model. , 1983, Surgery.

[26]  J. Cobb,et al.  microArRAY of hope. , 2001, Shock.

[27]  D. Botstein,et al.  Exploring the new world of the genome with DNA microarrays , 1999, Nature Genetics.

[28]  A. Meyer Death and disability from injury: a global challenge. , 1998, The Journal of trauma.

[29]  B. Williams,et al.  Identification of genes differentially regulated by interferon alpha, beta, or gamma using oligonucleotide arrays. , 1998, Proceedings of the National Academy of Sciences of the United States of America.