Systems analysis of the transcriptional response of human ileocecal epithelial cells to Clostridium difficile toxins and effects on cell cycle control

BMC Systems Biology

Erik L Hewlett¹  Jason A Papin²  Joanne A Lannigan³  Michael D Solga³  Lauren M Cave¹  Cirle A Warren¹  Glynis L Kolling¹  Mary C Gray¹  Gina M Donato¹  Kevin M D'Auria² 

[1] Division of Infectious Diseases and International Health, Department of Medicine, University of Virginia, Charlottesville, Virginia, 22908, USA;Department of Biomedical Engineering, University of Virginia, Charlottesville, Virginia, 22908, USA;Department of Microbiology, University of Virginia, Charlottesville, Virginia, 22908, USA

关键词: cell-cycle;    epithelial cell;    gene expression;    Toxin B;    Toxin A;    Clostridium difficile;   

Others  :  1144847
DOI  :  10.1186/1752-0509-6-2

 received in 2011-11-16, accepted in 2012-01-06,  发布年份 2012

【 摘 要 】

Background

Toxins A and B (TcdA and TcdB) are Clostridium difficile's principal virulence factors, yet the pathways by which they lead to inflammation and severe diarrhea remain unclear. Also, the relative role of either toxin during infection and the differences in their effects across cell lines is still poorly understood. To better understand their effects in a susceptible cell line, we analyzed the transciptome-wide gene expression response of human ileocecal epithelial cells (HCT-8) after 2, 6, and 24 hr of toxin exposure.

Results

We show that toxins elicit very similar changes in the gene expression of HCT-8 cells, with the TcdB response occurring sooner. The high similarity suggests differences between toxins are due to events beyond transcription of a single cell-type and that their relative potencies during infection may depend on differential effects across cell types within the intestine. We next performed an enrichment analysis to determine biological functions associated with changes in transcription. Differentially expressed genes were associated with response to external stimuli and apoptotic mechanisms and, at 24 hr, were predominately associated with cell-cycle control and DNA replication. To validate our systems approach, we subsequently verified a novel G₁/S and known G₂/M cell-cycle block and increased apoptosis as predicted from our enrichment analysis.

Conclusions

This study shows a successful example of a workflow deriving novel biological insight from transcriptome-wide gene expression. Importantly, we do not find any significant difference between TcdA and TcdB besides potency or kinetics. The role of each toxin in the inhibition of cell growth and proliferation, an important function of cells in the intestinal epithelium, is characterized.

【 授权许可】

   
2012 D'Auria et al; licensee BioMed Central Ltd.

【 图 表 】

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