Inflammatory gene expression profiles in Crohn's disease and ulcerative colitis: A comparative analysis using a reverse transcriptase multiplex ligation-dependent probe amplification protocol

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Abstract

Background and aims: Cytokines and their receptors play a critical role in the pathogenesis of the inflammatory bowel disease (IBD). The aim of this study was to investigate the expression profiles of inflammatory genes in inflamed and non-inflamed colonic tissue samples in patients with Crohn's disease (CD) and ulcerative colitis (UC), and to identify molecular signatures for different IBD phenotypes.

Methods: Seventy-one patients diagnosed with IBD (38 CD, 33 UC) and 15 non-IBD controls have been included in the study. For each patient, biopsy samples were obtained during colonoscopy from inflamed (L) and healthy (N) mucosa. We investigated by commercially available reverse-transcriptase multiplex ligation-dependent probe amplification (RT-MLPA) kit the mRNA expression of a set of 40 genes involved in inflammation: cytokines, chemokines, receptors, signal transduction molecules and transcription factors.

Results: In L biopsies from patients with CD, higher expression levels were found for IL-4 (p = 0.009) and IL-12p35 (p = 0.0005), whereas in L biopsy samples from patients with UC higher expression levels were found for IL-8 (p = 0.03), chemokines SCYA3 (p = 0.05), SCYA4 (p = 0.01) and glutathione S-transferase P1 (p = 0.01). In N biopsies of patients with CD higher expression levels were found for IL-1R (p = 0.01) and IL-12p35 (p = 0.007), whereas in N biopsies of patients with UC higher expression levels were found for IL-15 (p = 0.009) and SCYA8 (p = 0.001). The logistic regression analysis has indicated that low expression levels of IL-2 and IL-10, together with higher ASCA IgG titers were independently associated with penetrating/stricturing CD.

Conclusions: RT-MLPA is a sensitive and effective method for the evaluation of the profiles of inflammatory genes in IBD, with potential future applications for diagnosis, phenotypic stratification and targeted therapy.

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