Gene Expression Profile of Peripheral Blood Monocytes: A Step towards the Molecular Diagnosis of Celiac Disease?

<div><p>Aim</p><p>Celiac disease (CD) is a multifactorial autoimmune disease induced by ingestion of gluten in genetically predisposed individuals. Despite technological progress, the diagnosis of CD is still based on duodenal biopsy as it was 50 years ago. In this study we analysed the expression of CD-associated genes in small bowel biopsies of patients and controls in order to explore the multivariate pathway of the expression profile of CD patients. Then, using multivariant discriminant analysis, we evaluated whether the expression profiles of these genes in peripheral blood monocytes (PBMs) differed between patients and controls.</p> <p>Participants</p><p>Thirty-seven patients with active and 11 with treated CD, 40 healthy controls and 9 disease controls (Crohn’s disease patients) were enrolled.</p> <p>Results</p><p>Several genes were differentially expressed in CD patients versus controls, but the analysis of each single gene did not provided a comprehensive picture. A multivariate discriminant analysis showed that the expression of 5 genes in intestinal mucosa accounted for 93% of the difference between CD patients and controls. We then applied the same approach to PBMs, on a training set of 20 samples. The discriminant equation obtained was validated on a testing cohort of 10 additional cases and controls, and we obtained a correct classification of all CD cases and of 91% of the control samples. We applied this equation to treated CD patients and to disease controls and obtained a discrimination of 100%.</p> <p>Conclusions</p><p>The combined expression of 4 genes allows one to discriminate between CD patients and controls, and between CD patients on a gluten-free diet and disease controls. Our results contribute to the understanding of the complex interactions among CD-associated genes, and they may represent a starting point for the development of a molecular diagnosis of celiac disease.</p> </div

Distribution of the Discriminant Score of CD, Controls, Crohn and CD patients on gluten free diet.

<p>The D-score clearly separated the four groups of subjects evaluated. Only CD patients had a negative D-score. The D-score of CD patients on gluten free diet was intermediate between the scores of controls and Crohn patients.</p

mRNA expression of candidate genes in peripheral blood monocytes.

<div><p>A) <i>KIAA1109</i>expression was higher in CD (<i>p</i>=0.05) and in CD-GFD patients (<i>p</i>=0.05) than in controls, but also in Crohn peripheral monocytes (<i>p</i>=0.02); B) <i>c-REL</i> expression was lower in CD peripheral monocytes than in controls (<i>p</i><0.01), but become higher than controls (<i>p</i><0.01) and CD (<i>p</i><0.01) after one year of GFD; the same profile was observed in Crohn peripheral monocytes; C) <i>SH2B3</i> expression was lower in CD versus controls (<i>p</i>=0.04) whereas it was significantly higher in Crohn and CD-GFD patients versus controls (<i>p</i><0.01) and CD (<i>p</i><0.01). D) <i>LPP</i> expression was lower in CD peripheral monocytes than in controls (<i>p</i>=0.04); E-F) <i>TNFAIP3</i> and <i>RGS1</i> genes expression were lower in CD peripheral monocytes versus controls (<i>p</i><0.01) and higher in Crohn patients versus controls (<i>p</i><0.01) and CD patients (<i>p</i><0.01). Both genes expression levels normalized after one year of GFD;.</p> <p>RQ: relative quantification; Ctr: controls; CD: celiac disease; CD-GFD: celiac patients on a gluten-free diet; * <i>p</i><0.01, **<i>p</i><0.05.</p></div

Analysis of the levels of mRNA expression of associated genes in duodenal tissue.

<div><p>A) IL-21 is over-expressed in CD compared to controls (<i>p</i><0.01), B) IL-2 shows a very small trend of increase in CD compared to controls; C) and D) Expression of the KIAA1109 and cREL genes: the patterns are very similar and did not show any variations among the three groups.</p> <p>RQ: relative quantification; Ctr: controls; CD: celiac disease; CD-GFD: celiac patients on a gluten-free diet; * <i>p</i><0.01, **<i>p</i><0.05.</p></div

Dense genotyping identifies and localizes multiple common and rare variant association signals in celiac disease

Item does not contain fulltextUsing variants from the 1000 Genomes Project pilot European CEU dataset and data from additional resequencing studies, we densely genotyped 183 non-HLA risk loci previously associated with immune-mediated diseases in 12,041 individuals with celiac disease (cases) and 12,228 controls. We identified 13 new celiac disease risk loci reaching genome-wide significance, bringing the number of known loci (including the HLA locus) to 40. We found multiple independent association signals at over one-third of these loci, a finding that is attributable to a combination of common, low-frequency and rare genetic variants. Compared to previously available data such as those from HapMap3, our dense genotyping in a large sample collection provided a higher resolution of the pattern of linkage disequilibrium and suggested localization of many signals to finer scale regions. In particular, 29 of the 54 fine-mapped signals seemed to be localized to single genes and, in some instances, to gene regulatory elements. Altogether, we define the complex genetic architecture of the risk regions of and refine the risk signals for celiac disease, providing the next step toward uncovering the causal mechanisms of the disease