2009 Vienna - European Society of Human Genetics

Complex traits and polygenic disorders
(gene: STX8; p=9.64x10-3, OR=0.34 (95%C.I.: 0.16-0.78)).
Conclusion. We replicated genetic associations for CD with 4q13.1,
UC with NKX2-3, LYRM4, STX8, both subtypes with 1q32.1.
References:
1.Rioux, J.D. et al. Nat. Genet. 39, 596-604 (2007).
2.Parkes, M. et al. Nat. Genet. 39, 830-832 (2007).
P09.057
Clinical significance of NOD2/CARD15 and TLR 4 gene SNPs in
inflammatory bowel disease.
L. Rigoli 1 , C. Romano 1 , R. A. Caruso 2 , C. Di Bella 1 , V. Procopio 1 , G. Lo Giudice
1 , M. Amorini 1 , L. Grasso 1 , P. Romeo 1 , F. Pugliatti 1 , C. Cuppari 1 , G. E. Calabrò
1 , C. Salpietro 1 , W. Fries 3 ;
1 Department of Pediatrics-Policlinico Universitario, Messina, Italy, 2 Department
of Human Pathology-Policlinico Universitario, Messina, Italy, 3 Department of
Medicine-Policlinico Universitario, Messina, Italy.
Background: To evaluate the role of genetic factors in the pathogenesis
of Crohn’s disease (CD) and ulcerative colitis (UC), we investigated
the single nucleotide polymorphisms (SNPs) of NOD2/CARD15
(R702W, G908R and L1007finsC), and Toll-like receptor 4 (TLR4)
genes (D299G and T399I) in a selected inflammatory bowel disease
(IBD) population coming from Southern Italy.
Methods: Allele and genotype frequencies of NOD2/CARD15 (R702W,
G908R and L1007finsC) and TLR4 (D299G and T399I) SNPs were
examined in 133 CD patients, in 45 UC patients, and in 103 healthy
controls.
Results: NOD2/CARD15 R702W mutation was significantly more frequent
in CD (9.8%) than in controls (2.4%, P = 0.001) and in UC (2.3%,
P = 0.03). No significant difference was found between UC patients
and control group (P > 0.05). In CD and UC patients, no significant association
with G908R variant was found L1007finsC SNP showed an
association with CD (9.8%)
compared with controls (2.9%, P = 0.002) and UC patients (2.3%, P
= 0.01). Moreover, in CD patients, G908R and L1007finsC mutations
were significantly associated with different phenotypes compared to
CD wild-type patients. No association of IBD with the TLR4 SNPs was
found in either cohort (allele frequencies: D299G-controls 3.9%, CD
3.7%, UC 3.4%, P > 0.05; T399I-controls 2.9%, CD 3.0%, UC 3.4%,
P > 0.05).
Conclusion: These findings confirm that, in our IBD patients selected
from Southern Italy, the NOD2/CARD15, but not TLR4 SNPs, are associated
with increased risk of CD.
P09.058
High resolution melting curve analysis for high-throughput sNP
genotyping in iL23R and NOD2/cARD15 genes
M. Mitrovic, U. Potocnik;
Medical faculty, Maribor, Slovenia.
Single nucleotide polymorphism (SNP) analysis is important tool in
the studies of genetic factors associated with complex diseases and
genetically influenced response to drug therapy (pharmacogenetics).
We developed a HRM method for NOD2 (rs2066845, rs2066844,
rs2066847) and IL23R (rs7517847) genes, associated with inflammatory
bowel diseases (IBD). In this study, we demonstrate, that HRM is
simple, fast and reliable method (95% confidence) for genotyping clinical
samples. HRM analysis is an efficient method for SNP detection
and/or genotyping, where homozygotes (GG and TT) were determined
with »Tm calling method« differed by 0,53°C for IL23R gene and by
0,1°C, 0,61°C in 0,45°C for NOD2gene SNPs rs2066845, rs2066844,
rs2066847, respectively. Difference between homozygotes and heterozygotes
was easily distinguishable by different melting curve shapes
with »gene scanning method«.
Additionally, we genotyped 345 Slovenian healthy controls and 295
IBD patients including 195 with Crohn’s disease (CD) and 136 with ulcerative
colitis (UC) for rs7517847 polymorphism in IL23R gene using
standard RFLP and optimized HRM methods. We found strong statistically
significant association of IL23R polymorphism with Slovenian
CD patients. Allele frequency of minor allele G was 0,46 in controls
and was reduced to 0,33 in CD patients (p