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SSP1) Gene Variants in Inflammatory Bowel Disease
TLR5 SNPs investigated was found to be associated with CD or UC susceptibility. The detailed results of the genotyping analysis are shown in Table 1. Conclusion: Despite its important role as flagellin receptor, this study does not support a role for TLR5 in the genetic susceptibility of IBD. However, larger replication studies are needed to confirm this finding. Given that TLR5 may play a role in particular disease subtypes, an ongoing genotypephenotype analysis of our institution is investigating the potential effect of TLR5 variants on the IBD phenotype. Table 1
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Introduction: Osteopontin represents a multifunctional protein playing a pivotal role in chronic inflammatory and autoimmune diseases and has also found to be involved in the pathogenesis of inflammatory bowel disease (IBD). The aim of our study was to analyze the potential association and phenotypic consequences of osteopontin (OPN/SPP1) gene variants in a large and well-characterized cohort of European IBD patients. Aims & Methods: Genomic DNA from 2819 Caucasian individuals (n=841 patients with Crohn's disease (CD), n=473 patients with ulcerative colitis (UC), and n=1505 healthy unrelated controls) was analyzed for nine OPN/SPP1 SNPs (rs2728127, rs2853744, rs11730582, rs11439060, rs28357094, rs4754=p.Asp80Asp, rs1126616=p.Ala236Ala, rs1126772 and rs9138). Results: In all three subgroups (CD, UC, and controls), the allele frequencies of the nine OPN/SPP1 SNPs were in accordance with the predicted Hardy-Weinberg equilibrium. With the exception of rs4754, no significant differences in the allele frequencies were observed comparing CD and UC patients to healthy controls. Our analysis revealed a weak association of SNP rs4754 (p.Asp80Asp) with CD susceptibility (p=1.28 x 10-2; OR (95% CI) 0.85 [0.74-0.96]) and a decreased susceptibility to UC, although this association did not reach significance in univariate analysis (p=5.25 x 10-2; OR (95% CI) 0.85 [0.70-1.00]). Moreover, both associations of rs4754 (regarding CD and UC susceptibility) were not statistically significant after Bonferroni correction suggesting that these OPN/SPP1 variants are not major contributors to IBD susceptibility on their own. Conclusion: Our study could not identify osteopontin (OPN/ SPP1) gene variants as major disease susceptibility genes in CD or UC in a large European cohort. Further studies on phenotypic consequences and potential epistatic interactions will be required.
Table 1. Minor allele frequencies of TLR5 SNPs in patients with Crohn's disease (CD), ulcerative colitis (UC), and in controls. Minor allele frequencies (MAF), allelic test p-values, and odds ratios (OR, shown for the minor allele) with 95% confidence intervals (CI) are shown for the CD and UC case-control cohorts. S1195 Autism, Genetic Syndromes and Inflammatory Bowel Disease: Structural Variation and Intestinal Inflammation Gilberto Bultron, Wei Zhang, Judy H. Cho
S1199 Analysis of Non-Synonymous Single Nucleotide Polymorphism At Diamine Oxidase Gene (Refsnp ID: Rs1049793) in Patients with Crohn's Disease Natalia López-Palacios, Jose A G. Agundez, Juan Luis Mendoza, Elena Garcia-Martin, Carmen Martinez, Manuel E Fuentes, Jose M Ladero, Carlos Taxonera, Manuel DiazRubio
Introduction: Autism spectrum disorders are characterized by language impairments, social deficits, and repetitive behaviors. Recently, reports have shown that autistic patients have an increased risk of gastrointestinal inflammation and an association with copy number variation. Patients with autism have an increased risk of gastrointestinal inflammation and there have been reports of unexpected intestinal inflammation, with both low grade colitis and duodenitis with reduced disaccharides. We report copy number variation in five syndromic patients endoscopically diagnosed with intestinal inflammation and inflammatory bowel disease. Case series and methods: All five patients had a syndrome diagnosed in childhood and subsequently were diagnosed with intestinal inflammation. One child with Asperger's and Crohn's disease developed two abscesses, enterocutaneous fistula, small bowel resection, and required frequent infliximab. Another child with developmental delay and severe ulcerative colitis required prolonged intravenous steroids initially to gain remission. One patient with severe Crohn's disease and Asperger's syndrome required a subtotal colectomy and multiple steroid courses. One patient has Turner's syndrome and moderate ulcerative colitis. The final patient has pervasive developmental disorder, IgA deficiency, and a chronic noninflammatory bowel disease intestinal inflammation. Analysis of copy number variation was performed through comparative genomic hybridization using the Nimblegen 2.1 million oligonucleotide array. Results: Four of the five patients demonstrated evidence for a heterozygous duplication relative to the reference sequence on chromosome 16q23.1 in a region encompassing WWOX (WW domain-containing oxidoreductase isoform 7), which is an essential mediator of TNFalpha-induced apoptosis. Two patients demonstrated evidence for heterozygous duplications in the beta-defensin gene cluster on chromosome 8p23.1. Finally, complex patterns of duplications and deletions were observed on chromosome 17q12 in a region containing the CCL3 and CCL4 (chemokine ligands) isoforms. Conclusion: Copy number variations contributing to disease susceptibility are increasingly being defined, and a number of structural variants have been associated to autism. Intensive genomic analyses of patients with developmental delay and inflammatory bowel disease may provide novel insight into pathogenic mechanisms of these complex disorders.
The mucosa of colon and ileum of patients with Inflammatory bowel disease (IBD) presents increased numbers of mast cells and high levels of their degranulation products such as histamine and tryptase. Histamine is degraded through diamine oxidase (amiloride binding protein 1, ABP1). ABP1 enzyme activity in bowel mucosa of patients with IBD is 50% lower than in healthy individuals. The reason for this decrease remains unknown, but it may be due in part to polymorphisms in the ABP1 gene. The present study investigated whether a common single nucleotide polymorphism (SNP) (refSNP ID: rs1049793), which is located in exon 3 (C2029G) and cause amino acid substitution (His645Asp) in the ABP1 enzyme, is related to Crohn Disease (CD). AIMS: To analyse the association between the presence of a non-synonymous single nucleotide polymorphism at diamine oxidase gene and the risk of developing CD and to analyse its influence on the clinical course of these patients. METHODS: In this prospective, case-control study, 210 unrelated Caucasian consecutive CD patients were recruited at the Inflammatory Bowel Disease Unit of a single tertiary centre (Hospital Clínico San Carlos) in Madrid, Spain. All patients were phenotyped and followed up for a median time of 8.7 years (range 4.1-14.5 years). A total of 261 healthy volunteers from the same geographic area were also recruited and matched with patients. Both cases and controls were analysed for the presence of His645Asp amino acid substitutions in the diamine oxidase enzyme, using amplification-restriction procedures. The protocol was approved by the Ethics Committee of the Hospital and all patients and controls gave informed consent before inclusion in the study. RESULTS: No significant differences were found in the distribution of ABP1 alleles between CD patients and healthy volunteers for variant alleles = OR 1.15 (95% CI 0.86-1.55)]. The distribution of ABP1 genotypes did not differ when patients were subdivided according to gender, mean age at diagnosis, mean duration of the disease, family history of IBD, smoking habit, previous appendectomy or tonsillectomy, Montreal classification, perianal CD, extraintestinal manifestations of CD (cutaneous, articular, ocular and hepatic) and severity, i.e. need for immunosuppressive therapies, biological treatments and/or surgery. CONCLUSION: Our results suggest that the His645Asp polymorphism of the histamine metabolising enzyme ABP1 may not be related to the risk of developing CD. Moreover, this gene does not seem to play a role in disease activity.
S1197 Association of Peroxisome Proliferator-Activated Receptor-Gamma Pro12Ala and C161T Polymorphisms with Ulcerative Colitis in Chinese Population Umid K. Shrestha, Feng Zhou, Hongling Wang, Jin Li, Bing Xia Background and Aims Peroxisome proliferator-activated receptor-gamma (PPAR-γ) is frequently expressed in colon and PPAR-γ gene may play a role in the etiology of inflammatory bowel disease (IBD). The aims of the present study were to determine the frequency distribution of PPAR-γ Pro12Ala and C161T gene polymorphisms in Chinese patients with ulcerative colitis (UC) in central China and to explore the potential association of PPAR-γ genotypes with phenotypes of the patients. Methods One hundred seventy five Chinese patients with UC and 220 age and sex matched healthy controls were genotyped for PPAR-γ Pro12Ala and C161T polymorphisms by a method of PCR-RFLP. Results In PPAR-γ C161T polymorphism, the prevalence of T carrier was more common in UC patients than in the controls (37.1% vs. 25.5%, OR=1.73, 95% CI: 1.12~2.66, P=0.015), whereas in Pro12Ala polymorphism there was no significant association of Ala carriers with UC (10.3% vs. 10%, OR=1.61, 95%CI: 0.88-2.91, P=0.131). There were significant associations of T carrier with moderate to severe disease activities (P=0.047) and the Ala carrier with more extensive colitis (P=0.009). Conclusion The C161T polymorphism of the PPAR-γ gene was potentially associated with UC in Chinese patients in central China. Further studies are necessary to replicate the study and explore the functional implication of PPAR-γ polymorphisms in UC patients. Acknowledgment The study was supported by grants from the state public health project of China (200802156) and Hubei provincial center of clinical study for colorectal diseases (2008).
S1200 Germline Variation of a Novel NOD2/CARD15 Interacting Protein, GALNT2, Is Associated with Genetic Susceptibility to Crohn's Disease (CD) Anne M. Phillips, Johan Van Limbergen, Gail Davies, Hazel E. Drummond, Linda Smith, Amanda Smith, Jack Satsangi, Elaine Nimmo Introduction The strongest genetic determinant associated with susceptibility to CD is the NOD2/CARD15 gene. The contribution of the most common NOD2/CARD15 variants to CD in Scotland is substantially less than in other populations notably the English. The Wellcome Trust Case Control Consortium (WTCCC) has provided a wealth of geographically traceable British genotype data. We hypothesised that proteins interacting with NOD2/ CARD15 could also play a role in CD susceptibility through germline variation of the genes encoding them. Methods We performed a Yeast 2-Hybrid screen (Y2H) using NOD2/CARD15 as the bait protein, probing a cDNA library derived from the intestinal cell line, SW480. Protein interactions were confirmed in mammalian cells by co-immunoprecipitation (CoIP). One of the novel interacting proteins thus identified and confirmed in mammalian cells, was GALNT2, UDP-N-acetyl-alpha-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase 2, a catalyst of O-glycosylation, involved in the post-translational modification of mucins. WTCCC data from the GALNT2 locus were reanalysed geographically. In the Scottish cohort of 500 CD and 1000 control patients, the most significant GALNT2 SNP from the WTCCC was genotyped using Taqman. 25 additional haplotype tagging SNPs
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AGA Abstracts
AGA Abstracts
Osteopontin (OPN/SSP1) Gene Variants in Inflammatory Bowel Disease Julia Seiderer, Jürgen Glas, Helga-Paula Török, Julia Diegelmann, Burkhard Göke, Thomas Ochsenkühn, Bertram Müller-Myhsok, Stephan Brand
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Introduction: Osteopontin represents a multifunctional protein playing a pivotal role in chronic inflammatory and autoimmune diseases and has also found to be involved in the pathogenesis of inflammatory bowel disease (IBD). The aim of our study was to analyze the potential association and phenotypic consequences of osteopontin (OPN/SPP1) gene variants in a large and well-characterized cohort of European IBD patients. Aims & Methods: Genomic DNA from 2819 Caucasian individuals (n=841 patients with Crohn's disease (CD), n=473 patients with ulcerative colitis (UC), and n=1505 healthy unrelated controls) was analyzed for nine OPN/SPP1 SNPs (rs2728127, rs2853744, rs11730582, rs11439060, rs28357094, rs4754=p.Asp80Asp, rs1126616=p.Ala236Ala, rs1126772 and rs9138). Results: In all three subgroups (CD, UC, and controls), the allele frequencies of the nine OPN/SPP1 SNPs were in accordance with the predicted Hardy-Weinberg equilibrium. With the exception of rs4754, no significant differences in the allele frequencies were observed comparing CD and UC patients to healthy controls. Our analysis revealed a weak association of SNP rs4754 (p.Asp80Asp) with CD susceptibility (p=1.28 x 10-2; OR (95% CI) 0.85 [0.74-0.96]) and a decreased susceptibility to UC, although this association did not reach significance in univariate analysis (p=5.25 x 10-2; OR (95% CI) 0.85 [0.70-1.00]). Moreover, both associations of rs4754 (regarding CD and UC susceptibility) were not statistically significant after Bonferroni correction suggesting that these OPN/SPP1 variants are not major contributors to IBD susceptibility on their own. Conclusion: Our study could not identify osteopontin (OPN/ SPP1) gene variants as major disease susceptibility genes in CD or UC in a large European cohort. Further studies on phenotypic consequences and potential epistatic interactions will be required.
Table 1. Minor allele frequencies of TLR5 SNPs in patients with Crohn's disease (CD), ulcerative colitis (UC), and in controls. Minor allele frequencies (MAF), allelic test p-values, and odds ratios (OR, shown for the minor allele) with 95% confidence intervals (CI) are shown for the CD and UC case-control cohorts. S1195 Autism, Genetic Syndromes and Inflammatory Bowel Disease: Structural Variation and Intestinal Inflammation Gilberto Bultron, Wei Zhang, Judy H. Cho
S1199 Analysis of Non-Synonymous Single Nucleotide Polymorphism At Diamine Oxidase Gene (Refsnp ID: Rs1049793) in Patients with Crohn's Disease Natalia López-Palacios, Jose A G. Agundez, Juan Luis Mendoza, Elena Garcia-Martin, Carmen Martinez, Manuel E Fuentes, Jose M Ladero, Carlos Taxonera, Manuel DiazRubio
Introduction: Autism spectrum disorders are characterized by language impairments, social deficits, and repetitive behaviors. Recently, reports have shown that autistic patients have an increased risk of gastrointestinal inflammation and an association with copy number variation. Patients with autism have an increased risk of gastrointestinal inflammation and there have been reports of unexpected intestinal inflammation, with both low grade colitis and duodenitis with reduced disaccharides. We report copy number variation in five syndromic patients endoscopically diagnosed with intestinal inflammation and inflammatory bowel disease. Case series and methods: All five patients had a syndrome diagnosed in childhood and subsequently were diagnosed with intestinal inflammation. One child with Asperger's and Crohn's disease developed two abscesses, enterocutaneous fistula, small bowel resection, and required frequent infliximab. Another child with developmental delay and severe ulcerative colitis required prolonged intravenous steroids initially to gain remission. One patient with severe Crohn's disease and Asperger's syndrome required a subtotal colectomy and multiple steroid courses. One patient has Turner's syndrome and moderate ulcerative colitis. The final patient has pervasive developmental disorder, IgA deficiency, and a chronic noninflammatory bowel disease intestinal inflammation. Analysis of copy number variation was performed through comparative genomic hybridization using the Nimblegen 2.1 million oligonucleotide array. Results: Four of the five patients demonstrated evidence for a heterozygous duplication relative to the reference sequence on chromosome 16q23.1 in a region encompassing WWOX (WW domain-containing oxidoreductase isoform 7), which is an essential mediator of TNFalpha-induced apoptosis. Two patients demonstrated evidence for heterozygous duplications in the beta-defensin gene cluster on chromosome 8p23.1. Finally, complex patterns of duplications and deletions were observed on chromosome 17q12 in a region containing the CCL3 and CCL4 (chemokine ligands) isoforms. Conclusion: Copy number variations contributing to disease susceptibility are increasingly being defined, and a number of structural variants have been associated to autism. Intensive genomic analyses of patients with developmental delay and inflammatory bowel disease may provide novel insight into pathogenic mechanisms of these complex disorders.
The mucosa of colon and ileum of patients with Inflammatory bowel disease (IBD) presents increased numbers of mast cells and high levels of their degranulation products such as histamine and tryptase. Histamine is degraded through diamine oxidase (amiloride binding protein 1, ABP1). ABP1 enzyme activity in bowel mucosa of patients with IBD is 50% lower than in healthy individuals. The reason for this decrease remains unknown, but it may be due in part to polymorphisms in the ABP1 gene. The present study investigated whether a common single nucleotide polymorphism (SNP) (refSNP ID: rs1049793), which is located in exon 3 (C2029G) and cause amino acid substitution (His645Asp) in the ABP1 enzyme, is related to Crohn Disease (CD). AIMS: To analyse the association between the presence of a non-synonymous single nucleotide polymorphism at diamine oxidase gene and the risk of developing CD and to analyse its influence on the clinical course of these patients. METHODS: In this prospective, case-control study, 210 unrelated Caucasian consecutive CD patients were recruited at the Inflammatory Bowel Disease Unit of a single tertiary centre (Hospital Clínico San Carlos) in Madrid, Spain. All patients were phenotyped and followed up for a median time of 8.7 years (range 4.1-14.5 years). A total of 261 healthy volunteers from the same geographic area were also recruited and matched with patients. Both cases and controls were analysed for the presence of His645Asp amino acid substitutions in the diamine oxidase enzyme, using amplification-restriction procedures. The protocol was approved by the Ethics Committee of the Hospital and all patients and controls gave informed consent before inclusion in the study. RESULTS: No significant differences were found in the distribution of ABP1 alleles between CD patients and healthy volunteers for variant alleles = OR 1.15 (95% CI 0.86-1.55)]. The distribution of ABP1 genotypes did not differ when patients were subdivided according to gender, mean age at diagnosis, mean duration of the disease, family history of IBD, smoking habit, previous appendectomy or tonsillectomy, Montreal classification, perianal CD, extraintestinal manifestations of CD (cutaneous, articular, ocular and hepatic) and severity, i.e. need for immunosuppressive therapies, biological treatments and/or surgery. CONCLUSION: Our results suggest that the His645Asp polymorphism of the histamine metabolising enzyme ABP1 may not be related to the risk of developing CD. Moreover, this gene does not seem to play a role in disease activity.
S1197 Association of Peroxisome Proliferator-Activated Receptor-Gamma Pro12Ala and C161T Polymorphisms with Ulcerative Colitis in Chinese Population Umid K. Shrestha, Feng Zhou, Hongling Wang, Jin Li, Bing Xia Background and Aims Peroxisome proliferator-activated receptor-gamma (PPAR-γ) is frequently expressed in colon and PPAR-γ gene may play a role in the etiology of inflammatory bowel disease (IBD). The aims of the present study were to determine the frequency distribution of PPAR-γ Pro12Ala and C161T gene polymorphisms in Chinese patients with ulcerative colitis (UC) in central China and to explore the potential association of PPAR-γ genotypes with phenotypes of the patients. Methods One hundred seventy five Chinese patients with UC and 220 age and sex matched healthy controls were genotyped for PPAR-γ Pro12Ala and C161T polymorphisms by a method of PCR-RFLP. Results In PPAR-γ C161T polymorphism, the prevalence of T carrier was more common in UC patients than in the controls (37.1% vs. 25.5%, OR=1.73, 95% CI: 1.12~2.66, P=0.015), whereas in Pro12Ala polymorphism there was no significant association of Ala carriers with UC (10.3% vs. 10%, OR=1.61, 95%CI: 0.88-2.91, P=0.131). There were significant associations of T carrier with moderate to severe disease activities (P=0.047) and the Ala carrier with more extensive colitis (P=0.009). Conclusion The C161T polymorphism of the PPAR-γ gene was potentially associated with UC in Chinese patients in central China. Further studies are necessary to replicate the study and explore the functional implication of PPAR-γ polymorphisms in UC patients. Acknowledgment The study was supported by grants from the state public health project of China (200802156) and Hubei provincial center of clinical study for colorectal diseases (2008).
S1200 Germline Variation of a Novel NOD2/CARD15 Interacting Protein, GALNT2, Is Associated with Genetic Susceptibility to Crohn's Disease (CD) Anne M. Phillips, Johan Van Limbergen, Gail Davies, Hazel E. Drummond, Linda Smith, Amanda Smith, Jack Satsangi, Elaine Nimmo Introduction The strongest genetic determinant associated with susceptibility to CD is the NOD2/CARD15 gene. The contribution of the most common NOD2/CARD15 variants to CD in Scotland is substantially less than in other populations notably the English. The Wellcome Trust Case Control Consortium (WTCCC) has provided a wealth of geographically traceable British genotype data. We hypothesised that proteins interacting with NOD2/ CARD15 could also play a role in CD susceptibility through germline variation of the genes encoding them. Methods We performed a Yeast 2-Hybrid screen (Y2H) using NOD2/CARD15 as the bait protein, probing a cDNA library derived from the intestinal cell line, SW480. Protein interactions were confirmed in mammalian cells by co-immunoprecipitation (CoIP). One of the novel interacting proteins thus identified and confirmed in mammalian cells, was GALNT2, UDP-N-acetyl-alpha-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase 2, a catalyst of O-glycosylation, involved in the post-translational modification of mucins. WTCCC data from the GALNT2 locus were reanalysed geographically. In the Scottish cohort of 500 CD and 1000 control patients, the most significant GALNT2 SNP from the WTCCC was genotyped using Taqman. 25 additional haplotype tagging SNPs
A-211
AGA Abstracts
AGA Abstracts
Osteopontin (OPN/SSP1) Gene Variants in Inflammatory Bowel Disease Julia Seiderer, Jürgen Glas, Helga-Paula Török, Julia Diegelmann, Burkhard Göke, Thomas Ochsenkühn, Bertram Müller-Myhsok, Stephan Brand