M2082 Association Between Toll-Like 4, Cd14 Polymorphisms and Inflammatory Bowel Disease in Turkish...
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14% and 34%, respectively. Our aim in this study was to determine the allele frequencies of the two disease-causing polymorphisms in a Swedish cohort of patients with CD. Methods: Blood samples were collected from 125 consecutive CD patients (64 men, 61 women). The samples were frozen and stored at -70 centigrade. DNA was isolated and amplified using specific primers and the Q705K polymorphism of CIAS1 and C10X polymorphism of CARD8 were detected by the commercially available Megabace SNuPe genotyping kit. Moreover, the three most common polymorphisms of NOD2 were assessed, i.e. insC, 702, and 908, respectively. The results were compared to blood DNA bank controls (n=806). Results: At least one allele variant in the NOD2 gene was found in 16 % of CD patients, OR=2.32 (95 % CI: 1.39-3.88). Ten percent of the patients carried heterozygote polymorphisms in both of the CIAS1 and CARD8 genes, OR=2.34 (95 % CI: 1.13-4.84). None of these patients had polymorphisms in the NOD2 gene. Conclusions: Compound polymorphisms of CIAS1 and CARD8 doubled the susceptibility for CD in our cohort. This may be an important genetic risk factor in individuals with normal NOD2. However, the data needs confirmation in larger patient groups. M2081 Glucocorticoid Receptor Polymorphism in Inflammatory Bowel Disease Emese Mihály, Péter Gergics, Pál Miheller, Károly Rácz, Zsolt Tulassay Background and aims: Glucocorticoids are used to treat a wide variety of inflammatory diseases. They are potent inhibitors of T cell activation and proinflammatory cytokines and effectively used to treat active inflammatory bowel disease (IBD). However, there is large variability in responsiveness and sensitivity between individuals to glucocorticoid therapy, both with regard to efficacy and the prevalence and severity of the side effects. Single nucleotide gene polymorphism of the glucocorticoid receptor results in diverse clinical manifestations. Most of the mutations were found associated with glucocorticoid resistance syndromes -like ER22/23EK polymorphism-, while others, like bclI and N363S result in glucocorticoid hypersensitivity. In this study we hypothesized that single nucleotide gene polymorphism of the glucocorticoid receptor underlies different responsiveness to steroid therapy of patients diagnosed with Crohn`s disease and ulcerative colitis. Materials and Methods: we evaluated the incidence of BclI, N363S and ER 22/23EK polymorphisms of the glucocorticoid receptor in 103 patients of inflammatory bowel disease: 64 with Crohn`s disease(CD) and 39 with ulcerative colitis (UC). Mean age was 34.15 and 39.74 for CD and UC patients, respectively. Results: The incidence of polymorph BclI, N363S and ER22/23EK allels were 41.1%, 5.4% and 1.6% in Crohn disease and 29.5%, 6.4% and 2.6% in ulcerative colitis, respectively. Glucocorticoid responsive and glucocorticoid resistant groups were defined according to the patients responsiveness to glucocorticoid therapy. We found that in the glucocorticoid responsive IBD group the incidence of the ER22/23EK polymorphism that has decreased sensitivity to glucocorticoids was significantly lower (P=0.023) than in the healthy controls from the same geographical area. Conclusion: We found that the presence of the ER22/23EK polymorphism was significantly lower in glucocorticoid responsive patients with inflammatory bowel disease. M2082 Association Between Toll-Like 4, Cd14 Polymorphisms and Inflammatory Bowel Disease in Turkish Population Hakan Akin, Filiz T. Özdemir, Özlen Atug, Gulgun Tahan, Fatih Eren, Bilgi Baca, Ismail Hamzaoglu, Hulya O. Hamzaoglu, Nurdan Tozun Background: Inflammatory bowel disease (IBD) is a multifactorial disease with suspected background of genetic susceptibility. Increasing evidence suggest that innate immune system may have a key role in the pathogenesis of the disease. As a part of the innate immune system, Toll like receptors 4 (TLR4) and the related polymorphisms of A299G and T399I and the promotor of the CD 14 gene at position 159 are in candidate list accused for the pathogenesis of IBD. Aim: We want to investigate the relationship between the polymorphisms A299G, T399I and CD14 at position 159 with IBD. Methods: We collected clinical data and also blood samples for genetic analysis from the IBD pateints who are following in the IBD clinics of our hospital. The clinical data documented for the study was as follows; the type of the disease (Crohn's disease (CD) or Ulcerative Colitis(UC)), age, sex, date of the diagnosis, disease localization and phenotype, corticosteroid usage, smoking, family history, Enteamoba histolytica infestation history. For control group we include healty unrelated blood donors. Results: A total of 238 IBD patients (mean age: (41,7±15,5), Male/female: (127/111), 108 CD and 120 UC), and 191 controls ( age 35,2 ±11,2, Male/female:100/91 )included to the study. For all IBD patients the percent of presence of A299G, T399I heterozygot genotypes and for CD 14 (TT) genotype were 5.5% (13/238), 6,7%(16/238), 29,8% (71/238) respectively. These frequencies were not different statistically in the control group which were 5.9%, 7,5% and 30,9% respectively. Beside genotype frequencies, also the calculated allele frequencies were not different in two groups. In subgroup analysis we found that for patients with Crohn's disease, A299G heterozygote genotype more was frequent in the patients with ileal disease (p: 0,021) and also inflammatory genotype (p:0.008). Discussion: Our results related to the genotype and allele frequencies of TLR 4 polymorphisms in IBD is the first result for our country. M2083 Prediction of Crohn's Disease Aggression Through NOD2CARD15 Sequence Variation Maneesha Bhullar, Ken Sharpe, Paula Lewis, Bernadette Viney, Gregor Brown, Finlay A. Macrae, Margaret J. Smith Mutations in the NOD2/CARD15 gene predispose to Crohn's Disease (CD). This project aimed to investigate the association between NOD2/CARD15 sequence variants and disease progression in CD patients who have had ileo-caecal or small bowel resections. Method: Thirty unrelated CD patients (20 males; range 25-69 years) with initial caecal and/or small bowel resection during 1980-2000 were divided into two groups defined by a requirement A-465 AGA Abstracts for further surgery (Group 1; n=15) or not (Group 2; n=15). Clinical and demographic data were documented to December 2005, and all NOD2/CARD15 exons sequenced. Results: 16 sequence variations (9 with allele frequency >10%) have been identified. The allele frequencies for the pathogenic mutations R702W (Exon 4), G908R (Exon 8) and 3020insC (Exon 11) were 16.7 %, 3 % and 13.3 %; there were 12 patients with the P268S sequence variant, including all 8 patients with the pathogenic mutations. Patients with any pathogenic mutation had a significantly reduced time period between diagnosis and initial surgery (medians: 3020insC mutation 0.5 months, R702W or G908R 12.0 months and no mutation 48.5 months, p<0.027 Figure). There was also a trend to more operations in mutation positive patients (1.09 operations/100 months compared to 0.78 operations/100 months, p=0.096). Bi-allelic carriers did not have more aggressive disease than mono-allelic carriers, nor did patients with any of the unclassified variants in isolation or compound. Conclusion: This data suggests that the presence of a NOD2/CARD15 pathogenic mutation (especially 3020insC) predicts a more aggressive progression of disease. The P268S change occurred in linkage with pathogenic changes, without independent effect. If these results on post- operative relapse can be verified, early “top down” treatment in NOD2/CARD15 mutation carriers should be considered. M2084 Diagnostic Role of Biliary Pancreatic Elastase for Cholangiocarcinoma in Patients with Cholestasis Chiung-Yu Chen, Shu-Chu Shiesh, Wei-Lun Tsai Background and Aim: A wide array of proteins is secreted into bile and may be associated with biliary tract diseases. This study aimed to discover novel biomarker in bile for cholangi- ocarcinoma. Methods: Bile was collected from patients with bile duct obstruction. Proteins were separated by two-dimensional electrophoresis and identified by mass spectrometry. Levels of mRNA and protein expression of the candidate biomarker were analyzed by real- time PCR and Western blotting, respectively, whereas enzyme activity was measured by a kinetic method. The diagnostic efficacy was assessed by receiver operating characteristic (ROC) curve analysis. Results: Pancreatic elastase (PE) 3B was identified as a biomarker for cholangiocarcinoma. The mRNA of PE 3B was up-regulated in cancerous tissues, compared to non-cancerous tissues. The protein expression and enzyme activity of PE in bile were elevated in patients with cholangiocarcinoma, compared to gallstone patients. Biliary amylase activity was used to correct the presence of pancreaticobiliary reflux. Significantly elevated PE/amylase ratios in bile were found in patients with cholangiocarcinoma (0.214 ± 0.045) than those with gallstone (0.023 ± 0.005, p < 0.001). The area under the ROC curve of the ratio was 0.877 (95% CI: 0.765 to 0.988). Using 0.065 as a cutoff value, the ratio distinguished malignant from benign causes of biliary obstruction with a sensitivity of 82% and a specificity of 89%. Conclusion: Our present data demonstrate that PE in bile is a biomarker for cholangiocarcinoma and the combination measurement of PE and amylase enhances diagnostic efficacy. Receiver operating characteristic (ROC) curve of biliary pancreatic elastase, amylase and the pancreatic elastase/amylase ratio for the diagnosis of malignant biliary obstruction. M2085 Overexpression of the Gamma 2 Chain of Laminin-5 and Matrilysin in the Progression of Human Biliary Tract Cancers Hiroyuki Yamamoto, Shigeru Sasaki, Chie Miyamoto, Nobuki Miyamoto, Katsuhiko Nosho, Hiroaki Taniguchi, Tadateru Maehata, Kentaro Yamashita, Yasushi Adachi, Yoshiaki Arimura, Fumio Itoh, Kohzoh Imai, Yasuhisa Shinomura Despite advances in diagnosis and treatment, the prognosis of patients with biliary tract cancer is still poor. Biliary tract cancer is often unresectable at the time of diagnosis because of metastases to lymph nodes and distant organs, invasion to the liver, or peritoneal dissemina- tion. Identification of molecules involved in this cancer progression will convey therapeutic AGA Abstracts
Transcript of M2082 Association Between Toll-Like 4, Cd14 Polymorphisms and Inflammatory Bowel Disease in Turkish...
14% and 34%, respectively. Our aim in this study was to determine the allele frequenciesof the two disease-causing polymorphisms in a Swedish cohort of patients with CD. Methods:Blood samples were collected from 125 consecutive CD patients (64 men, 61 women). Thesamples were frozen and stored at -70 centigrade. DNA was isolated and amplified usingspecific primers and the Q705K polymorphism of CIAS1 and C10X polymorphism of CARD8were detected by the commercially available Megabace SNuPe genotyping kit. Moreover,the three most common polymorphisms of NOD2 were assessed, i.e. insC, 702, and 908,respectively. The results were compared to blood DNA bank controls (n=806). Results: Atleast one allele variant in the NOD2 gene was found in 16 % of CD patients, OR=2.32 (95% CI: 1.39-3.88). Ten percent of the patients carried heterozygote polymorphisms in bothof the CIAS1 and CARD8 genes, OR=2.34 (95 % CI: 1.13-4.84). None of these patientshad polymorphisms in the NOD2 gene. Conclusions: Compound polymorphisms of CIAS1and CARD8 doubled the susceptibility for CD in our cohort. This may be an importantgenetic risk factor in individuals with normal NOD2. However, the data needs confirmationin larger patient groups.
M2081
Glucocorticoid Receptor Polymorphism in Inflammatory Bowel DiseaseEmese Mihály, Péter Gergics, Pál Miheller, Károly Rácz, Zsolt Tulassay
Background and aims: Glucocorticoids are used to treat a wide variety of inflammatorydiseases. They are potent inhibitors of T cell activation and proinflammatory cytokines andeffectively used to treat active inflammatory bowel disease (IBD). However, there is largevariability in responsiveness and sensitivity between individuals to glucocorticoid therapy,both with regard to efficacy and the prevalence and severity of the side effects. Singlenucleotide gene polymorphism of the glucocorticoid receptor results in diverse clinicalmanifestations. Most of the mutations were found associated with glucocorticoid resistancesyndromes -like ER22/23EK polymorphism-, while others, like bclI and N363S result inglucocorticoid hypersensitivity. In this study we hypothesized that single nucleotide genepolymorphism of the glucocorticoid receptor underlies different responsiveness to steroidtherapy of patients diagnosed with Crohn`s disease and ulcerative colitis. Materials andMethods: we evaluated the incidence of BclI, N363S and ER 22/23EK polymorphisms ofthe glucocorticoid receptor in 103 patients of inflammatory bowel disease: 64 with Crohn`sdisease(CD) and 39 with ulcerative colitis (UC). Mean age was 34.15 and 39.74 for CD andUC patients, respectively. Results: The incidence of polymorph BclI, N363S and ER22/23EKallels were 41.1%, 5.4% and 1.6% in Crohn disease and 29.5%, 6.4% and 2.6% in ulcerativecolitis, respectively. Glucocorticoid responsive and glucocorticoid resistant groups weredefined according to the patients responsiveness to glucocorticoid therapy. We found thatin the glucocorticoid responsive IBD group the incidence of the ER22/23EK polymorphismthat has decreased sensitivity to glucocorticoids was significantly lower (P=0.023) than inthe healthy controls from the same geographical area. Conclusion: We found that the presenceof the ER22/23EK polymorphism was significantly lower in glucocorticoid responsive patientswith inflammatory bowel disease.
M2082
Association Between Toll-Like 4, Cd14 Polymorphisms and InflammatoryBowel Disease in Turkish PopulationHakan Akin, Filiz T. Özdemir, Özlen Atug, Gulgun Tahan, Fatih Eren, Bilgi Baca, IsmailHamzaoglu, Hulya O. Hamzaoglu, Nurdan Tozun
Background: Inflammatory bowel disease (IBD) is a multifactorial disease with suspectedbackground of genetic susceptibility. Increasing evidence suggest that innate immune systemmay have a key role in the pathogenesis of the disease. As a part of the innate immunesystem, Toll like receptors 4 (TLR4) and the related polymorphisms of A299G and T399Iand the promotor of the CD 14 gene at position 159 are in candidate list accused for thepathogenesis of IBD. Aim: We want to investigate the relationship between the polymorphismsA299G, T399I and CD14 at position 159 with IBD. Methods: We collected clinical dataand also blood samples for genetic analysis from the IBD pateints who are following in theIBD clinics of our hospital. The clinical data documented for the study was as follows; thetype of the disease (Crohn's disease (CD) or Ulcerative Colitis(UC)), age, sex, date of thediagnosis, disease localization and phenotype, corticosteroid usage, smoking, family history,Enteamoba histolytica infestation history. For control group we include healty unrelatedblood donors. Results: A total of 238 IBD patients (mean age: (41,7±15,5), Male/female:(127/111), 108 CD and 120 UC), and 191 controls ( age 35,2 ±11,2, Male/female:100/91)included to the study. For all IBD patients the percent of presence of A299G, T399Iheterozygot genotypes and for CD 14 (TT) genotype were 5.5% (13/238), 6,7%(16/238),29,8% (71/238) respectively. These frequencies were not different statistically in the controlgroup which were 5.9%, 7,5% and 30,9% respectively. Beside genotype frequencies, alsothe calculated allele frequencies were not different in two groups. In subgroup analysis wefound that for patients with Crohn's disease, A299G heterozygote genotype more was frequentin the patients with ileal disease (p: 0,021) and also inflammatory genotype (p:0.008).Discussion: Our results related to the genotype and allele frequencies of TLR 4 polymorphismsin IBD is the first result for our country.
M2083
Prediction of Crohn's Disease Aggression Through NOD2CARD15 SequenceVariationManeesha Bhullar, Ken Sharpe, Paula Lewis, Bernadette Viney, Gregor Brown, Finlay A.Macrae, Margaret J. Smith
Mutations in the NOD2/CARD15 gene predispose to Crohn's Disease (CD). This projectaimed to investigate the association between NOD2/CARD15 sequence variants and diseaseprogression in CD patients who have had ileo-caecal or small bowel resections. Method:Thirty unrelated CD patients (20 males; range 25-69 years) with initial caecal and/or smallbowel resection during 1980-2000 were divided into two groups defined by a requirement
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A-465 AGA Abstracts
for further surgery (Group 1; n=15) or not (Group 2; n=15). Clinical and demographic datawere documented to December 2005, and all NOD2/CARD15 exons sequenced. Results:16 sequence variations (9 with allele frequency >10%) have been identified. The allelefrequencies for the pathogenic mutations R702W (Exon 4), G908R (Exon 8) and 3020insC(Exon 11) were 16.7 %, 3 % and 13.3 %; there were 12 patients with the P268S sequencevariant, including all 8 patients with the pathogenic mutations. Patients with any pathogenicmutation had a significantly reduced time period between diagnosis and initial surgery(medians: 3020insC mutation 0.5 months, R702W or G908R 12.0 months and no mutation48.5 months, p<0.027 Figure). There was also a trend to more operations in mutationpositive patients (1.09 operations/100 months compared to 0.78 operations/100 months,p=0.096). Bi-allelic carriers did not have more aggressive disease than mono-allelic carriers,nor did patients with any of the unclassified variants in isolation or compound. Conclusion:This data suggests that the presence of a NOD2/CARD15 pathogenic mutation (especially3020insC) predicts a more aggressive progression of disease. The P268S change occurredin linkage with pathogenic changes, without independent effect. If these results on post-operative relapse can be verified, early “top down” treatment in NOD2/CARD15 mutationcarriers should be considered.
M2084
Diagnostic Role of Biliary Pancreatic Elastase for Cholangiocarcinoma inPatients with CholestasisChiung-Yu Chen, Shu-Chu Shiesh, Wei-Lun Tsai
Background and Aim: A wide array of proteins is secreted into bile and may be associatedwith biliary tract diseases. This study aimed to discover novel biomarker in bile for cholangi-ocarcinoma. Methods: Bile was collected from patients with bile duct obstruction. Proteinswere separated by two-dimensional electrophoresis and identified by mass spectrometry.Levels of mRNA and protein expression of the candidate biomarker were analyzed by real-time PCR and Western blotting, respectively, whereas enzyme activity was measured by akinetic method. The diagnostic efficacy was assessed by receiver operating characteristic(ROC) curve analysis. Results: Pancreatic elastase (PE) 3B was identified as a biomarker forcholangiocarcinoma. The mRNA of PE 3B was up-regulated in cancerous tissues, comparedto non-cancerous tissues. The protein expression and enzyme activity of PE in bile wereelevated in patients with cholangiocarcinoma, compared to gallstone patients. Biliary amylaseactivity was used to correct the presence of pancreaticobiliary reflux. Significantly elevatedPE/amylase ratios in bile were found in patients with cholangiocarcinoma (0.214 ± 0.045)than those with gallstone (0.023 ± 0.005, p < 0.001). The area under the ROC curve ofthe ratio was 0.877 (95% CI: 0.765 to 0.988). Using 0.065 as a cutoff value, the ratiodistinguished malignant from benign causes of biliary obstruction with a sensitivity of 82%and a specificity of 89%. Conclusion: Our present data demonstrate that PE in bile is abiomarker for cholangiocarcinoma and the combination measurement of PE and amylaseenhances diagnostic efficacy.
Receiver operating characteristic (ROC) curve of biliary pancreatic elastase, amylase and thepancreatic elastase/amylase ratio for the diagnosis of malignant biliary obstruction.
M2085
Overexpression of the Gamma 2 Chain of Laminin-5 and Matrilysin in theProgression of Human Biliary Tract CancersHiroyuki Yamamoto, Shigeru Sasaki, Chie Miyamoto, Nobuki Miyamoto, KatsuhikoNosho, Hiroaki Taniguchi, Tadateru Maehata, Kentaro Yamashita, Yasushi Adachi,Yoshiaki Arimura, Fumio Itoh, Kohzoh Imai, Yasuhisa Shinomura
Despite advances in diagnosis and treatment, the prognosis of patients with biliary tractcancer is still poor. Biliary tract cancer is often unresectable at the time of diagnosis becauseof metastases to lymph nodes and distant organs, invasion to the liver, or peritoneal dissemina-tion. Identification of molecules involved in this cancer progression will convey therapeutic
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