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Progenitor Cells
AGA Abstracts
between groups. Parenchymal and ductal abnormalities were similar except for a higher prevalence of pancreas divisum in MUT+ (25.6%) v. MUT- (12.6%) individuals (p=0.06). MUT+ individuals were more likely to have a history of acute recurrent pancreatitis (66.7%) compared to MUT- (43.2%, p=0.01), but chronic pancreatitis was similar (MUT+, 26.3%; MUT- 36.9%, p=0.23). Family history of pancreatic cancer was similar in both groups (p= 0.59), but MUT+ individuals were more likely to have a family history of acute pancreatitis (p=0.003) and chronic pancreatitis (p=0.05). Variables meeting criteria for the regression model and associated with having a positive genetic mutation included pancreas divisum (OR 2.81, p,0.04), family history of acute pancreatitis (3.46, p=0.01) and personal history of acute recurrent pancreatitis (2.82, p=0.01). Conclusion: In adults with known or suspected pancreatic disease who undergo genetic testing, factors associated with a positive mutation include pancreas divisum, a personal history of acute recurrent pancreatitis, and family history of acute pancreatitis. Patient characteristics
Su1332 CFTR, PRSS1, and SPINK1 Variants in Japanese Patients With Pancreatitis Hiroyuki Ariga, Kiyoshi Kume, Atsushi Masamune, Tooru Shimosegawa Background: Cystic fibrosis (CF) is an autosomal recessive disease caused by variants in the CFTR gene. To date, over 1,700 variants in CFTR have been identified. The incidence of CF in Japanese is very low (1 per 350,000 live births) compared with Caucasian (1 per 3,000 live births). The most common variants of CFTR gene in Caucasian, such as F508del, are rare in Japanese. The profiles of CFTR variants in Japanese are different from those of Caucasian. Several genes, such as cationic trypsinogen (PRSS1) and serine protease inhibitor Kazal type 1 (SPINK1), play an important role in pancreatitis susceptibility, but the roles of CFTR variants in pancreatitis have been unclear in Japan. IVS8-5T, p.Q1352H, and p.R1453W in the CFTR gene were reported to be found frequently in Asian population and affected the function of the CFTR protein. Previous studies in the Japanese population reported an association between these variants and pancreatitis, but they were analyzed at a small scale. Methods: This study was approved by the ethics committee of Tohoku University Graduate School of Medicine. A total of 482 patients (271 chronic pancreatitis (CP), 211 acute pancreatitis (AP)) and 440 controls were enrolled in this study. p.Q1352H and p.R1453W in the CFTR gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). IVS8-5T in the CFTR gene was analyzed by SNP genotyping with Real-Time PCR and direct sequencing. PRSS1 and SPINK1 variants were analyzed by PCR-RFLP and direct sequencing. Results: p.Q1352H in the CFTR gene was found in 17 (6.3%) of 271 patients with CP, 13 (6.2%) of 211 with AP, and 19 (4.3%) of 440 controls. p.R1453W in the CFTR gene was found in 16 (5.9%) of 271 with CP, 6 (2.8%) of 211 with AP, and 16 (3.6%) of 440 controls. IVS8-5T in the CFTR gene was found in 10 (3.7%) of 271 with CP, 11 (5.2%) of 211 with AP, and 15 (3.9%) of 385 controls. The CFTR variants were not significantly associated with CP (p=0.20, odds ratio (OR) 1.4, 95% confidence interval (CI) 0.9-2.1) or with AP (p=0.52, OR 1.2, 95%CI 0.71.9). p.R122H and p.N29I in the PRSS1 gene were found in 8 (4.7%) and 1 (0.6%) of 170 with CP, respectively. PRSS1 variants were not found in 440 controls. The PRSS1 variants were significantly associated with CP (p ,0.01, OR 49.2, 95%CI 4.9-494.1). p.N34S and c.194+2T.C in the SPINK1 gene were found in 19 (7.0%) and 17 (6.3%) of 271 with CP, respectively. SPINK1 variants were found in 2 (0.05%) of 440 controls. The SPINK1 variants were significantly associated with CP (p ,0.001, OR 30.4, 95%CI 8.0-115.4). Conclusions: PRSS1 and SPINK1 had a strong association with CP. But, the accumulation of CFTR variants in pancreatitis was less pronounced than previous studies in Japan.
† All numbers are percentiles with 95% confidence intervals unless otherwise specified. Su1723 Helicobacter Infection Promotes Mouse Gastric Organoid Growth, and Altered the Property of Gastric Stem/Progenitor Cells Wataru Shibata, Shin Maeda Helicobacter-related chronic gastritis is the major predisposing factor for gastric carcinogenesis. However, precise mechanism from chronic gastritis to gastric cancer is not fully elucidated yet. Recently, cancer stem cell concept has been proposed as an attractive theory to explain cancer development. Because of similar characteristics, cancer stem cells may be derived from normal tissue stem/progenitor cells. In order to analyze an effect of chronic gastric inflammation on gastric stem cells and to examine a relationship between the stem cells and tumorigenesis, we used the gastric organoid culture system which can be generated from single stem/progenitor cell of mouse gastric mucosa. Mice infected with Helicobacter felis were sacrificed at 6 months post infection. Histological analysis showed severe gastric inflammation with lymph follicles, neck cell hyperplasia, oxyntic atrophy, and mucous metaplasia especially in gastric corpus. Numbers of organoids from Helicobacter-infected gastric mucosa were significantly higher than those from uninfected gastric mucosa. In addition, organoid growth was more rapid in those isolated from Helicobacter-infected mucosa compared to the control. mRNA expression was assessed by microarray analysis and realtime PCR, and found that stem cell markers such as CD44 and DCAMKL1 and genes associated with intestinal metaplasia such as villin-1 or intestine specific homeobox were increased in organoids cultured from Helicobacter-infected mucosa compared to the control. These expressions were also confirmed by immunohitochemistry in tissue samples. These results suggested that Helicobacter infection increased the number of stem cells and altered the property of stem cells toward intestinal metaplasia. We further investigated the tumorigenisity of the organoids from Helicobacter-infected or chemical carcinogen methyl-nitroso-urea (MNU) treated mucosa by implanting them into immune deficient mice. After several months, tumor was generated from the organoids cultured from MNU-injected mucosa, whereas organoids from Helicobacter-infected mucosa did not give rise to tumors. The current study suggests that one of the important functions of Helicobacter-induced inflammation increases the gastric stem/progenitor cells with altering the property of gastric stem/progenitor cells resembling intestinal epithelial lineage, which may become an origin of cancer initiating cells. Organoid culture system and their combined xenograft model are invaluable tools for research of gastric carcinogenesis.
Su1333 Testing Adults With Pancreatic Disease for Genetic Abnormalities: Factors Associated With Having a Positive Genetic Test Darren D. Ballard, Joyce R. Flueckiger, Evan L. Fogel, Lee McHenry, Glen A. Lehman, James L. Watkins, Stuart Sherman, Gregory A. Cote Background: Genetic predisposition for acute recurrent and chronic pancreatitis includes mutations in CFTR, PRSS1, SPINK1 and CTRC genes. We sought to differentiate demographic and phenotypic features of individuals with adult-onset pancreatic disease with and without pancreas-specific genetic mutations. Methods: Retrospective cohort study of all individuals ≥ 18 years of age who underwent testing for CFTR, PRSS1, SPINK1 and CTRC gene mutations in an adult pancreatic disease clinic between 1/2010 and 8/2012. Patients were evaluated for genetic abnormalities in the setting of idiopathic acute recurrent or chronic pancreatitis, or for pancreatic-type symptoms in conjunction with a family history of pancreatic disease. Patients were classified as having a genetic mutation (MUT+) or not (MUT-). We abstracted medical records for patient demographics, family history of pancreatitis or pancreatic cancer, and presence of pain, exocrine and endocrine insufficiency. Finally, we compared morphologic characteristics including the presence of pancreas divisum, pancreatic calcifications, and pancreatic duct stricture. Variables having a potential association on univariate analysis (p,0.10) were incorporated into a forward stepwise regression model, dropping variables having a p ,0.10. Results: During the study period, 150 subjects underwent testing for pancreas-specific genetic mutations, 39 (26%) having one or more mutations identified (MUT+) in CFTR (n=25), SPINK1 (n=11), PRSS1 (n=9), CTRC (n=5), or some combination. Mean age of presentation (MUT-, 45.5±15.8; MUT+ 42.2±15.4, p=0.27) were similar but mean age of symptom onset being earlier among MUT+ (34.1±16.7) compared to MUT(39.8±16.5, p=0.07) (table). Other demographic features and symptoms/signs were similar
AGA Abstracts
Su1724 Expression of CD44 Variant 9 Contributes to 5-Fluorouracil Resistance in Gastric Cancer Through Anti-Oxidative Stress Mechanism Sawako Okada, Hidekazu Suzuki, Tatsuhiro Masaoka, Hitoshi Tsugawa, Juntaro Matsuzaki, Seiichiro Fukuhara, Kenro Hirata, Hideki Mori, Hideyuki Saya, Toshifumi Hibi Background. Cancer stem cell (CSC), which has capacities for self-renewal and differentiation, has recently identified. Large amount of data support that CSCs may be associated with chemoresistance, radioresistance and recurrence of cancers. CD44 is known as a specific cell surface marker of gastric cancer stem cells. CD44 variant 9 (CD44v9), one of variant isoforms of CD44, is reported that it has association with prognosis, clinical stage, and
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between groups. Parenchymal and ductal abnormalities were similar except for a higher prevalence of pancreas divisum in MUT+ (25.6%) v. MUT- (12.6%) individuals (p=0.06). MUT+ individuals were more likely to have a history of acute recurrent pancreatitis (66.7%) compared to MUT- (43.2%, p=0.01), but chronic pancreatitis was similar (MUT+, 26.3%; MUT- 36.9%, p=0.23). Family history of pancreatic cancer was similar in both groups (p= 0.59), but MUT+ individuals were more likely to have a family history of acute pancreatitis (p=0.003) and chronic pancreatitis (p=0.05). Variables meeting criteria for the regression model and associated with having a positive genetic mutation included pancreas divisum (OR 2.81, p,0.04), family history of acute pancreatitis (3.46, p=0.01) and personal history of acute recurrent pancreatitis (2.82, p=0.01). Conclusion: In adults with known or suspected pancreatic disease who undergo genetic testing, factors associated with a positive mutation include pancreas divisum, a personal history of acute recurrent pancreatitis, and family history of acute pancreatitis. Patient characteristics
Su1332 CFTR, PRSS1, and SPINK1 Variants in Japanese Patients With Pancreatitis Hiroyuki Ariga, Kiyoshi Kume, Atsushi Masamune, Tooru Shimosegawa Background: Cystic fibrosis (CF) is an autosomal recessive disease caused by variants in the CFTR gene. To date, over 1,700 variants in CFTR have been identified. The incidence of CF in Japanese is very low (1 per 350,000 live births) compared with Caucasian (1 per 3,000 live births). The most common variants of CFTR gene in Caucasian, such as F508del, are rare in Japanese. The profiles of CFTR variants in Japanese are different from those of Caucasian. Several genes, such as cationic trypsinogen (PRSS1) and serine protease inhibitor Kazal type 1 (SPINK1), play an important role in pancreatitis susceptibility, but the roles of CFTR variants in pancreatitis have been unclear in Japan. IVS8-5T, p.Q1352H, and p.R1453W in the CFTR gene were reported to be found frequently in Asian population and affected the function of the CFTR protein. Previous studies in the Japanese population reported an association between these variants and pancreatitis, but they were analyzed at a small scale. Methods: This study was approved by the ethics committee of Tohoku University Graduate School of Medicine. A total of 482 patients (271 chronic pancreatitis (CP), 211 acute pancreatitis (AP)) and 440 controls were enrolled in this study. p.Q1352H and p.R1453W in the CFTR gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). IVS8-5T in the CFTR gene was analyzed by SNP genotyping with Real-Time PCR and direct sequencing. PRSS1 and SPINK1 variants were analyzed by PCR-RFLP and direct sequencing. Results: p.Q1352H in the CFTR gene was found in 17 (6.3%) of 271 patients with CP, 13 (6.2%) of 211 with AP, and 19 (4.3%) of 440 controls. p.R1453W in the CFTR gene was found in 16 (5.9%) of 271 with CP, 6 (2.8%) of 211 with AP, and 16 (3.6%) of 440 controls. IVS8-5T in the CFTR gene was found in 10 (3.7%) of 271 with CP, 11 (5.2%) of 211 with AP, and 15 (3.9%) of 385 controls. The CFTR variants were not significantly associated with CP (p=0.20, odds ratio (OR) 1.4, 95% confidence interval (CI) 0.9-2.1) or with AP (p=0.52, OR 1.2, 95%CI 0.71.9). p.R122H and p.N29I in the PRSS1 gene were found in 8 (4.7%) and 1 (0.6%) of 170 with CP, respectively. PRSS1 variants were not found in 440 controls. The PRSS1 variants were significantly associated with CP (p ,0.01, OR 49.2, 95%CI 4.9-494.1). p.N34S and c.194+2T.C in the SPINK1 gene were found in 19 (7.0%) and 17 (6.3%) of 271 with CP, respectively. SPINK1 variants were found in 2 (0.05%) of 440 controls. The SPINK1 variants were significantly associated with CP (p ,0.001, OR 30.4, 95%CI 8.0-115.4). Conclusions: PRSS1 and SPINK1 had a strong association with CP. But, the accumulation of CFTR variants in pancreatitis was less pronounced than previous studies in Japan.
† All numbers are percentiles with 95% confidence intervals unless otherwise specified. Su1723 Helicobacter Infection Promotes Mouse Gastric Organoid Growth, and Altered the Property of Gastric Stem/Progenitor Cells Wataru Shibata, Shin Maeda Helicobacter-related chronic gastritis is the major predisposing factor for gastric carcinogenesis. However, precise mechanism from chronic gastritis to gastric cancer is not fully elucidated yet. Recently, cancer stem cell concept has been proposed as an attractive theory to explain cancer development. Because of similar characteristics, cancer stem cells may be derived from normal tissue stem/progenitor cells. In order to analyze an effect of chronic gastric inflammation on gastric stem cells and to examine a relationship between the stem cells and tumorigenesis, we used the gastric organoid culture system which can be generated from single stem/progenitor cell of mouse gastric mucosa. Mice infected with Helicobacter felis were sacrificed at 6 months post infection. Histological analysis showed severe gastric inflammation with lymph follicles, neck cell hyperplasia, oxyntic atrophy, and mucous metaplasia especially in gastric corpus. Numbers of organoids from Helicobacter-infected gastric mucosa were significantly higher than those from uninfected gastric mucosa. In addition, organoid growth was more rapid in those isolated from Helicobacter-infected mucosa compared to the control. mRNA expression was assessed by microarray analysis and realtime PCR, and found that stem cell markers such as CD44 and DCAMKL1 and genes associated with intestinal metaplasia such as villin-1 or intestine specific homeobox were increased in organoids cultured from Helicobacter-infected mucosa compared to the control. These expressions were also confirmed by immunohitochemistry in tissue samples. These results suggested that Helicobacter infection increased the number of stem cells and altered the property of stem cells toward intestinal metaplasia. We further investigated the tumorigenisity of the organoids from Helicobacter-infected or chemical carcinogen methyl-nitroso-urea (MNU) treated mucosa by implanting them into immune deficient mice. After several months, tumor was generated from the organoids cultured from MNU-injected mucosa, whereas organoids from Helicobacter-infected mucosa did not give rise to tumors. The current study suggests that one of the important functions of Helicobacter-induced inflammation increases the gastric stem/progenitor cells with altering the property of gastric stem/progenitor cells resembling intestinal epithelial lineage, which may become an origin of cancer initiating cells. Organoid culture system and their combined xenograft model are invaluable tools for research of gastric carcinogenesis.
Su1333 Testing Adults With Pancreatic Disease for Genetic Abnormalities: Factors Associated With Having a Positive Genetic Test Darren D. Ballard, Joyce R. Flueckiger, Evan L. Fogel, Lee McHenry, Glen A. Lehman, James L. Watkins, Stuart Sherman, Gregory A. Cote Background: Genetic predisposition for acute recurrent and chronic pancreatitis includes mutations in CFTR, PRSS1, SPINK1 and CTRC genes. We sought to differentiate demographic and phenotypic features of individuals with adult-onset pancreatic disease with and without pancreas-specific genetic mutations. Methods: Retrospective cohort study of all individuals ≥ 18 years of age who underwent testing for CFTR, PRSS1, SPINK1 and CTRC gene mutations in an adult pancreatic disease clinic between 1/2010 and 8/2012. Patients were evaluated for genetic abnormalities in the setting of idiopathic acute recurrent or chronic pancreatitis, or for pancreatic-type symptoms in conjunction with a family history of pancreatic disease. Patients were classified as having a genetic mutation (MUT+) or not (MUT-). We abstracted medical records for patient demographics, family history of pancreatitis or pancreatic cancer, and presence of pain, exocrine and endocrine insufficiency. Finally, we compared morphologic characteristics including the presence of pancreas divisum, pancreatic calcifications, and pancreatic duct stricture. Variables having a potential association on univariate analysis (p,0.10) were incorporated into a forward stepwise regression model, dropping variables having a p ,0.10. Results: During the study period, 150 subjects underwent testing for pancreas-specific genetic mutations, 39 (26%) having one or more mutations identified (MUT+) in CFTR (n=25), SPINK1 (n=11), PRSS1 (n=9), CTRC (n=5), or some combination. Mean age of presentation (MUT-, 45.5±15.8; MUT+ 42.2±15.4, p=0.27) were similar but mean age of symptom onset being earlier among MUT+ (34.1±16.7) compared to MUT(39.8±16.5, p=0.07) (table). Other demographic features and symptoms/signs were similar
AGA Abstracts
Su1724 Expression of CD44 Variant 9 Contributes to 5-Fluorouracil Resistance in Gastric Cancer Through Anti-Oxidative Stress Mechanism Sawako Okada, Hidekazu Suzuki, Tatsuhiro Masaoka, Hitoshi Tsugawa, Juntaro Matsuzaki, Seiichiro Fukuhara, Kenro Hirata, Hideki Mori, Hideyuki Saya, Toshifumi Hibi Background. Cancer stem cell (CSC), which has capacities for self-renewal and differentiation, has recently identified. Large amount of data support that CSCs may be associated with chemoresistance, radioresistance and recurrence of cancers. CD44 is known as a specific cell surface marker of gastric cancer stem cells. CD44 variant 9 (CD44v9), one of variant isoforms of CD44, is reported that it has association with prognosis, clinical stage, and
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