- Email: [email protected]
Applying Proteomics Integrated With Gene Microarray for Discovery of Colorectal Cancer Novel Biomarkers
January 2017
Abstracts
1
JC School of Public Health and Primary Care, Faculty of Medicine, Chinese University of Hong Kong, Hong Kong SAR, China, 2Ruijin Hospital North, Shanghai Jiaotong University, Shanghai, China, and 3School of Public Health, Sun Yat-sen University, Guangzhou, Guangdong, China BACKGROUND: To tailor and enhance rational utilization of
screening colonoscopy, prediction models for advanced proximal neoplasia (APN) based on age, gender and distal findings at sigmoidoscopy has been proposed.1 However, its discriminatory capability in Chinese populations remain unexplored. We aim to develop and validate a new model including distal findings to predict APN in a Chinese asymptomatic population. METHODS: Age, gender, and colonoscopic findings were collected prospectively (2013-2015) in a hospital-based endoscopy unit in Shanghai, China. We randomly divided eligible cases into a derivation (n¼3,839) and validation group (n¼1,994). A new index for risk-stratification of APN and its cut-off value were generated from the derivation group by binary logistic regression analyses. The area under the curve (AUC) was used to examine the predictive capability of the new index in the validation group. RESULTS: Age, gender and distal finding were all independent predictors of APN in both groups (all p value <0.01). The AUC of both groups and the adjusted odds ratios in the derivation groups are listed in Table 1. The low-risk group in the derivation group had an APN rate of 0.7%. In the validation group, 93.5% of APNs could be detected by using colonoscopy to screen intermediate and high risk groups. In total, using this index to select endoscopy could save 40.7% of colonoscopy procedures. Among subjects in the high risk group, 1 APN could be detected for every 13 colonoscopy procedures performed. CONCLUSION: The new index is a useful model for riskstratification of APN in Chinese population. (250 words) Conflicts of interest: The authors disclose no conflicts.
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PNPLA3 Gene Polymorphisms and the Associated Risk of Non-Alcoholic Fatty Liver Disease in Indian Subjects Rekha Singhal,1 and Manvi Vernekar,1 Deepak Amarapurkar2 1 Food Engineering and Technology Department, Institute of Chemical Technology, India, and 2Department of Gastroenterology, Bombay Hospital and Medical Research Centre, India
BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is
considered to be the hepatic manifestation of the Metabolic Syndrome. However, genetic predisposition of NAFLD has not been extensively studied in the Asian Indian population. METHODS: A retrospective case-control study with subjects diagnosed with fatty liver on ultrasound evaluation were recruited as cases (n¼100) and subjects with normal liver ultrasound were recruited as controls (n¼105). Anthropometric measures like age and BMI were recorded while biochemical parameters like lipid and liver profiling were carried out. Polymerase chain reaction using specific primers with subsequent restriction fragment length polymorphism technique was employed for genotyping. Genotype frequencies were evaluated by chisquare test. Student’s t-test was applied for comparison of continuous variables. Odds ratios (ORs) were adjusted for age, gender, and BMI by logistic regression analysis. RESULTS: A total of 205 subjects were recruited with 105 controls (mean age 39.3512.07 years) and 100 cases (mean age 41.310.40 years). BMI of cases (26.574.53) was significantly higher than that of controls (23.763.57) (p <0.0001). AST and ALT levels were also significantly elevated in cases (p<0.05). Rs738409 was found to be in genotypic and allelic (p<0.05) association with NAFLD. The frequency of minor allele G in cases was 0.40 which was significantly greater than that of controls at 0.25 indicating a very likely association with NAFLD (additive model, p<0.05). The odds ratio (95% confidence interval) adjusted for age, gender, and BMI was 3.271 (1.22-8.75). CONCLUSION: Corroborating with the data, high BMI and elevated AST and ALT levels may increase susceptibility to NAFLD. Also, the G/G genotype and G allele of rs738409 was found to be significantly associated with NAFLD in Indian subjects. Conflicts of interest: The authors disclose no conflicts.
Applying Proteomics Integrated With Gene Microarray for Discovery of Colorectal Cancer Novel Biomarkers Xiao-fen Li,1 Jie-kai Yu,1 Wang-xiong Hu,1 and Shu Zheng1
Table 1. Components of a new index to predict advanced proximal neoplasia 1
1. Imperiale TF, Wagner DR, Lin CY, Larkin GN, Rogge JD, Ransohoff DF. Using Risk for Advanced Proximal Colonic Neoplasia to Tailor Endoscopic Screening for Colorectal Cancer. Ann Intern Med 2003; 139: 959-96510
Cancer Institute, the Second Affiliated Hospital, Zhejiang University School of Medicine, China BACKGROUND: Colorectal cancer (CRC) is the fourth leading
cause of cancer-related deaths worldwide. Proteins, as executives of genes’ instructions, are responsible for
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Abstracts
Clinical Gastroenterology and Hepatology Vol. 15, No. 1
cellular phenotypes. Integrating proteomics with gene microarray, we conducted this study to identify potential protein biomarkers of CRC. METHODS: Isobaric tags with related and absolute quantitation (iTRAQ) labeling mass spectrometry (MS) was applied to screen and identify differentially expressed proteins between paired CRC and adjacent normal mucosa. Meanwhile, Affymetrix U133plus2.0 microarrays were used to perform gene microarray analysis. Verification experiments included immunohistochemistry (IHC) and western blot of selected proteins. RESULTS: Overall, 5469 differentially expressed proteins were detected with iTRAQ-MS from 24 matched CRC and adjacent normal tissues. And gene microarray identified 39859 differential genes from 52 patients. Of these, 3083 differential proteins had corresponding differentially expressed genes, with 245 proteins and their genes showed >1.5-fold change in expression level. Gene ontology enrichment analysis revealed that up-regulated proteins were more involved in cell adhesion and motion than down-regulated proteins. In addition, up-regulated proteins were more likely to be located in nucleus and vesicles. Further verification experiments with IHC and western blot confirmed differential expression levels of 5 proteins (S100 calcium-binding protein A9, annexin A3, nicotinamide phosphoribosyltransferase, carboxylesterase 2 and calcium activated chloride channel A1) between CRC and normal tissues. CONCLUSION: S100 calcium-binding protein A9, annexin A3, nicotinamide phosphoribosyltransferase, carboxylesterase 2 and calcium activated chloride channel A1 were probably potential biomarkers of colorectal cancer. Annexin A3 may be associated with colorectal carcinogenesis.
of gastroduodenal mucosal injury would be assessed according to the modified Lanza Endoscopic Scoring System. RESULTS: From August 2015 to February 2016, 109 of 124 patients finished the follow-up, of whom 92.7% (101/ 109) underwent EGD. The prevalence of multiple gastroduodenal lesions, which should be treated with PPIs, is higher in ticagrelor plus aspirin group (38.5%, 15/39), compared with that in aspirin (33.3%, 10/30) or ticagrelor (31.3%, 10/32) monotherapy group, respectively. All patients were prescribed with PPIs for 1 month, 71.2% (72/101) of whom were treated with 3-month PPIs, only 7.0% (7/101) with more than 6-month. Among 17 patients with history of peptic ulcers, 17.6% (3/17) were treated with 1-month PPIs, 64.7% (11/17) with 3month, only 17.7% (3/17) with more than 6-month. CONCLUSION: It seems that Chinese patients are rarely under concomitant use of PPIs with antiplatelet therapy for more than 6 months, even those with history of peptic ulcer, demonstrating an unmet need for effective prophylactic treatment with PPIs to decrease the risk of upper GI bleeding.
Conflicts of interest: The authors disclose no conflicts.
Conflicts of interest: The authors disclose no conflicts.
Current Status of Concomitant Use of Proton Pump Inhibitors With Different Antiplatelet Therapies in Chinese Patients After Coronary Artery Bypass Grafting Chenyue Tang,1 Yunpeng Zhu,2 Cong Ye,2 Qiang Zhao,2 and Lifen Yu1 1
2
Department of Gastroenterology, and Cardiac Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, China BACKGROUND: Gastrointestinal complications of antiplate-
let drugs are common. Investigating the current status of concomitant use of proton pump inhibitors (PPIs) is needed in antiplatelet drug users after coronary artery bypass grafting (CABG). METHODS: Two hundred and seventy patients enrolled in Ruijin hospital, maximum proportion of those who were randomly assigned into three groups after CABG including aspirin or ticagrelor monotherapy group and ticagrelor plus aspirin group (DACAB trial, NCT 02201771), would undergo esophagogastroduodenoscopy (EGD) with their consent at 12th month. The grade
Capsule Endoscopy in Intestinal Lymphagiectasia: A Novel Endoscopic Classification and its Clinical Correlation Kui-liang Liu,1 Fang-yun Liu,1 Wen-bin Shen,2 and Jing Wu1 1
Department of Gastroenterology, Beijing Shijitan Hospital, Capital Medical University, and 2Department of Lymphatic Surgery, Beijing Shijitan Hospital, Capital Medical University BACKGROUND: Capsule Endoscopy (CE) is useful in diag-
nosis of intestinal lymphagiectasia (IL). However, systemic analysis of endoscopic and clinical characteristics remains unavailable. METHODS: Review the endoscopic characteristics of patients underwent CE in our endoscopy center between 2011 and 2015 and develop a novel endoscopic classification to investigate its clinical correlation. RESULTS: A total of 54 patients, 29 males and 25 females with median age of 24 (4-56) and duration lasting 20 days to 23 years were included. 42 cases were diagnosed as primary IL. Typical endoscopic characteristics included scattering, grouping or diffuse white-yellow
Abstracts
1
JC School of Public Health and Primary Care, Faculty of Medicine, Chinese University of Hong Kong, Hong Kong SAR, China, 2Ruijin Hospital North, Shanghai Jiaotong University, Shanghai, China, and 3School of Public Health, Sun Yat-sen University, Guangzhou, Guangdong, China BACKGROUND: To tailor and enhance rational utilization of
screening colonoscopy, prediction models for advanced proximal neoplasia (APN) based on age, gender and distal findings at sigmoidoscopy has been proposed.1 However, its discriminatory capability in Chinese populations remain unexplored. We aim to develop and validate a new model including distal findings to predict APN in a Chinese asymptomatic population. METHODS: Age, gender, and colonoscopic findings were collected prospectively (2013-2015) in a hospital-based endoscopy unit in Shanghai, China. We randomly divided eligible cases into a derivation (n¼3,839) and validation group (n¼1,994). A new index for risk-stratification of APN and its cut-off value were generated from the derivation group by binary logistic regression analyses. The area under the curve (AUC) was used to examine the predictive capability of the new index in the validation group. RESULTS: Age, gender and distal finding were all independent predictors of APN in both groups (all p value <0.01). The AUC of both groups and the adjusted odds ratios in the derivation groups are listed in Table 1. The low-risk group in the derivation group had an APN rate of 0.7%. In the validation group, 93.5% of APNs could be detected by using colonoscopy to screen intermediate and high risk groups. In total, using this index to select endoscopy could save 40.7% of colonoscopy procedures. Among subjects in the high risk group, 1 APN could be detected for every 13 colonoscopy procedures performed. CONCLUSION: The new index is a useful model for riskstratification of APN in Chinese population. (250 words) Conflicts of interest: The authors disclose no conflicts.
e39
PNPLA3 Gene Polymorphisms and the Associated Risk of Non-Alcoholic Fatty Liver Disease in Indian Subjects Rekha Singhal,1 and Manvi Vernekar,1 Deepak Amarapurkar2 1 Food Engineering and Technology Department, Institute of Chemical Technology, India, and 2Department of Gastroenterology, Bombay Hospital and Medical Research Centre, India
BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is
considered to be the hepatic manifestation of the Metabolic Syndrome. However, genetic predisposition of NAFLD has not been extensively studied in the Asian Indian population. METHODS: A retrospective case-control study with subjects diagnosed with fatty liver on ultrasound evaluation were recruited as cases (n¼100) and subjects with normal liver ultrasound were recruited as controls (n¼105). Anthropometric measures like age and BMI were recorded while biochemical parameters like lipid and liver profiling were carried out. Polymerase chain reaction using specific primers with subsequent restriction fragment length polymorphism technique was employed for genotyping. Genotype frequencies were evaluated by chisquare test. Student’s t-test was applied for comparison of continuous variables. Odds ratios (ORs) were adjusted for age, gender, and BMI by logistic regression analysis. RESULTS: A total of 205 subjects were recruited with 105 controls (mean age 39.3512.07 years) and 100 cases (mean age 41.310.40 years). BMI of cases (26.574.53) was significantly higher than that of controls (23.763.57) (p <0.0001). AST and ALT levels were also significantly elevated in cases (p<0.05). Rs738409 was found to be in genotypic and allelic (p<0.05) association with NAFLD. The frequency of minor allele G in cases was 0.40 which was significantly greater than that of controls at 0.25 indicating a very likely association with NAFLD (additive model, p<0.05). The odds ratio (95% confidence interval) adjusted for age, gender, and BMI was 3.271 (1.22-8.75). CONCLUSION: Corroborating with the data, high BMI and elevated AST and ALT levels may increase susceptibility to NAFLD. Also, the G/G genotype and G allele of rs738409 was found to be significantly associated with NAFLD in Indian subjects. Conflicts of interest: The authors disclose no conflicts.
Applying Proteomics Integrated With Gene Microarray for Discovery of Colorectal Cancer Novel Biomarkers Xiao-fen Li,1 Jie-kai Yu,1 Wang-xiong Hu,1 and Shu Zheng1
Table 1. Components of a new index to predict advanced proximal neoplasia 1
1. Imperiale TF, Wagner DR, Lin CY, Larkin GN, Rogge JD, Ransohoff DF. Using Risk for Advanced Proximal Colonic Neoplasia to Tailor Endoscopic Screening for Colorectal Cancer. Ann Intern Med 2003; 139: 959-96510
Cancer Institute, the Second Affiliated Hospital, Zhejiang University School of Medicine, China BACKGROUND: Colorectal cancer (CRC) is the fourth leading
cause of cancer-related deaths worldwide. Proteins, as executives of genes’ instructions, are responsible for
e40
Abstracts
Clinical Gastroenterology and Hepatology Vol. 15, No. 1
cellular phenotypes. Integrating proteomics with gene microarray, we conducted this study to identify potential protein biomarkers of CRC. METHODS: Isobaric tags with related and absolute quantitation (iTRAQ) labeling mass spectrometry (MS) was applied to screen and identify differentially expressed proteins between paired CRC and adjacent normal mucosa. Meanwhile, Affymetrix U133plus2.0 microarrays were used to perform gene microarray analysis. Verification experiments included immunohistochemistry (IHC) and western blot of selected proteins. RESULTS: Overall, 5469 differentially expressed proteins were detected with iTRAQ-MS from 24 matched CRC and adjacent normal tissues. And gene microarray identified 39859 differential genes from 52 patients. Of these, 3083 differential proteins had corresponding differentially expressed genes, with 245 proteins and their genes showed >1.5-fold change in expression level. Gene ontology enrichment analysis revealed that up-regulated proteins were more involved in cell adhesion and motion than down-regulated proteins. In addition, up-regulated proteins were more likely to be located in nucleus and vesicles. Further verification experiments with IHC and western blot confirmed differential expression levels of 5 proteins (S100 calcium-binding protein A9, annexin A3, nicotinamide phosphoribosyltransferase, carboxylesterase 2 and calcium activated chloride channel A1) between CRC and normal tissues. CONCLUSION: S100 calcium-binding protein A9, annexin A3, nicotinamide phosphoribosyltransferase, carboxylesterase 2 and calcium activated chloride channel A1 were probably potential biomarkers of colorectal cancer. Annexin A3 may be associated with colorectal carcinogenesis.
of gastroduodenal mucosal injury would be assessed according to the modified Lanza Endoscopic Scoring System. RESULTS: From August 2015 to February 2016, 109 of 124 patients finished the follow-up, of whom 92.7% (101/ 109) underwent EGD. The prevalence of multiple gastroduodenal lesions, which should be treated with PPIs, is higher in ticagrelor plus aspirin group (38.5%, 15/39), compared with that in aspirin (33.3%, 10/30) or ticagrelor (31.3%, 10/32) monotherapy group, respectively. All patients were prescribed with PPIs for 1 month, 71.2% (72/101) of whom were treated with 3-month PPIs, only 7.0% (7/101) with more than 6-month. Among 17 patients with history of peptic ulcers, 17.6% (3/17) were treated with 1-month PPIs, 64.7% (11/17) with 3month, only 17.7% (3/17) with more than 6-month. CONCLUSION: It seems that Chinese patients are rarely under concomitant use of PPIs with antiplatelet therapy for more than 6 months, even those with history of peptic ulcer, demonstrating an unmet need for effective prophylactic treatment with PPIs to decrease the risk of upper GI bleeding.
Conflicts of interest: The authors disclose no conflicts.
Conflicts of interest: The authors disclose no conflicts.
Current Status of Concomitant Use of Proton Pump Inhibitors With Different Antiplatelet Therapies in Chinese Patients After Coronary Artery Bypass Grafting Chenyue Tang,1 Yunpeng Zhu,2 Cong Ye,2 Qiang Zhao,2 and Lifen Yu1 1
2
Department of Gastroenterology, and Cardiac Surgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, China BACKGROUND: Gastrointestinal complications of antiplate-
let drugs are common. Investigating the current status of concomitant use of proton pump inhibitors (PPIs) is needed in antiplatelet drug users after coronary artery bypass grafting (CABG). METHODS: Two hundred and seventy patients enrolled in Ruijin hospital, maximum proportion of those who were randomly assigned into three groups after CABG including aspirin or ticagrelor monotherapy group and ticagrelor plus aspirin group (DACAB trial, NCT 02201771), would undergo esophagogastroduodenoscopy (EGD) with their consent at 12th month. The grade
Capsule Endoscopy in Intestinal Lymphagiectasia: A Novel Endoscopic Classification and its Clinical Correlation Kui-liang Liu,1 Fang-yun Liu,1 Wen-bin Shen,2 and Jing Wu1 1
Department of Gastroenterology, Beijing Shijitan Hospital, Capital Medical University, and 2Department of Lymphatic Surgery, Beijing Shijitan Hospital, Capital Medical University BACKGROUND: Capsule Endoscopy (CE) is useful in diag-
nosis of intestinal lymphagiectasia (IL). However, systemic analysis of endoscopic and clinical characteristics remains unavailable. METHODS: Review the endoscopic characteristics of patients underwent CE in our endoscopy center between 2011 and 2015 and develop a novel endoscopic classification to investigate its clinical correlation. RESULTS: A total of 54 patients, 29 males and 25 females with median age of 24 (4-56) and duration lasting 20 days to 23 years were included. 42 cases were diagnosed as primary IL. Typical endoscopic characteristics included scattering, grouping or diffuse white-yellow