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Tu1944 Mutational Analysis of Pancreatic Neuroendocrine Tumor Using Endoscopic Ultrasound-Guided Fine-Needle Aspiration and Next-Generation Deep Sequencing
have no predictive value in local, advanced or metastatic PAC. This raises the question of the equivalence of the antibodies. Patients and Methods Murine (10D7G2) and rabbit (SP120) monoclonal anti-hENT1 antibodies were applied on the same tissue microarrays of resected PAC. Two sets of patients from 2 separate institutions (n=147 each) were used to compared hENT1 expression with both antibodies on two different immunochemistry automates. An additional amplification technique was performed on the second set. hENT1 expression was scored as previously reported. Results were correlated with overall survival (OS) following gemcitabine treatment. Results The rate of "hENT1 high" cases was lower with the SP120 clone (set 1: 7 versus 48%, set 2: 11 versus 38%). With the amplification technique, the rate of "hENT1 high" cases was globally similar between both antibodies. However, the concordance between the antibodies was found in only 50% of cases. High hENT1 expression was predictive of OS with the 10D7G2 antibody (HR=0.49; 95% CI: 0.24-0.98; P=0.045) but not with the SP120 (HR=1.61; 95% CI: 0.80-3.21; P=0.18). Conclusions: Our data suggest that the two antibodies are not equivalent. Further studies are ongoing and will be presented at the meeting: 1/ Comparative analysis of the rabbit antibody on a larger series (n=471) with staining performed by Ventana® (Roche®), 2/ Comparative analysis of 3 additional anti-hENT1 antibody together with mRNA level correlation, 3/ Comparative expression of hENT1 in a series of matched primary tumors-metastases.
Pancreas Cancer in Iran: Epidemiologic Feature, Risk Factors and Survival Ramin Shakeri, Akram Pourshams, Reza Tabrizi, Mehdi Mohamadnejad, Farhad Zamani, Ashraf Mohamadkhani, Sepideh Nikfam, Mohammad Reza Ostovaneh, Rasoul Sotoudehmanesh, Masoud Sotoudeh, Bijan Shahbazkhani, Farhad Islami, Farin Kamangar, Paolo Boffetta, Reza Malekzadeh Back ground and Aim: Pancreatic exocrine cancer (PC) is a deadly disease with a 5-year survival of less than 5%, and its etiology is largely unknown. We aimed to study risk factors, epidemiologic features, and survival of PC patients in Iran, using a case-control design. Methods: Cases and controls were selected from patients who were referred to four endoscopic ultrasound centers in Tehran, Iran. We recruited 307 histopathologically (n= 266, all adenocarcinoma) or clinically (n=41) diagnosed incident cases of PC, as well as 322 controls from those who had normal pancreas in enodosonography from January 2011 to March 2014. Results: Mean (±SD) age of cases and controls were 64.8 (±11.4) and 64.6 (±11.8) years and 180 (58.6%) cases and 144 (44.7%) of controls were male, respectively. Opium use was associated with an increased risk (OR 2.06; 95% CI 1.15-3.68) of pancreatic cancer. We did not find a significant association between ever tobacco smoking and PC risk. The median overall survival was 6.3 months and PC cases with pancreatic mass ≤ 3 cm, surgically respectable tumor, and ≥13 years of education had longer survival. Cases who were current opium user had a worse prognosis. Curative surgery was possible in only 16 cases (5.2%). Conclusion: PC presents at a very advanced stage, generally with an unsatisfactory outcome. This study suggests that opium use may be as an important risk factor of PC in Iran.
Tu1943 Can Metformin Change the Prognosis of Pancreatic Cancer? Sang Hyun Yoon, Hee Seung Lee, Moon Jae Chung, Jeong Youp Park, Seung Woo Park, Si Young Song, Jae Bock Chung, Seungmin Bang In recent studies, metformin has shown potential anti-cancer effects by lowering serum insulin and insulin-like growth factor 1 levels, and direct inhibition of cancer cell growth by activating 5' adenosine monophosphate-activated protein kinase protein, which inactivates proteins in the mammalian target of rapamycin pathway. The aim of this study was to evaluate potential anti-tumor effect of metformin on the treatment outcomes of patients with pancreatic adenocarcinoma (PAC). From May 2005 to December 2013, patients who were confirmed as PAC and diabetes mellitus (DM) were analyzed retrospectively. For this study, medical records including DM history, medication, clinical outcomes of PAC and radiologic images were reviewed. Exclusion criteria were insufficient medical records, no history of DM before diagnosis of PAC, no treatment of PAC or other than adenocarcinoma in histology. During the study period, 270 patients were included for the analysis. There were 175 patients (64.8%) with metformin exposure for DM management. The median overall survival time was 12.4 months for metformin-exposed group, and 8.8 months for metformin-unexposed group (P = 0.004, Log-rank test). In univariate analysis, metformin exposure, low serum CA19-9 level (<1000 U/mL), small primary cancer size (≤20mm), no tail involvement, good performance status and resectable cancer stage were associated with favorable outcome of survival. In multivariate analysis, metformin exposure, low serum CA19-9 level (<1000 U/mL) and resectable cancer stage were associated with favorable outcome. The treatment modality of DM other than metformin (insulin, sulfonylurea, thiazolidinedione, dipeptidyl peptidase IV inhibitor) did not show the significant effect on survival. Metformin exposure showed favorable treatment outcomes in PAC patients. Future prospective studies are required.
Tu1941 Opium Use and Risk of Pancreatic Cancer: A Prospective Cohort Study Mehdi Mohamadnejad, Akram Pourshams, Hossein Poustchi, Farhad Islami, maryam sharafkhah, Shirin Moossavi, Siavosh Nasseri-Moghaddam, Shahryar Semnani, Ramin Shakeri, Arash Etemadi, Shahin Merat, Masoud Khoshnia, Sanford M. Dawsey, Paul D. Pharoah, Paul Brennan, Christian C. Abnet, Paolo Boffetta, Farin Kamangar, Reza Malekzadeh Background: Opium use has been associated with higher risk of cancers of the esophagus, stomach, bladder, larynx, and lung; however its effect on the development of pancreatic cancer is unknown. Methods: We used data from the Golestan Cohort Study (GCS), a prospective cohort study in northeastern Iran with detailed, validated data on opium use and several other exposures. We examined the association of opium use and other risk factors with pancreatic cancer in 48,925 participants of GCS. The participants were followed up to 10 years with a follow up success rate of 99%. Pancreatic cancer cases and death were ascertained through local cancer and death registries, annual phone calls to study participants, and communicating with local health workers. Those with invasive cancers (other than non-melanoma skin cancer) were excluded from the control group. Patients with pancreatic cancer were diagnosed histologically, or based on positive clinical signs and symptoms, imaging findings and clinical follow up. Results: A median follow up of 7.14 years (344,517 person-years) resulted in 47 incident pancreatic cancer cases. On univariate analyses, age, opium use, cigarette smoking, body mass index (BMI), and marital status were associated with pancreatic cancer risk; whereas gender, alcohol use, socioeconomic status, or level of education were not (Table 1). The association of opium with pancreatic cancer was dose dependent, with a HR of 2.68 (1.28-5.60) for above median use compared to no use (P: 0.009). Due to collinearity between smoking and opium use, we generated a combined opium/smoking variable for multivariate analysis. In these analyses, combined smoking/opium use (hazard ratio, 3.00; 95% confidence interval, 1.40-6.42), being not currently married (hazard ratio, 2.14; 95% confidence interval, 1.07-4.29), and older age (hazard ratio for each year, 1.04; 95% confidence interval, 1.01-1.1) significantly increased risk of pancreatic cancer. When opium dose was entered the regression model as a continuous variable, the following factors were associated with pancreatic cancer: marital status (HR, 2.17; 95% confidence interval, 1.07-4.38), opium dose (HR, 1.002; 95% confidence interval, 1.0002-1.003), age (HR, 1.04; 95% confidence interval, 1.01-1.07), and smoking (HR, 2.17; 95% confidence interval, 1.14-4.14). Conclusion: Combined smoking and opium use, and marital status increase the risk of development of pancreatic cancer. Opium use is associated with pancreatic cancer in a dose-dependent manner. Univariate analysis of the association of factors with pancreatic cancer
Tu1944 Mutational Analysis of Pancreatic Neuroendocrine Tumor Using Endoscopic Ultrasound-Guided Fine-Needle Aspiration and Next-Generation Deep Sequencing Yoshimasa Kubota, Hiroshi Kawakami, Mitsuteru Natsuizaka, Masaki Kuwatani, Kazumichi Kawakubo, Yoko Abe, Shuhei Kawahata, Kimitoshi Kubo, Naoya Sakamoto Background: Pancreatic neuroendocrine tumor (PNET) is a rare neoplasm. Its prognosis varies within the same disease entity with median survival time of 136 months in localized cases and 24 months in metastastic cases. To date many attempts have been made to determine useful prognostic factors of PNET. Objectives: Genetical alteration of PNET was explored to determine the prognostic factor in linkage of clinical and genetical features. Methods: A total of 18 PNET cases were examined. All tumors were sporadic PNET. Tumor samples were obtained by either EUS-FNA (n = 16) or surgery (n = 2). Matched DNA from the peripheral blood (n = 3) or frozen leukocytes (n = 15) were used as controls. Samples were obtained at Hokkaido University Hospital, Sapporo, Japan, between January 2008 and September 2013. EUS-FNA was performed using a curvilinear echoendoscope and 22-gauge needles. The obtained specimen were stained using the rapid Romanowsky technique for quick interpretation and assessment of sample adequacy. We designed our original Ion AmpliSeqTM Custom Panels (Life Technologies) including 36 genes for the present study. Twenty nano grams of each genomic DNA samples and Ion AmpliseqTM Library Kits 2.0 (Life Technologies) were used for library preparation. The concentration and amplicon size of the libraries were determined by using an Agilent 2100 Bioanalyzer (Agilent Technologies). Ion OneTouchTM System (Life Technologies) was used for emulsion PCR. Samples were subsequently enriched using Ion OneTouchTM ES (Life Technologies). Sequencing was performed on an Ion PGMTM Sequencer by using an Ion 318TM chip (Life Technologies) with an Ion PGMTM Template OT2 200 kit (Life Technologies). Obtained sequences were mapped onto the human reference genome hg19, and variants were detected using Ion Torrent Suite v4.0.2 software (Life Technologies). Survival analyses were performed using Kaplan-Meier method and log-rank test for significance. Results: Of the 18 cases, 5 were classified as WHO grade NET G1, 8 as NET G2, 3 as NEC, and 2 as unknown. An average of 148 and 146 mutations were detected in 29 and 28 genes of tumors and controls, respectively. Tumor specific mutations in WHSC1 (28%, 5/18) were the most frequent, followed by DAXX/ATRX (22%, 4/18) and MLL3 (17%, 3/18). DAXX/ATRX mutations were only seen in NET G1 and G2, while KRAS and TP53 mutations were only seen in NEC. WHO grade (G1, G2 or NEC, p<0.01), exsistence of metastasis (p<0.05), mutations in MAPK signaling (KRAS, TP53, and DAXX, p<0.01), and mutations in pancreatic adenocarcinoma driver genes (KRAS, TP53, and SMAD4, p<0.05) were significantly associated with patients survivals. Conclusions: Mutational profiles of NET G1/G2 and NEC are completely different. Mutations in KRAS and TP53 are significantly associated with worse prognosis.
Tu1942 hENT1 Testing in Pancreatic Ductal Adenocarcinoma: Are We Ready yet? A Comparative Analysis of the Murine and the Rabbit Antibodies Jerome Cros, Raphaël Maréchal, Jean-Baptiste Bachet, Pierre Laurent-Puig, Jean Francois Flejou, Pieter Demetter, Magali Svrcek Introduction Human equilibrative nucleoside transporter 1 (hENT1) level of expression in pancreatic adenocarcinoma (PAC) may predict survival in gemcitabine-treated patients after resection. These results have been obtained with a murine anti-hENT1 antibody (10D7G2), which is not commercially available. Another antibody, rabbit-derived (SP120), appears to
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AGA Abstracts
AGA Abstracts
Tu1940
Pancreas Cancer in Iran: Epidemiologic Feature, Risk Factors and Survival Ramin Shakeri, Akram Pourshams, Reza Tabrizi, Mehdi Mohamadnejad, Farhad Zamani, Ashraf Mohamadkhani, Sepideh Nikfam, Mohammad Reza Ostovaneh, Rasoul Sotoudehmanesh, Masoud Sotoudeh, Bijan Shahbazkhani, Farhad Islami, Farin Kamangar, Paolo Boffetta, Reza Malekzadeh Back ground and Aim: Pancreatic exocrine cancer (PC) is a deadly disease with a 5-year survival of less than 5%, and its etiology is largely unknown. We aimed to study risk factors, epidemiologic features, and survival of PC patients in Iran, using a case-control design. Methods: Cases and controls were selected from patients who were referred to four endoscopic ultrasound centers in Tehran, Iran. We recruited 307 histopathologically (n= 266, all adenocarcinoma) or clinically (n=41) diagnosed incident cases of PC, as well as 322 controls from those who had normal pancreas in enodosonography from January 2011 to March 2014. Results: Mean (±SD) age of cases and controls were 64.8 (±11.4) and 64.6 (±11.8) years and 180 (58.6%) cases and 144 (44.7%) of controls were male, respectively. Opium use was associated with an increased risk (OR 2.06; 95% CI 1.15-3.68) of pancreatic cancer. We did not find a significant association between ever tobacco smoking and PC risk. The median overall survival was 6.3 months and PC cases with pancreatic mass ≤ 3 cm, surgically respectable tumor, and ≥13 years of education had longer survival. Cases who were current opium user had a worse prognosis. Curative surgery was possible in only 16 cases (5.2%). Conclusion: PC presents at a very advanced stage, generally with an unsatisfactory outcome. This study suggests that opium use may be as an important risk factor of PC in Iran.
Tu1943 Can Metformin Change the Prognosis of Pancreatic Cancer? Sang Hyun Yoon, Hee Seung Lee, Moon Jae Chung, Jeong Youp Park, Seung Woo Park, Si Young Song, Jae Bock Chung, Seungmin Bang In recent studies, metformin has shown potential anti-cancer effects by lowering serum insulin and insulin-like growth factor 1 levels, and direct inhibition of cancer cell growth by activating 5' adenosine monophosphate-activated protein kinase protein, which inactivates proteins in the mammalian target of rapamycin pathway. The aim of this study was to evaluate potential anti-tumor effect of metformin on the treatment outcomes of patients with pancreatic adenocarcinoma (PAC). From May 2005 to December 2013, patients who were confirmed as PAC and diabetes mellitus (DM) were analyzed retrospectively. For this study, medical records including DM history, medication, clinical outcomes of PAC and radiologic images were reviewed. Exclusion criteria were insufficient medical records, no history of DM before diagnosis of PAC, no treatment of PAC or other than adenocarcinoma in histology. During the study period, 270 patients were included for the analysis. There were 175 patients (64.8%) with metformin exposure for DM management. The median overall survival time was 12.4 months for metformin-exposed group, and 8.8 months for metformin-unexposed group (P = 0.004, Log-rank test). In univariate analysis, metformin exposure, low serum CA19-9 level (<1000 U/mL), small primary cancer size (≤20mm), no tail involvement, good performance status and resectable cancer stage were associated with favorable outcome of survival. In multivariate analysis, metformin exposure, low serum CA19-9 level (<1000 U/mL) and resectable cancer stage were associated with favorable outcome. The treatment modality of DM other than metformin (insulin, sulfonylurea, thiazolidinedione, dipeptidyl peptidase IV inhibitor) did not show the significant effect on survival. Metformin exposure showed favorable treatment outcomes in PAC patients. Future prospective studies are required.
Tu1941 Opium Use and Risk of Pancreatic Cancer: A Prospective Cohort Study Mehdi Mohamadnejad, Akram Pourshams, Hossein Poustchi, Farhad Islami, maryam sharafkhah, Shirin Moossavi, Siavosh Nasseri-Moghaddam, Shahryar Semnani, Ramin Shakeri, Arash Etemadi, Shahin Merat, Masoud Khoshnia, Sanford M. Dawsey, Paul D. Pharoah, Paul Brennan, Christian C. Abnet, Paolo Boffetta, Farin Kamangar, Reza Malekzadeh Background: Opium use has been associated with higher risk of cancers of the esophagus, stomach, bladder, larynx, and lung; however its effect on the development of pancreatic cancer is unknown. Methods: We used data from the Golestan Cohort Study (GCS), a prospective cohort study in northeastern Iran with detailed, validated data on opium use and several other exposures. We examined the association of opium use and other risk factors with pancreatic cancer in 48,925 participants of GCS. The participants were followed up to 10 years with a follow up success rate of 99%. Pancreatic cancer cases and death were ascertained through local cancer and death registries, annual phone calls to study participants, and communicating with local health workers. Those with invasive cancers (other than non-melanoma skin cancer) were excluded from the control group. Patients with pancreatic cancer were diagnosed histologically, or based on positive clinical signs and symptoms, imaging findings and clinical follow up. Results: A median follow up of 7.14 years (344,517 person-years) resulted in 47 incident pancreatic cancer cases. On univariate analyses, age, opium use, cigarette smoking, body mass index (BMI), and marital status were associated with pancreatic cancer risk; whereas gender, alcohol use, socioeconomic status, or level of education were not (Table 1). The association of opium with pancreatic cancer was dose dependent, with a HR of 2.68 (1.28-5.60) for above median use compared to no use (P: 0.009). Due to collinearity between smoking and opium use, we generated a combined opium/smoking variable for multivariate analysis. In these analyses, combined smoking/opium use (hazard ratio, 3.00; 95% confidence interval, 1.40-6.42), being not currently married (hazard ratio, 2.14; 95% confidence interval, 1.07-4.29), and older age (hazard ratio for each year, 1.04; 95% confidence interval, 1.01-1.1) significantly increased risk of pancreatic cancer. When opium dose was entered the regression model as a continuous variable, the following factors were associated with pancreatic cancer: marital status (HR, 2.17; 95% confidence interval, 1.07-4.38), opium dose (HR, 1.002; 95% confidence interval, 1.0002-1.003), age (HR, 1.04; 95% confidence interval, 1.01-1.07), and smoking (HR, 2.17; 95% confidence interval, 1.14-4.14). Conclusion: Combined smoking and opium use, and marital status increase the risk of development of pancreatic cancer. Opium use is associated with pancreatic cancer in a dose-dependent manner. Univariate analysis of the association of factors with pancreatic cancer
Tu1944 Mutational Analysis of Pancreatic Neuroendocrine Tumor Using Endoscopic Ultrasound-Guided Fine-Needle Aspiration and Next-Generation Deep Sequencing Yoshimasa Kubota, Hiroshi Kawakami, Mitsuteru Natsuizaka, Masaki Kuwatani, Kazumichi Kawakubo, Yoko Abe, Shuhei Kawahata, Kimitoshi Kubo, Naoya Sakamoto Background: Pancreatic neuroendocrine tumor (PNET) is a rare neoplasm. Its prognosis varies within the same disease entity with median survival time of 136 months in localized cases and 24 months in metastastic cases. To date many attempts have been made to determine useful prognostic factors of PNET. Objectives: Genetical alteration of PNET was explored to determine the prognostic factor in linkage of clinical and genetical features. Methods: A total of 18 PNET cases were examined. All tumors were sporadic PNET. Tumor samples were obtained by either EUS-FNA (n = 16) or surgery (n = 2). Matched DNA from the peripheral blood (n = 3) or frozen leukocytes (n = 15) were used as controls. Samples were obtained at Hokkaido University Hospital, Sapporo, Japan, between January 2008 and September 2013. EUS-FNA was performed using a curvilinear echoendoscope and 22-gauge needles. The obtained specimen were stained using the rapid Romanowsky technique for quick interpretation and assessment of sample adequacy. We designed our original Ion AmpliSeqTM Custom Panels (Life Technologies) including 36 genes for the present study. Twenty nano grams of each genomic DNA samples and Ion AmpliseqTM Library Kits 2.0 (Life Technologies) were used for library preparation. The concentration and amplicon size of the libraries were determined by using an Agilent 2100 Bioanalyzer (Agilent Technologies). Ion OneTouchTM System (Life Technologies) was used for emulsion PCR. Samples were subsequently enriched using Ion OneTouchTM ES (Life Technologies). Sequencing was performed on an Ion PGMTM Sequencer by using an Ion 318TM chip (Life Technologies) with an Ion PGMTM Template OT2 200 kit (Life Technologies). Obtained sequences were mapped onto the human reference genome hg19, and variants were detected using Ion Torrent Suite v4.0.2 software (Life Technologies). Survival analyses were performed using Kaplan-Meier method and log-rank test for significance. Results: Of the 18 cases, 5 were classified as WHO grade NET G1, 8 as NET G2, 3 as NEC, and 2 as unknown. An average of 148 and 146 mutations were detected in 29 and 28 genes of tumors and controls, respectively. Tumor specific mutations in WHSC1 (28%, 5/18) were the most frequent, followed by DAXX/ATRX (22%, 4/18) and MLL3 (17%, 3/18). DAXX/ATRX mutations were only seen in NET G1 and G2, while KRAS and TP53 mutations were only seen in NEC. WHO grade (G1, G2 or NEC, p<0.01), exsistence of metastasis (p<0.05), mutations in MAPK signaling (KRAS, TP53, and DAXX, p<0.01), and mutations in pancreatic adenocarcinoma driver genes (KRAS, TP53, and SMAD4, p<0.05) were significantly associated with patients survivals. Conclusions: Mutational profiles of NET G1/G2 and NEC are completely different. Mutations in KRAS and TP53 are significantly associated with worse prognosis.
Tu1942 hENT1 Testing in Pancreatic Ductal Adenocarcinoma: Are We Ready yet? A Comparative Analysis of the Murine and the Rabbit Antibodies Jerome Cros, Raphaël Maréchal, Jean-Baptiste Bachet, Pierre Laurent-Puig, Jean Francois Flejou, Pieter Demetter, Magali Svrcek Introduction Human equilibrative nucleoside transporter 1 (hENT1) level of expression in pancreatic adenocarcinoma (PAC) may predict survival in gemcitabine-treated patients after resection. These results have been obtained with a murine anti-hENT1 antibody (10D7G2), which is not commercially available. Another antibody, rabbit-derived (SP120), appears to
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AGA Abstracts
AGA Abstracts
Tu1940