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Analysis of immunological features in blood of patients with pancreatic ductal adenocarcinoma
S116
Abstracts / Pancreatology 16 (2016) S1eS192
candidate phosphoproteins in PDAC tissues by immunohistochemistry. Next, a specific ANX2 enzyme-linked immunosorbent assay (ELISA) was originally established to measure the levels of soluble ANX2 in serum. We analyzed the correlations between serum ANX2 levels and clinico-pathological characteristics, recurrence form after surgery in PDAC patients. Results: The comprehensive analyses showed up-regulation of p-Akt in GEM-MIA PaCa-2 cells in which ANX2 is highly expressed. The expression level of p-Akt was significantly correlated with that of the downstream protein, p-mTOR, in PDAC tissues. Inhibition of mTOR phosphorylation canceled gemcitabine resistance in GEM-MIA PaCa-2 cells. The levels of Anx2 were significantly higher in the preoperative serum than in the postoperative serum in PDAC patients (p＜0.02). Kaplan Meier analyses showed that the high level of Anx2 in preoperative serum significantly correlated with rapid recurrence (p＜0.02). These results indicate that circulating ANX2 may accelerate cancer progression in PDAC, resulting in rapid recurrence of PDAC patients after gemcitabine-based adjuvant chemotherapy. Conclusions: The level of Anx2 inducing gemcitabine resistance in sera of patients may predict its clinical outcome, leading to the development of a novel strategy for tailor-made treatment for PDAC.
P-078. Overexpression of KIAA1199 predicts poor prognosis in pancreatic cancer Atsuhiro Koga, Norihiro Sato, Shiro Kohi, Xiao-Bo Cheng, Keiji Hirata University of Occupational and Environmental Health, Japan Background: Progression of cancer is often associated with interactions between cancer cells and extracellular matrices including hyaluronan (HA). In addition to increased production of HA, degradation of HA has been shown to play a critical role in cancer invasion and metastasis. Recently, a novel HA-degrading enzyme, KIAA1199, has been identified as overexpressed in various cancers. In the present study, we investigated the expression of KIAA1199 and its clinicopathological correlations in pancreatic ductal adenocarcinoma (PDAC). Methods: We used real time RT-PCR to examine mRNA expression of KIAA1199 in frozen tissues from 14 patients with PDAC. We also performed immunohistochemistry to determine the expression pattern of KIAA1199 in 98 patients with PDAC. The expression pattern of KIAA1199 was correlated with clinicopathological variables, including survival time after surgical resection, using univariate and multivariate analyses. Results: KIAA1199 mRNA expression was higher in all PDAC tissues as compared to the corresponding non-tumoral pancreatic tissues (P¼2.24E6). Immunohistochemical staining revealed strong expression of KIAA1199 in 32 patients (32.7%). The overall survival was significantly shorter in patients with strong KIAA expression than in patients with low or no KIAA1199 expression (median, 12.3 months versus 27.1 months, P¼0.0003). In multivariate analysis, strong KIAA1199 expression (P¼0.01), UICC Stage (P¼0.019), and residual tumor (P¼0.029) were independent factors predicting poor prognosis in PDAC patients. Conclusion: Our findings suggest that KIAA1199 can be used as a prognostic marker in patients with PDAC.
P-079. Analysis of immunological features in blood of patients with pancreatic ductal adenocarcinoma Takuya Komura 1, Yoshio Sakai 2, 3, Hisashi Takabatake 1, Norihiko Ogawa 1, Masaki Miyazawa 1, Shuichi Kaneko 1, 2 1
Disease Control and Homeostasis, Kanazawa University, Japan Department of Gastroenterology, Kanazawa University, Japan 3 Department of Laboratory Medicine, Kanazawa University, Japan 2
Pancreatic ductal adenocarcinoma (PDAC) is an extremely aggressive malignancy with a poor prognosis. Immune system of the host significantly affects the progression as well as responsiveness of treatment for cancer, including PDAC. Therefore, it is extremely important to understand the host inflammatory immune response of PDAC for establishment of the novel diagnostic method to detect PDAC as well as development of therapy. We examined cytokines/chemokines concentration in sera of 50 PDAC patients. Concentrations of IL-6, IL-7, IL-15, MCP-1, and IP-10 in sera of PDAC patients were significantly elevated, compared with those of 27 healthy volunteers. As for these elevated cytokines/chemokines in PDAC patients, we measured mRNAs expression in CD4+ T cells and CD14+ monocytes by real-time detection PCR (RTD-PCR). We observed IL-15 expression by CD14+ monocytes, and IL-6 and IL-7 expression by CD4+ T cells of PDAC patients were significantly up-regulated, respectively, suggesting that CD4+ T cells and CD14+ monocytes presumably contribute to increment of these cytokines/chemokines concentrations in sera of PDAC patients. We also examined how gene expressionfeatures of CD4+T cell and CD14+ monocyte fractions in blood of PDAC patients using DNA microarray. The biological processes of altered genes in CD4+ T cells of PDAC patients were related to the cell cycle and inflammation as well as DNA damage and apoptosis. The biological process networks altered genes in CD14+ monocytes of PDAC patients were related to the cell cycle, inflammation, blood coagulation, cell adhesion, and development. Furthermore, we observed that gene expression levels related to immune checkpoint molecules, PD-1, was increased, whereas CTLA-4, which is important for inhibitory molecule for immune reaction, was not increased, in CD4+T cells of PDAC patients using RTD-PCR. In conclusion, this study demonstrated that immune response reflected in blood of PDAC patients is highlighted the presence of activated CD4+ T cells and CD14+ monocytes.
P-080. The analysis of circulating tumor DNA in patients with resectable pancreatic cancer Naoto Hadano 1, Yoshiaki Murakami 1, Kenichiro Uemura 1, Naru Kondo 1, Naoya Nakagawa 1, Taijiro Sueda 1, Eiso Hiyama 2 1 Department of Surgery, Applied Life Sciences Institute of Biomedical and Health Sciences, Hiroshima University, Japan 2 Natural Science Center for Basic Research and Development (NBARD), Hiroshima University, Japan
Background: Pancreatic ductal adenocarcinoma (PDAC) is frequently diagnosed at an advanced stage, leading to a poor prognosis. Therefore, interest in the development of noninvasive biomarkers for prognostic prediction has grown rapidly. Here, we assessed the clinical implications of v-Ki-ras2 kirsten rat sarcoma viral oncogene homolog (KRAS)-mutated circulating tumour DNA (ctDNA) as a useful surrogate biomarker in patients with PDAC. Methods: We used droplet digital polymerase chain reaction to detect rare mutant tumour-derived KRAS genes in plasma cell-free DNA (cfDNA) as ctDNA. Samples were collected from 105 patients who underwent pancreatic resection for PDAC at a single institution. Overall survival (OS) was analysed according to the presence of ctDNA. Results: Among the 105 cases, ctDNA was detected in 33 (31%) plasma samples. The median OS durations were 13.6 months for patients with ctDNA (ctDNA+) and 27.6 months for patients without ctDNA. ctDNA+ patients had a significantly poorer prognosis with respect to OS (P＜ 0.0001). Conclusions: Our findings suggested that the presence of ctDNA in plasma samples could be an important and powerful predictor of poor survival in patients with PDACs. Accordingly, ctDNA detection may be a promising approach with respect to PDAC treatment.
Abstracts / Pancreatology 16 (2016) S1eS192
candidate phosphoproteins in PDAC tissues by immunohistochemistry. Next, a specific ANX2 enzyme-linked immunosorbent assay (ELISA) was originally established to measure the levels of soluble ANX2 in serum. We analyzed the correlations between serum ANX2 levels and clinico-pathological characteristics, recurrence form after surgery in PDAC patients. Results: The comprehensive analyses showed up-regulation of p-Akt in GEM-MIA PaCa-2 cells in which ANX2 is highly expressed. The expression level of p-Akt was significantly correlated with that of the downstream protein, p-mTOR, in PDAC tissues. Inhibition of mTOR phosphorylation canceled gemcitabine resistance in GEM-MIA PaCa-2 cells. The levels of Anx2 were significantly higher in the preoperative serum than in the postoperative serum in PDAC patients (p＜0.02). Kaplan Meier analyses showed that the high level of Anx2 in preoperative serum significantly correlated with rapid recurrence (p＜0.02). These results indicate that circulating ANX2 may accelerate cancer progression in PDAC, resulting in rapid recurrence of PDAC patients after gemcitabine-based adjuvant chemotherapy. Conclusions: The level of Anx2 inducing gemcitabine resistance in sera of patients may predict its clinical outcome, leading to the development of a novel strategy for tailor-made treatment for PDAC.
P-078. Overexpression of KIAA1199 predicts poor prognosis in pancreatic cancer Atsuhiro Koga, Norihiro Sato, Shiro Kohi, Xiao-Bo Cheng, Keiji Hirata University of Occupational and Environmental Health, Japan Background: Progression of cancer is often associated with interactions between cancer cells and extracellular matrices including hyaluronan (HA). In addition to increased production of HA, degradation of HA has been shown to play a critical role in cancer invasion and metastasis. Recently, a novel HA-degrading enzyme, KIAA1199, has been identified as overexpressed in various cancers. In the present study, we investigated the expression of KIAA1199 and its clinicopathological correlations in pancreatic ductal adenocarcinoma (PDAC). Methods: We used real time RT-PCR to examine mRNA expression of KIAA1199 in frozen tissues from 14 patients with PDAC. We also performed immunohistochemistry to determine the expression pattern of KIAA1199 in 98 patients with PDAC. The expression pattern of KIAA1199 was correlated with clinicopathological variables, including survival time after surgical resection, using univariate and multivariate analyses. Results: KIAA1199 mRNA expression was higher in all PDAC tissues as compared to the corresponding non-tumoral pancreatic tissues (P¼2.24E6). Immunohistochemical staining revealed strong expression of KIAA1199 in 32 patients (32.7%). The overall survival was significantly shorter in patients with strong KIAA expression than in patients with low or no KIAA1199 expression (median, 12.3 months versus 27.1 months, P¼0.0003). In multivariate analysis, strong KIAA1199 expression (P¼0.01), UICC Stage (P¼0.019), and residual tumor (P¼0.029) were independent factors predicting poor prognosis in PDAC patients. Conclusion: Our findings suggest that KIAA1199 can be used as a prognostic marker in patients with PDAC.
P-079. Analysis of immunological features in blood of patients with pancreatic ductal adenocarcinoma Takuya Komura 1, Yoshio Sakai 2, 3, Hisashi Takabatake 1, Norihiko Ogawa 1, Masaki Miyazawa 1, Shuichi Kaneko 1, 2 1
Disease Control and Homeostasis, Kanazawa University, Japan Department of Gastroenterology, Kanazawa University, Japan 3 Department of Laboratory Medicine, Kanazawa University, Japan 2
Pancreatic ductal adenocarcinoma (PDAC) is an extremely aggressive malignancy with a poor prognosis. Immune system of the host significantly affects the progression as well as responsiveness of treatment for cancer, including PDAC. Therefore, it is extremely important to understand the host inflammatory immune response of PDAC for establishment of the novel diagnostic method to detect PDAC as well as development of therapy. We examined cytokines/chemokines concentration in sera of 50 PDAC patients. Concentrations of IL-6, IL-7, IL-15, MCP-1, and IP-10 in sera of PDAC patients were significantly elevated, compared with those of 27 healthy volunteers. As for these elevated cytokines/chemokines in PDAC patients, we measured mRNAs expression in CD4+ T cells and CD14+ monocytes by real-time detection PCR (RTD-PCR). We observed IL-15 expression by CD14+ monocytes, and IL-6 and IL-7 expression by CD4+ T cells of PDAC patients were significantly up-regulated, respectively, suggesting that CD4+ T cells and CD14+ monocytes presumably contribute to increment of these cytokines/chemokines concentrations in sera of PDAC patients. We also examined how gene expressionfeatures of CD4+T cell and CD14+ monocyte fractions in blood of PDAC patients using DNA microarray. The biological processes of altered genes in CD4+ T cells of PDAC patients were related to the cell cycle and inflammation as well as DNA damage and apoptosis. The biological process networks altered genes in CD14+ monocytes of PDAC patients were related to the cell cycle, inflammation, blood coagulation, cell adhesion, and development. Furthermore, we observed that gene expression levels related to immune checkpoint molecules, PD-1, was increased, whereas CTLA-4, which is important for inhibitory molecule for immune reaction, was not increased, in CD4+T cells of PDAC patients using RTD-PCR. In conclusion, this study demonstrated that immune response reflected in blood of PDAC patients is highlighted the presence of activated CD4+ T cells and CD14+ monocytes.
P-080. The analysis of circulating tumor DNA in patients with resectable pancreatic cancer Naoto Hadano 1, Yoshiaki Murakami 1, Kenichiro Uemura 1, Naru Kondo 1, Naoya Nakagawa 1, Taijiro Sueda 1, Eiso Hiyama 2 1 Department of Surgery, Applied Life Sciences Institute of Biomedical and Health Sciences, Hiroshima University, Japan 2 Natural Science Center for Basic Research and Development (NBARD), Hiroshima University, Japan
Background: Pancreatic ductal adenocarcinoma (PDAC) is frequently diagnosed at an advanced stage, leading to a poor prognosis. Therefore, interest in the development of noninvasive biomarkers for prognostic prediction has grown rapidly. Here, we assessed the clinical implications of v-Ki-ras2 kirsten rat sarcoma viral oncogene homolog (KRAS)-mutated circulating tumour DNA (ctDNA) as a useful surrogate biomarker in patients with PDAC. Methods: We used droplet digital polymerase chain reaction to detect rare mutant tumour-derived KRAS genes in plasma cell-free DNA (cfDNA) as ctDNA. Samples were collected from 105 patients who underwent pancreatic resection for PDAC at a single institution. Overall survival (OS) was analysed according to the presence of ctDNA. Results: Among the 105 cases, ctDNA was detected in 33 (31%) plasma samples. The median OS durations were 13.6 months for patients with ctDNA (ctDNA+) and 27.6 months for patients without ctDNA. ctDNA+ patients had a significantly poorer prognosis with respect to OS (P＜ 0.0001). Conclusions: Our findings suggested that the presence of ctDNA in plasma samples could be an important and powerful predictor of poor survival in patients with PDACs. Accordingly, ctDNA detection may be a promising approach with respect to PDAC treatment.