Poor Prognosis In Pancreatic Cancer Research Articles

Abstract 2289: Evaluation of novel genetically engineered anti-MUC16 antibody for therapeutic targeting in pancreatic cancer

Abstract Introduction: MUC16, the largest membrane glycoprotein, is not only important for protection under normal physiological conditions, but also contributes to disease progression and metastasis in various malignancies. TCGA analysis along with clinicopathologic data validate the association of aberrant MUC16 overexpression with poor prognosis in pancreatic cancer. Research from different groups and our lab has demonstrated the contribution of surface tethered MUC16 in tumorigenesis and metastasis. Further, in a recent study MUC16 was identified as a neoantigen associated with long-term pancreatic cancer survivors, which further supports its utility as a therapeutic target. With this in mind, we have recently generated a panel of monoclonal antibodies against tumorigenic carboxy terminal (CT) portion of MUC16. Method: We characterized anti-MUC16 mAbs in different pancreatic cancer cell lines and selected mAb5E6 for further engineering as therapeutic entity. We constructed novel chimeric mAb by grafting mouse heavy and light chain variable fragment (Fv) region on constant portion of human heavy (FcH ) and light (FcL) chain under IL-2 secretion system followed by their co transfection in CHO-K1 cells. These chimeric antibodies were characterized for their binding in different assays and evaluated for functional activity. Results and summary: Anti-MUC16 mAb 5E6 recognized different molecular weight forms of MUC16 in several pancreatic cancer cell lines and successfully inhibited the invasion (45%) and migration (62%) of MUC16 CT transfected MiaPaca cells in in vitro assays. Therefore, we engineered this antibody for therapeutic targeting. Chimeric antibody purified from culture supernatant recognized both recombinant and endogenous MUC16 in different binding assays. Interestingly, chimeric mAb 5E6 displayed better affinity (50nM) than its parent murine version (478nM) and recognized different forms of endogenous MUC16. Chimeric mAb5E6 decreased the colony formation ability (50%) and migration (~40%) of MUC16 expressing pancreatic cancer cell lines. Besides, chimeric mAb 5E6 treated pancreatic cancer cells showed reduction in FAK and Akt signaling associated with tumorigenesis and migration. Of importance, the chimeric antibody was internalized by MUC16 expressing pancreatic cancer cells and localized in the lysosomes in both time and temperature dependent manner which suggest their implication for payload delivery. Conclusions: Genetically engineered anti-MUC16 antibody decreased the tumorigenicity of pancreatic cancer cells. Our data validated testing the novel anti-MUC16 chimeric antibody for payload delivery and therapeutic targeting in pancreatic cancer. Citation Format: Ashu Shah, Abhijit Aithal, Catherine Orzechowski, Saravanakumar Marimuthu, Prakash Kshirsagar, Moorthy P. Ponnusamy, Maneesh Jain, Surinder K. Batra. Evaluation of novel genetically engineered anti-MUC16 antibody for therapeutic targeting in pancreatic cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2289.

A novel antisense lncRNA NT5E promotes progression by modulating the expression of SYNCRIP and predicts a poor prognosis in pancreatic cancer

A novel antisense lncRNA NT5E was identified in a previous microarray that was clearly up‐regulated in pancreatic cancer (PC) tissues. However, its biological function remains unclear. Thus, we aimed to explore its function and clinical significance in PC. The lncNT5E expression was determined in PC specimens and cell lines. In vitro and in vivo studies detected the impact of lncNT5E depletion on PC cell proliferation, migration and invasion. Western blotting investigated the epithelial‐mesenchymal transition (EMT) markers. The interaction between lncNT5E and the promoter region of SYNCRIP was detected by dual‐luciferase reporter assay. The role of lncNT5E in modulating SYNCRIP was investigated in vitro. Our results showed that lncNT5E was significantly up‐regulated in PC tissues and cell lines and associated with poor prognosis. LncNT5E depletion inhibited PC cell proliferation, migration, invasion and EMT in vitro and caused tumorigenesis arrest in vivo. Furthermore, SYNCRIP knockdown had effects similar to those of lncNT5E depletion. A significant positive relationship was observed between lncNT5E and SYNCRIP. Moreover, the dual‐luciferase reporter assays indicated that lncNT5E depletion significantly inhibited SYNCRIP promoter activity. Importantly, the malignant phenotypes of lncNT5E depletion were rescued by overexpressing SYNCRIP. In conclusion, lncNT5E predicts poor prognosis and promotes PC progression by modulating SYNCRIP expression.

MicroRNA-7 as a Potential Biomarker for Prognosis in Pancreatic Cancer.

MicroRNAs play critical roles in tumor progression. Our recent study has indicated that microRNA-7 (miR-7) impairs autophagy-derived pools of glucose to suppress the glycolysis in pancreatic cancer progression. However, the roles of miR-7 in clinical significance and chemoresistance of pancreatic cancer remain unexplored. The aim of this study was to assess the expression of miR-7 in patients with pancreatic cancer and to evaluate the possibility of its usage as a prognostic molecular biomarker. MicroRNA array-based quantification analysis of 372 miRNAs was compared in serum between pancreatic cancer and healthy individuals, gemcitabine-sensitive and gemcitabine-resistance patients. We identified miR-7 showed the potential predictive power for gemcitabine-sensitive patients with pancreatic cancer. Then, the results were validated in pancreatic tissue microarray and The Cancer Genome Atlas (TCGA) dataset, demonstrating that lower miR-7 expression was correlated with more advanced tumor stages and worse prognosis in pancreatic cancer. The Cox proportional-hazards model analysis identified miR-7 to be an independent variable for prediction of the survival. Furthermore, the mechanistic exploration suggested the clinical significance of miR-7 involved its interference effect on autophagy and glycolysis in pancreatic cancer using pancreatic cancer tissue microarrays and TCGA data. Therefore, the results of the present study provide evidences that low microRNA-7 expression may contribute to tumor progression and poor prognosis in pancreatic cancer.

LncRNA HCP5 Regulates Pancreatic Cancer Progression by miR-140-5p/CDK8 Axis

Background: Pancreatic cancer (PC) is a leading cause of cancer-related deaths worldwide. Human leukocyte antigen complex P5 (HCP5), a member of long noncoding RNAs (lncRNAs), was reported to be associated with the poor prognosis of PC. However, the mechanism of HCP5 in regulating the progression of PC remains poorly defined. Materials and Methods: Quantitative real-time polymerase chain reaction was performed to detect the expression levels of HCP5, microRNA (miR)-140-5p, and cyclin-dependent kinase 8 (CDK8) in PC tissues and cells. Cell counting kit-8 (CCK-8) assay was utilized to check cell proliferation. Transwell assay was employed to evaluate the abilities of cell migration and invasion. Xenograft tumor model was established to investigate the biological role of HCP5 in PC in vivo. The interaction between miR-140-5p and HCP5 or CDK8 was predicted by starBase or TargetScan, respectively. The dual-luciferase reporter assay was conducted to corroborate the interaction. The protein level of CDK8 was measured by Western blot. Results: HCP5 and CDK8 were significantly upregulated in PC tissues and cells, opposite to the expression of miR-140-5p. High expression of HCP5 contributed to the low survival rate and HCP5 silencing inhibited proliferation, migration, and invasion of PC cells in vitro. Simultaneously, in vivo experiments indicated that downregulation of HCP5 suppressed tumor growth. In addition, miR-140-5p was a target of HCP5 and bound to the 3'-untranslated region (3'UTR) of CDK8. Further studies revealed that overexpression of CDK8 reversed the miR-140-5p-mediated inhibitory effect on PC progression. Moreover, downregulation of miR-140-5p or upregulation of CDK8 inverted the silencing-mediated repressive impact of HCP5 on PC progression. Conclusion: Downregulation of HCP5 impeded PC progression by downregulating CDK8 via sponging miR-140-5p.

High expression levels of polymeric immunoglobulin receptor are correlated with chemoresistance and poor prognosis in pancreatic cancer.

Pancreatic cancer has extremely poor prognosis, warranting the discovery of novel therapeutic and prognostic markers. The expression of polymeric immunoglobulin receptor (pIgR), a key component of the mucosal immune system, is increased in several cancers. However, its clinical relevance in pancreatic cancer remains unclear. In the present study, the prognostic value of pIgR in pancreatic cancer patients after surgical resection was assessed and it was determined that the expression of pIgR was correlated with poor prognosis. Ten pancreatic cancer patient-derived xenograft (PDX) lines were established, followed by next-generation sequencing of tumor tissues from these lines after standard chemotherapy. Immunohistochemical analysis of chemoresistance-related molecules using 77 pancreatic cancer tissues was also performed. The expression of pIgR mRNA in the PDX group treated with anticancer drugs was higher than in the untreated group. High pIgR expression in tissue specimens from 77 pancreatic cancer patients was significantly associated with poor prognosis and was revealed to be an independent prognostic factor, predicting poor outcomes. High pIgR mRNA and protein levels were independent prognostic factors, indicating that pIgR could be a novel predictor for poor prognosis of pancreatic cancer patients.

MiR-93 is related to poor prognosis in pancreatic cancer and promotes tumor progression by targeting microtubule dynamics

Biomarkers and effective therapeutic agents to improve the dismal prognosis of pancreatic ductal adenocarcinoma (PDAC) are urgently required. We aimed to analyze the prognostic value and mechanistic action of miR-93 in PDAC. Correlation of miR-93 tumor levels from 83 PDAC patients and overall survival (OS) was analyzed by Kaplan–Meier. MiR-93 depletion in PANC-1 and MIA PaCa-2 cells was achieved by CRISPR/Cas9 and miR-93 overexpression in HPDE cells by retroviral transduction. Cell proliferation, migration and invasion, cell cycle analysis, and in vivo tumor xenografts in nude mice were assessed. Proteomic analysis by mass spectrometry and western-blot was also performed. Finally, miR-93 direct binding to candidate mRNA targets was evaluated by luciferase reporter assays. High miR-93 tumor levels are significantly correlated with a worst prognosis in PDAC patients. MiR-93 abolition altered pancreatic cancer cells phenotype inducing a significant increase in cell size and a significant decrease in cell invasion and proliferation accompanied by a G2/M arrest. In vivo, lack of miR-93 significantly impaired xenograft tumor growth. Conversely, miR-93 overexpression induced a pro-tumorigenic behavior by significantly increasing cell proliferation, migration, and invasion. Proteomic analysis unveiled a large group of deregulated proteins, mainly related to G2/M phase, microtubule dynamics, and cytoskeletal remodeling. CRMP2, MAPRE1, and YES1 were confirmed as direct targets of miR-93. MiR-93 exerts oncogenic functions by targeting multiple genes involved in microtubule dynamics at different levels, thus affecting the normal cell division rate. MiR-93 or its direct targets (CRMP2, MAPRE1, or YES1) are new potential therapeutic targets for PDAC.

Research progression in neural invasion model of pancreatic cancer

Pancreatic cancer is a malignant tumor of the digestive system with occult onset, low early diagnosis rate, rapid progression and poor prognosis. Perineural invasion (PNI) of pancreatic cancer is considered as an important factor which leads to the poor prognosis of pancreatic cancer. Neural invasion models of pancreatic cancer can simulate the occurrence and development of neural invasion of pancreatic cancer under the complex tumor microenvironment, which is an important carrier to study the molecular mechanism, early diagnosis and treatment improvement of PNI of pancreatic cancer. The current PNI model could be broadly divided into in vivo model and in vitro model, and in vivo model could further be divided into ectopic model and in situ model. Recently, based on the in vivo and in vitro models, genetically engineered mouse models and patient-derived xenograft models have been applied. Here, the preparation methods, clinical uses and limitations of the commonly used neural invasion in vitro and in vivo models of pancreatic cancer are reviewed.

PLS3 predicts poor prognosis in pancreatic cancer and promotes cancer cell proliferation via PI3K/AKT signaling.

Plastin-3 plays a key role in cancer cell proliferation and invasion, but its prognostic value in pancreatic cancer (PACA) remains poorly defined. In this study, we show that PLS3 messenger RNA is overexpressed in PACA tissue compared with normal tissue. We accumulated 207 cases of PACA specimens to perform immunohistochemical analysis and demonstrated that PLS3 levels correlate with T-classification (p < .001) and pathology (p < .001). Furthermore, overall survival rates (p < .001) in tumors with high PLS3 expression were poor, as assessed through Kaplan-Meier survival analysis. PLS3 was found to be an independent prognostic factor for PACA through multivariate Cox regression analysis. Moreover, we found that PLS3 enhances the proliferation and invasion of tumor cells as assessed through Cell Counting Kit-8, wounding healing assays, and Transwell assays. The upregulation of PLS3 also led to enhanced phosphatidylinositol-3 kinase/protein kinase B signaling in PACA cells. These data suggest that PLS3 is a biomarker to estimate PACA progression and represents a molecular target for PACA therapy.

Upregulation of DUSP14 Affects Proliferation, Invasion and Metastasis, Potentially via Epithelial-Mesenchymal Transition and Is Associated with Poor Prognosis in Pancreatic Cancer.

BackgroundThere is a growing number of evidence which report the relationship of the dual-specificity phosphatases 14 (DUSP14) with physiological and pathological mechanisms in the human body. However, it is still not known what if any role DUSP14 plays in pancreatic cancer.Materials and MethodsThe study evaluates the levels of DUSP14 in the pancreatic cancer tissues and cell lines using Western blotting and qRT-PCR to assess the levels of the DUSP14 and epithelial–mesenchymal transition (EMT) biomarkers. After the DUSP14 was blocked, the following assays were performed: colony formation, assessments of scratch wound and transwell to examine the effects of DUSP14 on the proliferation, migration and invasion of the pancreatic cancer.ResultsResults showed that there was a significant increase in the level of DUSP14 expression both in the pancreatic cancer tissues and cell lines. Experimental downregulation of DUSP14 induced the inhibition of the capacity of proliferation, migration and invasion of the pancreatic cancer cells. Western blotting analyses showed changes in the levels of expression of the EMT biomarkers, which helped to determine the function of DUSP14 in EMT.ConclusionIn conclusion, we suggest that DUSP14 is a novel molecular target that can be used for the treatment of pancreatic cancer.

Musashi2 promotes EGF-induced EMT in pancreatic cancer via ZEB1-ERK/MAPK signaling

BackgroundOur previous study showed Musashi2 (MSI2) promoted chemotherapy resistance and pernicious biology of pancreatic cancer (PC) by down-regulating Numb and p53. We further explored the novel molecular mechanism involving its oncogenic role in PC development.MethodsWe investigated the potential role and mechanism of MSI2 in EGF-induced EMT in PC in vitro and vivo.ResultsEGF enhanced EGFR (epidermal growth factor receptor) phosphorylation, induced EMT and activated ZEB1-ERK/MAPK signaling in 2 PC cells. However, MSI2 silencing reversed EGF stimulated function, including inhibiting EGF-promoted EMT-like cell morphology and EGF-enhanced cell invasion and migration. Meanwhile, MSI2 silencing inhibited EGF-enhanced EGFR phosphorylation at tyrosine 1068 and reversed EGF-induced change of the key proteins in EMT and ZEB1-ERK/MAPK signaling (ZEB1, E-cad, ZO-1, β-catenin, pERK and c-Myc). Additionally, MSI2 was co-stained and co-immunoprecipitated with ZEB1, pERK and c-Myc in PC cells by IF and co-IP, implying a close interaction between them. In vivo, MSI2 silencing inhibited pancreatic tumor size in situ and distant liver metastases. A close relationship of MSI2 with EMT and ZEB1-ERK/MAPK signaling were also observed in vivo and human PC samples, which coordinately promoted the poor prognosis of PC patients.ConclusionsMSI2 promotes EGF-induced EMT in PC via ZEB1-ERK/MAPK signaling.

High expression of olfactomedin-4 is correlated with chemoresistance and poor prognosis in pancreatic cancer.

Pancreatic cancer has an extremely poor prognosis, and identification of novel predictors of therapeutic efficacy and prognosis is urgently needed. Chemoresistance-related molecules are correlated with poor prognosis and may be effective targets for cancer treatment. Here, we aimed to identify novel molecules correlated with chemoresistance and poor prognosis in pancreatic cancer. We established 10 patient-derived xenograft (PDX) lines from patients with pancreatic cancer and performed next-generation sequencing (NGS) of tumor tissues from PDXs after treatment with standard drugs. We established a gene-transferred tumor cell line to express chemoresistance-related molecules and analyzed the chemoresistance of the established cell line against standard drugs. Finally, we performed immunohistochemical (IHC) analysis of chemoresistance-related molecules using 80 pancreatic cancer tissues. From NGS analysis, we identified olfactomedin-4 (OLFM4) as having high expression in the PDX group treated with anticancer drugs. In IHC analysis, OLFM4 expression was also high in PDXs administered anticancer drugs compared with that in untreated PDXs. Chemoresistance was observed by in vitro analysis of tumor cell lines with forced expression of OLFM4. In an assessment of tissue specimens from 80 patients with pancreatic cancer, Kaplan-Meier analysis showed that patients in the low OLFM4 expression group had a better survival rate than patients in the high OLFM4 expression group. Additionally, multivariate analysis showed that high expression of OLFM4 was an independent prognostic factor predicting poor outcomes. Overall, our study revealed that high expression of OLFM4 was involved in chemoresistance and was an independent prognostic factor in pancreatic cancer. OLFM4 may be a candidate therapeutic target in pancreatic cancer.

Lost miR-141 and upregulated TM4SF1 expressions associate with poor prognosis of pancreatic cancer: regulation of EMT and angiogenesis by miR-141 and TM4SF1 via AKT

ABSTRACT Background Transmembrane-4-L-six-family-1 (TM4SF1) functions to regulate cell growth and mobility and TM4SF1 expression was upregulated in pancreatic cancer. This study further investigated the role of TM4SF1 in regulating pancreatic cancer epithelial-mesenchymal transition (EMT) and angiogenesis and the underlying molecular events. Methods Tissue specimens were collected from 90 pancreatic cancer patients for immunohistochemical and qRT-PCR analysis of miR-141 and TM4SF1 levels, respectively. Pancreatic cancer cell lines were used for in vitro assays, while nude mice were used for the in vivo assay. Results TM4SF1 expression was upregulated, whereas miR-141 expression was lost in pancreatic cancer tissues, both of which was associated with advanced clinicopathological features and poor survival of pancreatic cancer patients. Furthermore, miR-141 was able to target and reduce TM4SF1 expression in pancreatic cancer cells and miR-141 expression inhibited pancreatic cancer cell EMT in vitro and Matrigel plug angiogenesis and lung metastasis in nude mice. At the gene level, miR-141 directly targeted and reduced TM4SF1 expression and in turn induced E-cadherin expression and reduced VEGF-A expression by suppressing activation of the AKT signaling pathway. Conclusions This study demonstrated that upregulated TM4SF1 and lost miR-141 expression were associated with advanced clinicopathological features and poor survival of pancreatic cancer patients. Lost miR-141 expression but induced TM4SF1 expression altered expression of VEGF-A and E-cadherin and promoted pancreatic cancer cell EMT and angiogenesis via the AKT signaling pathway, suggesting that targeting of miR-141 and TM4SF1 may be a potential therapeutic strategy to control pancreatic cancer.

LINC00473 functions as an oncogene and predicts poor prognosis in pancreatic cancer via the cAMP/β-catenin axis.

To investigate the expression and function of LINC00463 in pancreatic cancer (PC), and to demonstrate the relationship between LINC00473 expression and clinical pathological characteristics and prognosis of PC. Expressions of LINC00473 in PC tissues and cell lines were detected using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). LINC00473 siRNA was synthesized to knock down the LINC00473 expression in PANC-1 cells. Proliferation, invasion, and migration abilities of experimental cells were analyzed using cell counting kit-8 (CCK-8) assay and transwell assay, respectively. cAMP activity was detected and protein expression of β-catenin was measured to explain the underlying mechanism of LINC00473 in PC. The prognosis and clinical pathological features of PC patients were illustrated. LINC00473 was highly expressed in PC tissues and cells. Higher level of LINC00473 was relative with larger tumor size, worse tumor node metastasis (TNM) stage, worse tumor differentiation, higher rates of perineural invasion, and lymphatic invasion. Knockdown of LINC00473 significantly inhibited cell growth, invasion, and migration of PANC-1 cells. LINC00473 activated cAMP and then promoted the phosphorylation of β-catenin to promote the progression of PC. Furthermore, high expression of LINC00473 and β-catenin remarkedly indicated poor prognosis of PC patients. LINC00473 was upregulated in PC tissues and cells, indicating a poor prognosis and clinical pathological features of PC. It promoted PC progression via activating the cAMP/β-catenin axis, which provided a novel target for the prediction for PC diagnosis, biological therapy, and prognosis.

Prediction of Unresectability and Prognosis in Patients Undergoing Surgery on Suspicion of Pancreatic Cancer Using Carbohydrate Antigen 19-9, Interleukin 6, and YKL-40

The aim was to determine whether serum levels of carbohydrate antigen (CA) 19-9, interleukin 6 (IL-6), and YKL-40 could identify advanced disease and poor prognosis in pancreatic cancer (PC) patients intraoperatively diagnosed with locally advanced or metastatic disease. Two hundred ninety patients were included with preoperative blood samples. Plasma IL-6 and YKL-40 were determined by enzyme-linked immunosorbent assays. Interleukin 6 was elevated in patients with unresectable PC compared with resectable PC (P = 0.03). Carbohydrate antigen 19-9 and YKL-40 were similar. Patients with resectable tumors and greater than median preoperative CA 19-9, IL-6, and YKL-40 had shorter overall survival than patients with low levels (CA 19-9: hazard ratio [HR], 1.79; 95% confidence interval [CI], 1.13-2.83; P = 0.01; IL-6: HR, 1.83; 95% CI, 1.20-2.78; P = 0.01; YKL-40: HR, 1.60; 95% CI, 1.02-2.49; P = 0.04). Patients with resectable tumors and 2 or 3 high biomarker levels had significantly reduced overall survival compared with patients with low levels (2 high: HR, 2.97; 95% CI, 1.44-6.10; P = 0.00; 3 high: HR, 3.10; 95% CI, 1.45-6.65; P = 0.00). Preoperative levels of CA 19-9, IL-6, and YKL-40 may be useful to identify a subgroup of PC patients with poor prognosis.

LAMA4 upregulation is associated with high liver metastasis potential and poor survival outcome of Pancreatic Cancer.

Rationale: Pancreatic cancer is one of the most difficult cancers to manage and its poor prognosis stems from the lack of a reliable early disease biomarker coupled with its highly metastatic potential. Liver metastasis accounts for the high mortality rate in pancreatic cancer. Therefore, a better understanding of the mechanism(s) underlying the acquisition of the metastatic potential in pancreatic cancer is highly desirable.Methods: Microarray analysis in wild-type and highly liver metastatic human pancreatic cancer cell lines was performed to identify gene expression signatures that underlie the metastatic process. We validated our findings in patient samples, nude mice, cell lines and database analysis.Results: We identified a metastasis-related gene, laminin subunit alpha 4 (LAMA4), that was upregulated in highly liver metastatic human pancreatic cancer cell lines. Downregulation of LAMA4 reduced the liver metastatic ability of pancreatic cancer cells in vivo. Furthermore, LAMA4 expression was positively correlated with tumor severity and in silico analyses revealed that LAMA4 was associated with altered tumor microenvironment. In particular, our in vitro and in vivo results showed that LAMA4 expression was highly correlated with cancer-associated fibroblasts (CAFs) level which may contribute to pancreatic cancer metastasis. We further found that LAMA4 had a positive effect on the recruitment and activity of CAFs.Conclusions: These data provide evidence for LAMA4 as a possible biomarker of disease progression and poor prognosis in pancreatic cancer. Our findings indicate that LAMA4 may contribute to pancreatic cancer metastasis via recruitment or activation of CAFs.

HOXB5 promotes malignant progression in pancreatic cancer via the miR-6732 pathway

ABSTRACT Background: Homeobox B5 (HOXB5) is associated with the poor prognosis of various cancer types. However, the specific mechanism by which HOXB5 promotes the malignant progression of pancreatic cancer (PC) remains to be determined. Methods: The Cancer Genome Atlas database indicated HOXB5 expression level correlated to PC prognosis. The biological functions of HOXB5 was confirmed by colony formation, migration, and invasion assays. The effects of HOXB5 on the expression of cancer stem cell and epithelial–mesenchymal transition markers were evaluated. The downstream target of HOXB5 was miR-6723, which was detected by transcriptional assay. A xenograft tumor model was established in nude mice for the assessment of the role of HOXB5 in tumor growth and metastasis. Results: PC tissues had higher HOXB5 expression levels than noncancerous tissues, and high HOXB5 expression was significantly associated with poor PC prognosis. HOXB5 knockdown suppressed clone formation and the proliferation, invasion, and migration of PC cells in vitro. Conversely, these activities were enhanced by HOXB5 overexpression. The HOXB5 that bound two synergy motifs regulated miR-6723 expression and contributed to PC malignant progression. The role of HOXB5 in promoting tumor growth and metastasis was verified in vivo. Further investigation revealed that Twist1 and Zeb1 expression levels were increased by HOXB5. Conclusions: HOXB5 overexpression was significantly correlated with poor PC prognosis. HOXB5 accelerated the malignant progression of PC by up-regulating miR-6723, which afforded PC cells stem-like properties and facilitated the epithelial–mesenchymal transition of PC cells.

Abstract B52: The T-cell architecture of pancreatic ductal adenocarcinoma

Abstract Pancreatic cancer has the worst prognosis of any human malignancy. Our recent work has shown that immune infiltrate by transcriptomics and histopathology can predict prognosis after a Whipple’s operation, with lymphocyte infiltration being the most important prognostic marker. We have profiled T cells within primary pancreatic cancer to understand the T-cell architecture within the tumor. We have used a 37-marker T cell-focused panel to study the immune infiltrate. We see a diverse immunosuppressive immune microenvironment within the primary tumor. There is a complex architecture of macrophages, neutrophils, and T cells. The commonest CD4 T cells in the microenvironment are Tregs. There are potential novel therapeutic strategies for Tregs using unique checkpoint inhibitors. The CD8 T cells have PD-1 activity but are not activated or proliferating. The poor prognosis of pancreatic cancer could be explained by the immunosuppressive microenvironment, but there are opportunities to drug Tregs in this disease. Citation Format: Shivan Sivakumar, Enas Abu Shah, David Ahern, Nagina Mangal, Srikanth Reddy, Aniko Rendek, Zahir Soonawalla, Michael Silva, Mark Middleton, Michael Dustin. The T-cell architecture of pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2019 Sept 6-9; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2019;79(24 Suppl):Abstract nr B52.

Identification of Matrix Metalloproteinase 11 as a Prognostic Biomarker in Pancreatic Cancer.

The aim of this study was to investigate matrix metalloproteinase 11 (MMP11) as a promising biomarker in human pancreatic cancer. A consecutive eliminating method was used to select biomarker candidates in pancreatic cancer. mRNA and protein expression levels of candidates were determined in tissues and whole blood samples of healthy donors and pancreatic cancer patients. The prognostic value of MMP11 was determined using various data-sets and Liptak's Z analysis. Analysis using Gene Expression Omnibus (GEO) database showed significantly higher MMP11 mRNA expression in pancreatic cancer tissues compared to that in various normal tissues. MMP11 protein was specifically expressed in pancreatic cancer tissues, but not in various normal or other cancer tissues. Secreted MMP11 levels could be measured using easily accessible techniques and whole blood samples of pancreatic cancer. In addition, high levels of MMP11 were associated with poor prognosis of pancreatic cancer patients. MMP11 may be a promising prognostic biomarker for pancreatic cancer patients.

PCN470 EXPERT CONSENSUS ON PATIENT-REPORTED OUTCOMES FOR THE MANAGEMENT OF PANCREATIC CANCER

Systematic collection of patient-reported outcomes (PROs) is of increasing importance in cancer care since it improves symptom control and patient-physician communication. Given the poor prognosis of pancreatic cancer, accurate monitoring of symptoms becomes critical to improve patient’s outcomes, health status and care. This project aims to reach consensus on most significant PROs in pancreatic cancer, from both patients and healthcare professionals (HCP) perspectives, to be collected in clinical practice. The project was led by a Scientific Committee (SC) composed by 5 experts on pancreatic cancer management (n=3 oncologists, n=1 hospital pharmacist, n=1 nurse). A literature review (LR) was performed to identify PROs in pancreatic cancer. Based on the results of the LR and their clinical expertise, SC elaborated a list with PROs that was later presented to HCP and patients. Two nominal groups with HCP (n=10 oncologists, n=4 hospital pharmacists, n=2 nurses) and 6 semi-structured interviews with patients were conducted to establish the most valuable PROs (consensus ≥75% participants agree). SC determined those PROs that not achieved consensus among nominal groups and semi-structured interviews. A total of 49 disease-related signs and symptoms and/or its treatment that could be self-reported by the patient were identified during the LR (n=45) and by the SC (n=4). During nominal groups and semi-structured interviews 16 PROs were identified as the most significant. Of them, 6 reached consensus in both nominal groups and 9 were also considered valuable by patients. Finally, the SC selected 14 PROs: fever (≥38ºC), gastrointestinal bleeding, pain, edema, weight/appetite loss, diarrhea/constipation, nausea/vomiting, asthenia/fatigue, dyspnea, emotional wellbeing, neuropathy, ascites, change in the color of urine/stools and/or skin/eyes, swallowing problems. The findings of the present project are the first step to implement the systematic collection of PROs that may enhance patient care and patient-physician communication in clinical practice.

ERO1α promotes hypoxic tumor progression and is associated with poor prognosis in pancreatic cancer.

Pancreatic cancer is a leading cause of mortality worldwide due to the difficulty of detecting early-stage disease and our poor understanding of the mediators that drive progression of hypoxic solid tumors. We therefore used a heavy isotope ‘pulse/trace’ proteomic approach to determine how hypoxia (Hx) alters pancreatic tumor expression of proteins that confer treatment resistance, promote metastasis, and suppress host immunity. Using this method, we identified that hypoxia stress stimulates pancreatic cancer cells to rapidly translate proteins that enhance metastasis (NOTCH2, NCS1, CD151, NUSAP1), treatment resistance (ABCB6), immune suppression (NFIL3, WDR4), angiogenesis (ANGPT4, ERO1α, FOS), alter cell metabolic activity (HK2, ENO2), and mediate growth-promoting cytokine responses (CLK3, ANGPTL4). Database mining confirmed that elevated gene expression of these hypoxia-induced mediators is significantly associated with poor patient survival in various stages of pancreatic cancer. Among these proteins, the oxidoreductase enzyme ERO1α was highly sensitive to induction by hypoxia stress across a range of different pancreatic cancer cell lines and was associated with particularly poor prognosis in human patients. Consistent with these data, genetic deletion of ERO1α substantially reduced growth rates and colony formation by pancreatic cancer cells when assessed in a series of functional assays in vitro. Accordingly, when transferred into a mouse xenograft model, ERO1α-deficient tumor cells exhibited severe growth restriction and negligible disease progression in vivo. Together, these data indicate that ERO1α is potential prognostic biomarker and novel drug target for pancreatic cancer therapy.