Pancreatic cancer is an aggressive cancer of the digestive system, which is becoming a serious health problem worldwide. Overall survival for patients with pancreatic cancer is poor, mainly due to a lack of biomarkers to enable early diagnosis and a lack of prognostic markers that can inform decision-making, facilitating personalized treatment and an optimal clinical outcome. ncRNAs play an important role in pancreatic carcinogenesis. Here we review the literature on the role of ncRNAs as biomarkers in pancreatic cancer. We focus on the significance of ncRNAs as markers for early diagnosis, as prognostic biomarkers able to inform clinical management and as targets for novel therapeutics for patients with pancreatic cancer.

Although biliary tract cancers (BTCs) are known to have an inflammatory component, a detailed characterisation of immune-related transcripts has never been performed. In these studies, nCounter PanCancer Immune Profiling Panel was used to assess the expression of 770 immune-related transcripts in the tumour tissues (TTs) and matched adjacent tissues (ATs) of resected BTCs. Cox regression analysis and Kaplan–Meier methods were used to correlate findings with relapse-free survival (RFS). The first analysis in the TT and AT of an exploratory set (n = 22) showed deregulation of 39 transcripts associated with T-cell activation. Risk of recurrence was associated with a greater number of genes deregulated in AT in comparison to TT. Analysis in the whole set (n = 53) showed a correlation between AT cytotoxic T-lymphocyte antigen-4 (CTLA4) expression and RFS, which maintained statistical significance at multivariate analysis. CTLA4 expression correlated with forkhead box P3 (FOXP3) expression, suggesting enrichment in T regulatory cells. CTLA4 is known to act by binding to the cluster of differentiation 80 (CD80). No association was seen between AT CD80 expression and RFS. However, CD80 expression differentiated prognosis in patients who received adjuvant chemotherapy. We showed that the immunomodulatory transcriptome is deregulated in resected BTCs. Our study includes a small number of patients and does not enable to draw definitive conclusions; however, it provides useful insights into potential transcripts that may deserve further investigation in larger cohorts of patients.Transcript ProfilingNanostring data have been submitted to GEO repository: GSE90698 and GSE90699.

FOLFIRINOX, a combination of chemotherapy drugs (Fluorouracil, Oxaliplatin, Irinotecan -FOI), provides the best clinical benefit in pancreatic ductal adenocarcinoma (PDAC) patients. In this study we explore the role of miRNAs (MIR) as modulators of chemosensitivity to identify potential biomarkers of response. We find that 41 and 84 microRNA inhibitors enhance the sensitivity of Capan1 and MiaPaCa2 PDAC cells respectively. These include a MIR1307-inhibitor that we validate in further PDAC cell lines. Chemotherapy-induced apoptosis and DNA damage accumulation are higher in MIR1307 knock-out (MIR1307KO) versus control PDAC cells, while re-expression of MIR1307 in MIR1307KO cells rescues these effects. We identify binding of MIR1307 to CLIC5 mRNA through covalent ligation of endogenous Argonaute-bound RNAs cross-linking immunoprecipitation assay. We validate these findings in an in vivo model with MIR1307 disruption. In a pilot cohort of PDAC patients undergoing FOLFIRONX chemotherapy, circulating MIR1307 correlates with clinical outcome.

ntroduction: FOLFIRINOX-regime is a combination-chemotherapy that provides the best clinical benefit in pancreatic cancer (PC) patients, but is associated with severe toxicity. Aim of this study is to explore the role of miRNAs (MIR) as modulators of chemosensitivity and their potential as biomarkers of sensitivity to FOLFIRINOX-chemotherapy. Methods: High-throughput-screening (HTS) of 997 LNA-MIR-inhibitors was performed in PC cell lines (Capan1, MiaPaCa2) treated with a combination of Fluorouracil (F), Oxaliplatin (O) and Irinotecan (I) that mimics FOLFIRINOX-regime. Cell viability was monitored by CellTiter-Blue assay. Validation experiments were carried out with miRvana probes. MIR expression was assessed by TaqMan-assay. Apoptosis was measured by Flow-cytometry and western-blotting. Knock-out of microRNA was acheived by CRISPR-CAS9 in MiaPaca2 cells (MIR1307KO). Results: 41 and 84 miRNA-inhibitors enhanced FOI activity by >30% (p<0.001) in Capan1 and MiaPaCa2. These included MIR1307-inhibitor that was validated in Capan1, MiaPaCa2, Panc1, AspC1, BxPC3, and Su86.86 cell lines. The proportion of cells killed by FOI in comparison to DMSO was greater in cells transfected with MIR1307 inhibitor when compared to NEG CTRL, making the effect of this MIR specific for chemotherapy. MIR1307 was over-expressed in tumour compared to matched-adjacent tissue in 40/60 human PC cases, confirming clinical relevance. MIR1307KO cells were more sensitive to FOI than WT cells. Chemotherapy-induced apoptosis was higher in MIR1307KO cells (caspase 3/7 activity and annexin-V positivity). We observed significant upregulation of different markers of DNA damage (pH2AX2, 8OHdG, DNA breaks in Comet assay) in MIR1307KO cells treated with FOI in comparison to WT treated cells. Re-expression of MIR1307 in MIR1307KO cells could increase resistance to FOI chemotherapy and protected from FOI-induced DNA damage. Bioinformatics analysis identified MIR1307 binding-sites within a number of genes involved in the DNA-repair pathway (p<0.001, folding energy value greater than -12 Kcal/mol). Conclusions: We identified miR-1307 as a potential modulator of sensitivity to FOI-chemotherapy in PC. We showed that miR-1307 inhibition impairs the ability of PC cells to recover from chemotherapy damage and therefore enhances its activity. The assessemnt of it potential as predictive biomarker of response in PC patients is ongoing. Citation Format: Pietro Carotenuto, Domenico Zito, Maria C. Previdi, Maya Raj, Matteo Fassan, Andrea Lampis, Francesco Scalafani, Andrea Lanese, Ian Said-Huntingford, Jens C. Hahne, Kate Young, Ruwaida Begum, Zakaria Ethiar, Andrew Wotherspoon, Naureen Starling, Anguraj Sadanandam, David Cunningham, Ian Chau, Paul Workman, Rajesh Chopra, Nicola Valeri, Chiara Braconi. MIR1307 mediates pancreatic cancer resistance to FOLFIRINOX chemotherapy by affecting response to DNA damage [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4977.

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