Search

This deliverable D3.5 is an update on training activities performed within task 3.1 in the second period of the VISION project implementation. Short-term academic stays of early-stage researchers in partners' laboratories allowed the applicants to improve and acquire new scientific and technical skills in various cellular, molecular, and genetic techniques involved in basic and clinical research. Moreover, they had the opportunity to strengthen their personal and professional development in gastrointestinal cancer and nanobiotechnology. The involvement of early-stage researchers in such scientific events enhanced the reputation, credibility, attractiveness, and networking activities of BMC SAV at both international and national levels. Besides BMC SAV, also other VISION partners sent their early-stage scientists for short-term academic stays to partner laboratories. The coronavirus pandemic delayed the performance of training activities in 2020. In order to overcome these objective limitations in implementing the planned actions, and if it was possible, the training activities were replaced by theoretical courses and e-lectures (D3.1 Training activities). After the travel restrictions were removed, the first short-term early-stage researchers exchanges started in 2021, and the last academic stay finished in June 2023. At the 2nd Partner Assembly meeting in September 2021, the partners agreed to fully open all in-person training activities since January 2022. In total (2020-2023), 22 in-person training activities have been realized, and 28 early-stage researchers have participated. Some applicants were interested in more than one training.

Several joint research publications were generated and submitted to peer-reviewed journals during the VISION project as a result of close collaboration among the consortium partners. In each publication, the authors acknowledged the VISION project, which has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 857381. In all cases, the consortium opted for golden open-access journals, or those free of charge with online access for any user. In total, 26 publications have been published and several others are under revision or evaluation. All publications are available at the VISION website - http://vision.sav.sk/publications.html The open access to these publications, as well as other project outputs, is also granted in the ZENODO repository, in the community “VISION H2020 Project” - https://www.zenodo.org/communities/vision-h2020/?page=1&size=20.

Tumor organoids are three-dimensional (3D) ex vivo tumor models that recapitulatethe biological key features of the original primary tumor tissues. Patient-derivedtumor organoids have been used in translational cancer research and can beapplied to assess treatment sensitivity and resistance, cell-cell interactions, and tumorcell interactions with the tumor microenvironment. Tumor organoids are complexculture systems that require advanced cell culture techniques and culture mediawith specific growth factor cocktails and a biological basement membrane thatmimics the extracellular environment. The ability to establish primary tumor cultureshighly depends on the tissue of origin, the cellularity, and the clinical featuresof the tumor, such as the tumor grade. Furthermore, tissue sample collection,material quality and quantity, as well as correct biobanking and storage are crucialelements of this procedure. The technical capabilities of the laboratory are also crucialfactors to consider. Here, we report a validated SOP/protocol that is technically andeconomically feasible for the culture of ex vivo tumor organoids from fresh tissuesamples of pancreatic adenocarcinoma origin, either from fresh primary resectedpatient donor tissue or patient-derived xenografts (PDX). The technique describedherein can be performed in laboratories with basic tissue culture and mouse facilitiesand is tailored for wide application in the translational oncology field.

Cancer is a complex and heterogeneous disease that arises from the accumulation of geneticand epigenetic alterations in cells. Tumor progression and metastasis are the major contributorsto cancer-related deaths worldwide. Understanding their underlying genetic and epigeneticmechanisms is essential for the development of effective diagnostic and therapeutic approaches. The aim of this reprint is to provide a wide range of topics, including the cellular and molecularmechanisms underlying tumor progression and metastasis, involvement of cell signaling pathways,cellular plasticity, and interactions with the tumor microenvironment. This reprint also discussesthe role of oncogenes and tumor suppressor genes, chromosomal rearrangements, copy numbervariations, and epigenetic modifications. Furthermore, it provides the current state of the art inseveral aspects of cancer diagnosis, prevention, and treatment and highlights emerging therapeuticstrategies based on genetic and epigenetic alterations. The contributing authors are renowned experts in the field of cancer research and treatment.They provide valuable insights into the latest discoveries and advances in cancer research and offer aunique perspective on the challenges and opportunities in the fight against cancer. We hope this reprint will serve as a valuable resource for researchers, clinicians, and studentsworking in the field of cancer biology and oncology. We believe that the knowledge and insightspresented here will inspire further research and innovation in the field of cancer research.

Deliverable D4.2 describes the tools that have been utilized to widen learning opportunities for undergraduate students and early-stage researchers beyond a formal education system. The main objective of Open school was to allow targeted groups time and location-free access to up-to-date knowledge in the field of molecular biology, genetics, liquid biopsy in oncology, immuno-oncology, nanobiology, and nanomedicine.

Genetic aberrations, including chromosomal rearrangements, loss or amplification ofDNA, and point mutations, are major elements of cancer development. However, sinceepigenetic dysregulation was shown to be strongly related to human disease, especiallycancer, the epigenetic component seems equally important. The term epigenetics wasfirst introduced by ConradWaddington in 1942 [1] in his effort to link the genotype withphenotype. It is currently used to describe the ensemble of several mechanisms thatcan reversibly modify gene expression profiles without altering the DNA sequence [2].Epigenetic mechanisms are meant to cooperate with various other regulatory factors tosecure time and tissue-specific regulation of gene expression in relation to developmentalor environmental cues.

Pancreatic neuroendocrine neoplasms (pNENs) are rare cancers with broad challenges for their management. Themain clinical obstacles are the high rate of patients diagnosed at advanced stages, lack of prognostic markers forearly detection of disease recurrence in resected patients, significant limitations in identifying those who willbenefit from adjuvant therapy, and timely recognition of treatment response. Therefore, the discovery of newprognostic and predictive markers is necessary for patient stratification and clinical management. Liquid biopsy,which has revolutionized the field of clinical oncology, is extremely under-investigated in pNENs. This reviewhighlights its potential and the recent advances in related technologies, as candidates for the delivery of the newtools that can help to refine pNEN diagnosis and to personalize treatment. In addition, the opportunities andlimitations of available preclinical research models with regard to biomarker research are discussed in light ofpNEN clinical needs.

Training courses were organised by the coordinator in collaboration with the consortium partners NILU, FhG, NKUA and SERMAS for early stage researchers to acquire new skills and knowledge in the fields of scientific writing, interacting with industry, proposal writing and project management as well as to receive more theoretical background in specific technical methods. In total, 13 different courses in the fields mentioned above were realized as online events until now. Courses 1-4 took place in 2020 and 2021, so in total 17 courses were organized until September 2022. Two more courses are planned until the end of 2022. The training courses were organized as open call events for all students on the VISION website, not limited for students at BMC SAV in Slovakia.

The term liquid biopsy (LB) refers to molecules such as proteins, DNA, RNA, cells, or extracellular vesicles in blood and other bodily fluids that originate from the primary and/or metastatic tumor. LB has emerged as a mainstay in translational research and has started to become part of clinical oncology practice, providing a minimally invasive alternative to solid biopsy. The LB allows real-time monitoring of a tumor via a minimally invasive sample extraction, such as blood. The applications include early cancer detection, patient follow-up for the detection of disease progression, assessment of minimal residual disease, and potential identification of molecular progression and mechanism of resistance. In order to achieve a reliable analysis of these samples that can be reported in the clinic, the preanalytical procedures should be carefully considered and strictly followed. Sample collection, quality, and storage are crucial steps that determine their usefulness in downstream applications. Here, we present standardized protocols from our liquid biopsy working module for collecting, processing, and storing plasma and serum samples for downstream liquid biopsy analysis based on circulating-free DNA. The protocols presented here require standard equipment and are sufficiently flexible to be applied in most laboratories focused on biological procedures.

Gastrointestinal (GI) cancers are malignancies that develop within the digestive system and account for one in four cancer cases according to WHO data [...]

Due to abundant stroma and extracellular matrix, accompanied by lack of vascularization,pancreatic ductal adenocarcinoma (PDAC) is characterized by severe hypoxia. Epigenetic regulationis likely one of the mechanisms driving hypoxia-induced epithelial-to-mesenchymal transition (EMT),responsible for PDAC aggressiveness and dismal prognosis. To verify the role of DNA methylationin this process, we assessed gene expression and DNA methylation changes in four PDAC cell lines.BxPC-3, MIA PaCa-2, PANC-1, and SU.86.86 cells were exposed to conditioned media containingcytokines and inflammatory molecules in normoxic and hypoxic (1% O2) conditions for 2 and 6 days.Cancer Inflammation and Immunity Crosstalk and Human Epithelial to Mesenchymal TransitionRT2 Profiler PCR Arrays were used to identify top deregulated inflammatory and EMT-related genes.Their mRNA expression and DNA methylation were quantified by qRT-PCR and pyrosequencing.BxPC-3 and SU.86.86 cell lines were the most sensitive to hypoxia and inflammation. Although themethylation of gene promoters correlated with gene expression negatively, it was not significantlyinfluenced by experimental conditions. However, DNA methyltransferase inhibitor decitabineefficiently decreased DNA methylation up to 53% and reactivated all silenced genes. These resultsconfirm the role of DNA methylation in EMT-related gene regulation and uncover possible newtargets involved in PDAC progression.

In recent years, it has been verified that collective cell migration is a fundamental step in tumor spreading and metastatic processes. In this paper, we demonstrate for the first time how low-intensity ultrasound produces long-term inhibition of collective migration of epithelial cancer cells in wound healing processes. In particular, we show how pancreatic tumor cells, PANC-1, grown as monolayers in vitro respond to these waves at frequencies close to 1 MHz and low intensities (−2) for 48-72 hours of culture after ultrasound irradiation. This new strategy opens a new line of action to block the spread of malignant cells in cancer processes. Despite relevant spatial variations of the acoustic pressure amplitude induced in the assay, the cells behave as a whole, showing a collective dynamic response to acoustic performance, agreeing with the “cell jamming” effect defined before in the literature. Experiments carried out with samples without previous starving showed remarkable effects of the LICUs from the first hours of culture, more prominent than those with experiments with monolayers subjected to fasting prior to the experiments. This new strategy to control cell migration opens a new line of action in vivo to block the spread of malignant cells.

INTRODUCTION: The IMMray PanCan-d test combines an 8-plex biomarker signature with CA19-9 in a proprietary algorithm to detect pancreatic ductal adenocarcinoma (PDAC) in serum samples. This study aimed to validate the clinical performance of the IMMray PanCan-d test and to better understand test performance in Lewis-null (le/le) individuals who cannot express CA19-9. METHODS: Serum samples from 586 individuals were analyzed with the IMMray PanCan-d biomarker signature and CA19-9 assay, including 167 PDAC samples, 203 individuals at high risk of familial/hereditary PDAC, and 216 healthy controls. Samples were collected at 11 sites in the United States and Europe. The study was performed by Immunovia, Inc (Marlborough, MA), and sample identity was blinded throughout the study. Test results were automatically generated using validated custom software with a locked algorithm and predefined decision value cutoffs for sample classification. RESULTS: The IMMray PanCan-d test distinguished PDAC stages I and II (n = 56) vs high-risk individuals with 98% specificity and 85% sensitivity and distinguished PDAC stages I-IV vs high-risk individuals with 98% specificity and 87% sensitivity. We identified samples with a CA19-9 value of 2.5 U/mL or less as probable Lewis-null (le/le) individuals. Excluding these 55 samples from the analysis increased the IMMray PanCan-d test sensitivity to 92% for PDAC stages I-IV (n = 157) vs controls (n = 379) while maintaining specificity at 99%; test sensitivity for PDAC stages I and II increased from 85% to 89%. DISCUSSION: These results demonstrate the IMMray PanCan-d blood test can detect PDAC with high specificity (99%) and sensitivity (92%).

[Objective]: The lysyl oxidase-like protein 2 (LOXL2) contributes to tumour progression and metastasis in different tumour entities, but its role in pancreatic ductal adenocarcinoma (PDAC) has not been evaluated in immunocompetent in vivo PDAC models., [Design]: Towards this end, we used PDAC patient data sets, patient-derived xenograft in vivo and in vitro models, and four conditional genetically-engineered mouse models (GEMMS) to dissect the role of LOXL2 in PDAC. For GEMM-based studies, K-Ras +/LSL-G12D;Trp53 LSL-R172H;Pdx1-Cre mice (KPC) and the K-Ras +/LSL-G12D;Pdx1-Cre mice (KC) were crossed with Loxl2 allele floxed mice (Loxl2Exon2 fl/fl) or conditional Loxl2 overexpressing mice (R26Loxl2 KI/KI) to generate KPCL2KO or KCL2KO and KPCL2KI or KCL2KI mice, which were used to study overall survival; tumour incidence, burden and differentiation; metastases; epithelial to mesenchymal transition (EMT); stemness and extracellular collagen matrix (ECM) organisation., [Results]: Using these PDAC mouse models, we show that while Loxl2 ablation had little effect on primary tumour development and growth, its loss significantly decreased metastasis and increased overall survival. We attribute this effect to non-cell autonomous factors, primarily ECM remodelling. Loxl2 overexpression, on the other hand, promoted primary and metastatic tumour growth and decreased overall survival, which could be linked to increased EMT and stemness. We also identified tumour-associated macrophage-secreted oncostatin M (OSM) as an inducer of LOXL2 expression, and show that targeting macrophages in vivo affects Osm and Loxl2 expression and collagen fibre alignment., [Conclusion]: Taken together, our findings establish novel pathophysiological roles and functions for LOXL2 in PDAC, which could be potentially exploited to treat metastatic disease., JCL-G received support from a 'la Caixa' Foundation (ID 100010434) fellowship (LCF/BQ/DR21/11880011). This study was supported by ISCIII FIS grants PI18/00757 and PI21/01110 (BSJ) and PI18/00267 (LG-B), and grants from the Spanish Ministry of Economy and Innovation SAF2016-76504-R (ACan and FP), PID2019-111052RB-I00 (FP), PID2019-104644RB-I00 (GM-B), a Ramón y Cajal Merit Award RYC-2012–12104 (BSJ) and ISCIII, CIBERONC, CB16/12/00446 (ACar) and CB16/12/00295 (ACan and GM-B), all of them co-financed through Fondo Europeo de Desarrollo Regional (FEDER) 'Una manera de hacer Europa'; a Fero Foundation Grant (BSJ); a Coordinated grant (GC16173694BARB) from the Fundación Científica Asociación Española Contra el Cáncer (FC-AECC) (BSJ); a Miguel Servet award (CP16/00121) (PS); a DFG, German Research Foundation Grant—Project no: 492 436 553 (KG); and a Max Eder Fellowship of the German Cancer Aid (111746) (PCH)

Work package 4 is dedicated to mining and bridging the gaps between research and clinics. As cancer patients’ treatment is becoming more and more complex the concept of a multidisciplinary team working for patient’s benefit is considered the gold standard for improving cancer care. Within cancer multidisciplinary team surgeons, oncologists, radiologists, pathologists, specialist cancer nurses, physicians, specialists in basic scientists and psychologists work the best way to treat cancer patients. A successful multidisciplinary health team leads to a comprehensive assessment of a patient’s situation and works out a safer approach to treat each patient. Therefore, within the VISION project, it was foreseen that medical doctors during their specialization but also scientists will participate in such multidisciplinary teams within the hospital health care units participating in the project.

© 2021 by the authors., First-line treatment with regorafenib in frail metastatic colorectal cancer (mCRC) patients has shown some benefit. To accurately identify such patients before treatment, we studied blood biomarkers and primary tumor molecules. We unveiled serum microRNAs (miRNAs), single-nucleotide polymorphisms (SNPs) in angiogenic-related genes, and Notch 1 expression as biomarkers associated with response or toxicity. MicroRNA array profiling and genotyping of selected SNPs were performed in the blood of fragile mCRC patients treated with regorafenib. Notch 1 and CRC-associated miRNA expression was also analyzed in tumors. High levels of miR-185-5p in serum, rs7993418 in the vascular endothelial growth factor receptor 1 (VEGFR1) gene, and Notch 1 expression in biopsies were associated with a favorable response to treatment. Serum levels of miR-126-3p and miR-152-3p and tumor expression of miR-92a-1-5p were associated with treatment toxicity, particularly interesting in patients exhibiting comorbidities, and high levels of miR-362-3p were associated with asthenia. Additionally, several miRNAs were associated with the presence of metastasis, local recurrence, and peritoneal metastasis. Besides, miRNAs determined in primary tumors were associated with tumor-node-metastasis (TNM) staging. The rs2305948 and rs699947 SNPs in VEGFR2 and VEGFA, respectively, were markers of poor prognosis correlating with locoregional relapse, a higher N stage, and metastatic shedding. In conclusion, VEGF and VEGFR SNPs, miRNAs, and Notch 1 levels are potential useful biomarkers for the management of advanced CRC under regorafenib treatment., Financial support for this research was provided by Bayer Hispania, which had no role in the design of the study; the collection, analysis, and interpretation of data; and the writing of the manuscript.

© 2021 by the authors., Pancreatic ductal adenocarcinoma (PDAC) presents many challenges in the clinic and there are many areas for improvement in diagnostics and patient management. The five-year survival rate is around 7.2% as the majority of patients present with advanced disease at diagnosis that is treatment resistant. Approximately 10–15% of PDAC cases have a hereditary basis or Familial Pancreatic Cancer (FPC). Here we demonstrate the use of circulating free DNA (cfDNA) in plasma as a prognostic biomarker in PDAC. The levels of cfDNA correlated with disease status, disease stage, and overall survival. Furthermore, we show for the first time via BEAMing that the majority of hereditary or familial PDAC cases (around 84%) are negative for a KRAS somatic mutation. In addition, KRAS mutation negative cases harbor somatic mutations in potentially druggable genes such as KIT, PDGFR, MET, BRAF, and PIK3CA that could be exploited in the clinic. Finally, familial or hereditary cases have a longer overall survival compared to sporadic cases (10.2 vs. 21.7 months, respectively). Currently, all patients are treated the same in the clinic with cytotoxic agents, although here we demonstrate that there are different subtypes of tumors at the genetic level that could pave the way to personalized treatment., This study was funded by the Instituto de Salud Carlos III (Plan Estatal de I+D+i 2013– 2016): ISCIII (PI09/02221, PI12/01635, PI15/02101, and PI18/0135) and co-financed by the European Development Regional Fund “A way to achieve Europe” (ERDF), the Biomedical Research Network in Cancer: CIBERONC (CB16/12/00446), Red Temática de investigación cooperativa en cáncer: RTICC (RD12/0036/0073), La Asociación Española contra el Cáncer: AECC (Grupos Coordinados Estables 2016), Fundación Mutua Madrileña (FMM) / XVI Convocatoria de Ayudas a la Investigación en Salud 2019 and Asociación Cáncer de Páncreas (ACanPan); Asociación Española de Pancreatología (AESPANC) / IV Becas de Investigación Carmen Delgado/Miguel Pérez-Mateo 2019. The European Union’s Horizon 2020 research and innovation program under grant agreement No 857381, project VISION (Strategies to strengthen scientific excellence and innovation capacity for early diagnosis of gastrointestinal cancers).

Background: Early diagnosis is crucial for patients with pancreatic ductal adenocarcinoma (PDAC). The AXL receptor tyrosine kinase is proteolytically processed releasing a soluble form (sAXL) into the blood stream. Here we explore the use of sAXL as a biomarker for PDAC. Methods: AXL was analysed by immunohistochemistry in human pancreatic tissue samples. RNA expression analysis was performed using TCGA/GTEx databases. The plasma concentrations of sAXL, its ligand GAS6, and CA19-9 were studied in two independent cohorts, the HMar cohort (n = 59) and the HClinic cohort (n = 142), including healthy controls, chronic pancreatitis (CP) or PDAC patients, and in a familial PDAC cohort (n = 68). AXL expression and sAXL release were studied in PDAC cell lines and murine models. Findings: AXL is increased in PDAC and precursor lesions as compared to CP or controls. sAXL determined in plasma from two independent cohorts was significantly increased in the PDAC group as compared to healthy controls or CP patients. Patients with high levels of AXL have a lower overall survival. ROC analysis of the plasma levels of sAXL, GAS6, or CA19-9 in our cohorts revealed that sAXL outperformed CA19-9 for discriminating between CP and PDAC. Using both sAXL and CA19-9 increased the diagnostic value. These results were validated in murine models, showing increased sAXL specifically in animals developing PDAC but not those with precursor lesions or acinar tumours., This work was supported by grants from the Spanish Ministry of Science and Innovation (MICINN)/ Instituto de Salud Carlos III (ISCIII)-European Regional Development Fund (ERDF) (PI20/00625) and the “Generalitat de Catalunya” (2017/SGR/225) to PN; grants from the MICINN (Project# RTI2018-095672-BI00) to AM and PGdF; grant from the MICINN/ ISCIII-FEDER (PI20/01696) to MG and grant from the MICINN/ ISCIII-FEDER (PI18/01034) to AC. We also acknowledge grants from the Instituto de Salud Carlos III (ISCIII)/ERDF(PT20/00023) and the "Xarxa de Bancs de tumours” sponsored by Pla Director d'Oncologia de Catalunya (XBTC). PN’s lab is part of Redes de investigacion (Enfermedades Metabolicas y Cancer RED2018-102799-T), a project run by MINECO. CIBEREHD is funded by Instituto de Salud Carlos III.

Tumor cell dissemination is mediated by a subpopulation of cells referred to as circulating tumor cells (CTCs), which have epithelial to mesenchymal properties and an inherent capacity to intravasate from the primary tumor, extravasate into secondary organs, and survive both in circulation and within the secondary organ. These properties have long been assumed to be cell autonomous; however, it is now clear that CTCs require the aid of secondary cells, such as macrophages. The role of tumor-associated macrophages (TAMs) as direct and indirect players in tumor cell dissemination will be discussed in this chapter. Special emphasis will be placed on the interplay between CTC and macrophages, focusing on the pro-tumorigenic function of TAMs at the level of secreted proteins, how TAMs enhance the invasive nature of tumor cells, the role of TAMs in promoting inflammation and a pro-tumor immune response, and the important role TAMs play in metastasis. Macrophages as therapeutic targets will also be discussed. Finally, the concept that macrophages can physically interact with CTCs or fuse with tumor cells to create giant macrophages or cell fusions that are invisible to the immune system and have enhanced migratory and metastatic capacities will be presented and detailed.

8 páginas, 2 tablas, Ultrasounds (US) are a non-ionizing mechanical wave, with less adverse effects than conventional pharmacological or surgical treatments. Different biological effects are induced in tissues and cells by ultrasound actuation depending on acoustic parameters, such as the wave intensity, frequency and treatment dose. This non-ionizing radiation has considerable applications in biomedicine including surgery, medical imaging, physical therapy and cancer therapy. Depending on the wave intensity, US are applied as high-intensity ultrasounds (HIUS) and low-intensity pulsed ultrasounds (LIPUS), with different effects on cells and tissues. HIUS produce thermal and mechanical effects, resulting in a large localized temperature increase, leading to tissue ablation and even tumor necrosis. This can be achieved by focusing low intensity waves emitted from different electrically shifted transducers, known as high-intensity focused ultrasounds (HIFU). LIPUS have been used extensively as a therapeutic, surgical and diagnostic tool, with diverse biological effects observed in tissues and cultured cells. US represent a non-invasive treatment strategy that can be applied to selected areas of the body, with limited adverse effects. In fact, tumor ablation using HIFU has been used as a curative treatment in patients with an early-stage pancreatic tumor and is an effective palliative treatment in patients with advanced stage disease. However, the biological effects, dose standardization, benefit-risk ratio and safety are not fully understood. Thus, it is an emerging field that requires further research in order to reach its full potential., The authors would like to acknowledge the following funding related with the effects of ultrasounds on cancer cells and tissues: Spanish National Plan project RETOS DPI 2017-90147-R. Intramural call for new research projects for clinical researchers and emerging research groups. IRYCIS. (2018/0240). Ibero-American Network CYTED-DITECROD-218RT0545.

All consortium partners who take part in clinically orientated international and national meetings and conferences will promote theVISION project during oral and poster presentations. Either the Vision logo will be used in any presented work or the project will bereferenced in the acknowledgements.

Involvement of VISION partners in mentoring and co-supervision of PhD will increase the quality of education at universities, mainly medical and natural science faculties. PhD students of the Cancer Research Institute BMC SAS (Bratislava) are supervised by VISION project partners in the form of a co-supervision. So far seven PhD students have been allocated specific co-supervisors. Partner NILU supervises three PhD students, FhG and IRYCIS both each two PhD students. One of the IRYCIS co-supervised student will additionally be co-supervised by NKUA.

Training courses were organised by the coordinator in collaboration with the consortium partners NILU and FhG for early stageresearchers to acquire new skills and knowledge in the fields of scientific writing, interacting with industry, proposal writing andproject management. Four courses in the fields mentioned above were realized until now. The training courses were organized as open call events for all students, not limited for students at BMC SAV in Slovakia.

The 5-year survival rate of patients with pancreatic ductal adenocarcinoma (PDAC) is around 5% due to the fact that the majority of patients present with advanced disease that is treatment resistant. Familial pancreatic cancer (FPC) is a rare disorder that is defined as a family with at least two affected first degree relatives, with an estimated incidence of 4%-10%. The genetic basis is unknown in the majority of families although around 10%-13% of families carry germline mutations in known genes associated with hereditary cancer and pancreatitis syndromes. Panel sequencing was performed of 35 genes associated with hereditary cancer in 43 PDAC cases from families with an apparent hereditary pancreatic cancer syndrome. Pathogenic variants were identified in 19% (5/26) of PDAC cases from pure FPC families in the genes MLH1, CDKN2A, POLQ and FANCM. Low frequency potentially pathogenic VUS were also identified in 35% (9/26) of PDAC cases from FPC families in the genes FANCC, MLH1, PMS2, CFTR, APC and MUTYH. Furthermore, an important proportion of PDAC cases harboured more than one pathogenic, likely pathogenic or potentially pathogenic VUS, highlighting the multigene phenotype of FPC. The genetic basis of familial or hereditary pancreatic cancer can be explained in 21% of families by previously described hereditary cancer genes. Low frequency variants in other DNA repair genes are also present in 35% of families which may contribute to the risk of pancreatic cancer development. This study was funded by the Instituto de Salud Carlos III (Plan Estatal de I + D + i 2013-2016): ISCIII (PI09/02221, PI12/01635, PI15/02101 and PI18/1034) and co-financed by the European Development Regional Fund ''A way to achieve Europe'' (ERDF), the Biomedical Research Network in Cancer: CIBERONC (CB16/12/00446), Red Temática de investigación cooperativa en cáncer: RTICC (RD12/0036/0073) and La Asociación Española contra el Cáncer: AECC (Grupos Coordinados Estables 2016). This study was funded by the Instituto de Salud Carlos III (Plan Estatal de I+D+i 2013-2016): ISCIII (PI09/02221, PI12/01635, PI15/02101 and PI18/1034) and co-financed by the European Development Regional Fund "A way to achieve Europe'' (ERDF), the Biomedical Research Network in Cancer: CIBERONC (CB16/12/00446), Red Tematica de investigacion cooperativa en cancer: RTICC (RD12/0036/0073) and La Asociacion Espanola contra el Cancer: AECC (Grupos Coordinados Estables 2016). Sí

Pancreatic ductal adenocarcinoma (PDA) is the most frequent type of pancreatic cancer and one of the deadliest diseases overall. New biomarkers are urgently needed to allow early diagnosis, one of the only factors that currently improves prognosis. Here we analyzed whether the detection of circulating galectin-1 (Gal-1), a soluble carbohydrate-binding protein overexpressed in PDA tissue samples, can be used as a biomarker for PDA. Gal-1 levels were determined by ELISA in plasma from healthy controls and patients diagnosed with PDA, using three independent cohorts. Patients with chronic pancreatitis (CP) were also included in the study to analyze the potential of Gal-1 to discriminate between cancer and inflammatory process. Plasma Gal-1 levels were significantly increased in patients with PDA as compared to controls in all three cohorts. Gal-1 sensitivity and specificity values were similar to that of the CA19-9 biomarker (the only FDA-approved blood test biomarker for PDA), and the combination of Gal-1 and CA19-9 significantly improved their individual discriminatory powers. Moreover, high levels of Gal-1 were associated with lower survival in patients with non-resected tumors. Collectively, our data indicate a strong potential of using circulating Gal-1 levels as a biomarker for detection and prognostics of patients with PDA., This work was supported by grants from the Spanish Ministerio de Economía y Competitividad/ ISCIII-FEDER (PI14/00125; PI17/00199), the Carmen Delgado/ Miguel Pérez-Mateo AESPANC-ACANPAN 2016 grant and the “Generalitat de Catalunya” (2014/SGR/143 and 2017/ SGR/255) to P.N., ISCIII PI15/02101, EU TRANSCAN AC14/00033 and AECC grants to A.C., and grants from Instituto de Salud Carlos III/FEDER (PT17/0015/0011) and the “Xarxa de Bancs de tumors” sponsored by Pla Director d’Oncologia de Catalunya (XBTC). CAO received funding from the International PhD studies Fellowship “Créditos Beca Francisco José de Caldas” n° 529-2011 Colciencias, Colombia.

The authors wish to make the following corrections to this paper [...]

Trabajo presentado en la XVI Reunión de la Asociación Española de Pancreatología, celebrada en Bilbao (España), del 19 al 21 de septiembre de 2019, Background: Pancreatic ductal adenocarcinoma (PDA) is one of the most lethal tumor types, with extremely low survival rates due to late diagnosis and resistance to standard therapies. Identification of new molecular targets and biomarkers is therefore an urgent need to improve this pessimistic scenario. Here we analyze the potential use of galectin-1 (Gal1), a glycan-binding protein that is highly overexpressed in PDA stroma, as therapeutic target and biomarker for PDA. Methods: To study the potential use of Gal1 as therapeutic target, effects of Gal1 inhibition has been tested using genetically engineered mouse models (Gal1 knockout mice and a KRas-driven mouse model of PDA) and a human cell-based system. To analyze whether Gal1 can be used as PDA biomarker, its levels were determined by ELISA in plasma from healthy controls and patients diagnosed with PDA, using three independent cohorts Results: Both in mouse models and in a human setting, inhibition of Gal1 leads to impaired tumor formation and metastasis, through modulation of multiple events including proliferation, angiogenesis and immune response. In addition, we found that plasma Gal-1 levels were significantly increased in PDA patients as compared to controls and that high levels of Gal-1 were associated with lower survival in patients with non-resected tumors. Conclusions: Gal1 plays a key role in PDA development and progression, highlighting the high therapeutic value of Gal1 inhibition in PDA treatment. Moreover, detection of Gal1 circulating levels shows a strong potential to be used as a novel biomarker for detection and prognostics of PDA patients.

Background Widespread abdominal imaging has led to a substantial increase in the detection of incidentalomas. Currently, an increasing number of centers offer surveillance of the pancreas to individuals at high risk (IARs) of pancreatic ductal adenocarcinoma (PDAC). Objective The aims of this study were to evaluate the frequency and type of incidental findings in a magnetic resonance imaging (MRI)-based surveillance program for IARs for PDAC, and to discuss the benefit of detecting these lesions. Methods The outcome of MRI screening was reviewed in 568 individuals from three long-term pancreas surveillance programs conducted at three large European expert centers. All MRIs were studied in detail for the presence of incidental lesions. Results The most common lesions were liver cysts, renal cysts and liver hemangioma, which together comprised 75% of all lesions. Only five (0.9%) patients underwent surgery for a benign lesion. Cancer was detected in 11 patients (1.9%); early detection of tumors was beneficial in at least five cases. Conclusion The present study demonstrates that extrapancreatic incidentaloma is a common finding in IARs for PDAC, but rarely requires additional treatment. CDKN2A-p16-Leiden mutation carriers were the only patient group found to harbor a substantial number of cancers, and detection resulted in benefit in several cases.

Adenocarcinoma of the pancreas (PDAC) accounts for 2.4% of all cancers diagnosed and is the fourth leading cause of cancer death, with almost equal rates of incidence and mortality [1]. By 2030, pancreatic cancer is projected to be the second leading cause of cancer-related death [2], surpassing breast, prostate and colorectal cancer. The overall survival at 5 years of around 7.2% as the majority of patients present with advanced disease at diagnosis. Patients with localized disease are treated with surgery, with or without neoadjuvant chemotherapy/ radiotherapy, followed by adjuvant chemotherapy. The majority (around 80%) of patients are treated only with chemotherapy as they have an advanced disease. Patients are treated in the first line with gemcitabine-abraxane or Folfirinox and with Naliri plus 5FU in the second line. There have been few clinical advances in PDAC treatment over the last 20 years and chemotherapy is the only treatment option available for the majority of patients. These tumours are also resistant to many targeted therapies such as anti-EGFR therapy like cetuximab [3] due to the presence of a KRAS mutation in the majority of primary tumors. Personalized medicine strategies have not yet been established in pancreatic cancer as in other more common tumour types. Thus, novel anti-tumour strategies are an important clinical need in order to improve survival rates., AEI/FEDER, UE DPI 2017-90147-R., Intramural call for new research projects for clinical researchers and emerging research groups. IRYCIS. (2018/0240), Iberoamerican Network CYTED-DITECROD-218RT0545.

Mosaic loss of chromosome Y (mLOY) leading to gonosomal XY/XO commonly occurs during aging, particularly in smokers. We investigated whether mLOY was associated with non-hematological cancer in three prospective cohorts (8,679 cancer cases and 5,110 cancer-free controls) and genetic susceptibility to mLOY. Overall, mLOY was observed in 7% of men, and its prevalence increased with age (per-year odds ratio (OR) = 1.13, 95% confidence interval (CI) = 1.12-1.15; P2 × 10(-16)), reaching 18.7% among men over 80 years old. mLOY was associated with current smoking (OR = 2.35, 95% CI = 1.82-3.03; P = 5.55 × 10(-11)), but the association weakened with years after cessation. mLOY was not consistently associated with overall or specific cancer risk (for example, bladder, lung or prostate cancer) nor with cancer survival after diagnosis (multivariate-adjusted hazard ratio = 0.87, 95% CI = 0.73-1.04; P = 0.12). In a genome-wide association study, we observed the first example of a common susceptibility locus for genetic mosaicism, specifically mLOY, which maps to TCL1A at 14q32.13, marked by rs2887399 (OR = 1.55, 95% CI = 1.36-1.78; P = 1.37 × 10(-10)).

Objective Surveillance programmes are recommended for individuals at risk (IAR) of familial pancreatic cancer (FPC) to detect early pancreatic cancer (pancreatic ductal adenocarcinoma, PDAC). However, the age to begin screening and the optimal screening protocol remain to be determined. Methods IAR from non- CDKN2A FPC families underwent annual screening by MRI with endoscopic ultrasonography (EUS) in board-approved prospective screening programmes at three tertiary referral centres. The diagnostic yield according to age and different screening protocols was analysed. Results 253 IAR with a median age of 48 (25–81) years underwent screening with a median of 3 (1–11) screening visits during a median follow-up of 28 (1–152) months. 134 (53%) IAR revealed pancreatic lesions on imaging, mostly cystic (94%), on baseline or follow-up screening. Lesions were significantly more often identified in IAR above the age of 45 years (p

Exosome production from cancer-associated fibroblasts seems to be an important driver of tumor progression. We report the first in-depth biotype characterization of ncRNAs, analyzed by Next Generation Sequencing and Bioinformatics, expressed in established primary human normal and cancer-associated fibroblasts (CAFs) from cancer and normal mucosa tissues from 9 colorectal cancer patients, and/or packaged in their derived exosomes. Differential representation and enrichment analyses based on these ncRNAs revealed a significant number of differences between the ncRNA content of exosomes and the expression patterns of the normal and cancer-associated fibroblast cells. ncRNA regulatory elements are specifically packaged in CAF-derived exosomes, supporting a specific cross-talk between CAFs and colon cancer cells and/or other stromal cells, mediated by exosomes. These sncRNAs are potential biomarkers present in cancer-associated fibroblast-derived exosomes, which should thereby contribute to developing new non-invasive diagnostic, prognostic and predictive methods for clinical applications in management of cancer patients., This research is supported by PI12/02037, PI12/01635, PI15/02101, PI17/01847 and RD12/0036/0041 from the Instituto de Salud Carlos III (Plan Estatal de I + D + i 2013–2016) and cofinanced by the European Development Regional Fund “A way to achieve Europe” (ERDF); by “CIBER de Cáncer”, CB16/12/00273, CB16/12/00301, CB16/12/00446 and CB16/12/00291, from the Instituto de Salud Carlos III-FEDER; by the Fundación Científica AECC (a multifaceted approach to target pancreatic cancer); by SAF2010–20750, and SAF2016-76377-R from the Ministerio de Economía y Competitividad of SpainFEDER; by S2010/BMD-2344 from the Comunidad de Madrid; and by the Fundación Banco Santander. Cristina Peña is a recipient of a Miguel Servet Contract from the Instituto de Salud Carlos III.

The use of blood samples as liquid biopsy is a label-free method for cancer diagnosis that offers benefits over traditional invasive biopsy techniques. Cell sorting by acoustic waves offers a means to separate rare cells from blood samples based on their physical properties in a label-free, contactless and biocompatible manner. Herein, we describe a flow-through separation approach that provides an efficient separation of tumor cells (TCs) from white blood cells (WBCs) in a microfluidic device, "THINUS-Chip" (Thin-Ultrasonic-Separator-Chip), actuated by ultrasounds. We introduce for the first time the concept of plate acoustic waves (PAW) applied to acoustophoresis as a new strategy. It lies in the geometrical chip design: different to other microseparators based on either bulk acoustic waves (BAW) or surface waves (SAW, SSAW and tSAW), it allows the use of polymeric materials without restrictions in the frequency of work. We demonstrate its ability to perform high-throughput isolation of TCs from WBCs, allowing a recovery rate of 84%±8% of TCs with a purity higher than 80% and combined viability of 85% at a flow rate of 80 μL/min (4.8 mL/h). The THINUS-Chip performs cell fractionation with low-cost manufacturing processes, opening the door to possible easy printing fabrication., This work was partially supported by the Ministry of Economy and Competence of Spain (MINECO) under two projects: BIO2011-30535-C04-01: Development of a high-throughput ultrasonic technology for the isolation of circulating tumor cells from peripheral blood samples; and DPI2017-90147-R: Low intensity a Low Intensity ultrasounds for early diagnosis and modulation of tumor and stroma

Background The management of advanced neuroendocrine tumors (NETs) has recently changed. We assessed the activity of pazopanib after failure of other systemic treatments in advanced NETs. Methods This was a multicenter, open-label, phase II study evaluating pazopanib as a single agent in advanced NETs (PAZONET study). The clinical benefit rate (CBR) at 6 months was the primary end point. Translational correlation of radiological response and progression-free survival (PFS) with circulating and tissue biomarkers was also evaluated. Results A total of 44 patients were enrolled. Twenty-five patients (59.5%) were progression-free at 6 months (4 partial responses, 21 stable diseases) with a median PFS of 9.5 months [95% confidence interval (CI) 4.8–14.1]. The CBR varied according to prior therapy received, with 73%, 60% and 25% in patients treated with prior multitarget inhibitors, prior mTOR inhibitors and both agents, respectively. A nonsignificant increase in PFS was observed in patients presenting lower baseline circulating tumor cell (CTC) counts (9.1 versus 5.8 months; P = 0.22) and in those with decreased levels of soluble-vascular endothelial growth factor receptor-2 (sVEGFR-2) (12.6 versus 9.1 months; P = 0.067). A trend toward reduced survival was documented in patients with VEGFR3 rs307821 and rs307826 missense polymorphisms [hazard ratio (HR): 12.3; 95% CI 1.09–139.2; P = 0.042 and HR: 6.9; 95% CI 0.96–49.9; P = 0.055, respectively]. Conclusions Pazopanib showed clinical activity in patients with advanced NETs regardless of previous treatments. Additionally, CTCs, soluble-s VEFGR-2 and VEGFR3 gene polymorphisms constitute potential biomarkers for selecting patients for pazopanib (NCT01280201). Clinical trial number NCT01280201.

Pancreatic ductal adenocarcinoma is an aggressive malignancy with a 5-year survival rate of approximately 5%. The lack of established strategies for early detection contributes to this poor prognosis. Although several novel candidate biomarkers have been proposed for earlier diagnosis, none have been adopted into routine clinical use. In this review, the authors examine the challenges associated with finding new pancreatic cancer diagnostic biomarkers and explore why translation of biomarker research for patient benefit has thus far failed. The authors also review recent progress and highlight advances in the understanding of the biology of pancreatic cancer that may lead to improvements in biomarker detection and implementation.

e13517 Background: Glioblastoma (GB) is an aggressive tumor. Circulating Endothelial Cells (CECs) can be detected in peripheral blood and have been related to angiogenesis. CECs have an unclear prognostic value in patients with glioblastoma. The objective of this study was to quantify the presence and number of CECs in GB patients and determine its potential prognostic role. Methods: In this prospective, single center study, peripheral blood samples were obtained at the time of GB diagnosis. For the detection of CECs, 4 ml of blood was analyzed using the CellSearch system (Veridex). CEC blood samples were classified as CD146+, CD105+, CD45− and DAPI+. CEC detection was performed prior to surgery in a total of 26 patients and 22 patients with glioblastoma were included in the final analysis. We mesured progression free survival (PFS) and overall survival (OS). Results: Between 2014 and 2016, twenty two patients with histologically confirmed glioblastoma were studied. There were 14 males and 8 females with an average age of 63 years (range 42-81). A complete resection was achieved in 50% of cases, partial or subtotal resection in 31.8% and biopsy in 18%. The mean number of CECs was 59.3 cells/mL (range 0-954). Patients were classified into two groups depending on the number of CECs: group 1 had a CEC count below the mean and group 2 had a CEC count above the mean. A total of 17 patients (77.2%) were in group 1 and 5 patients (22.7%) were in group 2.The median OS was 14.4 months (range -0.36-33.36) for the entire study population. The median OS was 17.4 months (IC95% 11.47-23.37) for patients in group 1 and 8.12 months for patients in group 2 (IC 95% 4.38-11.85) p = 0.002. The median PFS was 8.94 m (IC95% 5.1-12.69) for group 1 and 3.95 for group 2 (IC 95% 2.91-4.98) p = 0.097. Conclusions: CECs can be detected in recently diagnosed GB patients. The present study demonstrates a possible prognostic value for CEC determination in GB cases at the time of diagnosis.