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Pancreatic Neoplasms: HELP
Articles by Christopher Heeschen
Based on 28 articles published since 2010
(Why 28 articles?)
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Between 2010 and 2020, C. Heeschen wrote the following 28 articles about Pancreatic Neoplasms.
 
+ Citations + Abstracts
Pages: 1 · 2
1 Review Concise Review: Stem Cells in Pancreatic Cancer: From Concept to Translation. 2015

Raj, Deepak / Aicher, Alexandra / Heeschen, Christopher. ·Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, United Kingdom. ·Stem Cells · Pubmed #26202953.

ABSTRACT: Pancreatic cancer stem cells (CSCs) have been first described in 2007 and since then have emerged as an intriguing entity of cancer cells with distinct functional features including self-renewal and exclusive in vivo tumorigenicity. The heterogeneous pancreatic CSC pool has been implicated in tumor propagation as well as metastatic spread. Clinically, the most important feature of CSCs is their strong resistance to standard chemotherapy, which results in fast disease relapse, even with today's more advanced chemotherapeutic regimens. Therefore, novel therapeutic strategies to most efficiently target pancreatic CSCs are being developed and their careful clinical translation should provide new avenues to eradicate this deadly disease.

2 Review Stem cells as the root of pancreatic ductal adenocarcinoma. 2012

Balic, Anamaria / Dorado, Jorge / Alonso-Gómez, Mercedes / Heeschen, Christopher. ·Clinical Research Programme, Stem Cells & Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. ·Exp Cell Res · Pubmed #22119145.

ABSTRACT: Emerging evidence suggests that stem cells play a crucial role not only in the generation and maintenance of different tissues, but also in the development and progression of malignancies. For the many solid cancers, it has now been shown that they harbor a distinct subpopulation of cancer cells that bear stem cell features and therefore, these cells are termed cancer stem cells (CSC) or tumor-propagating cells. CSC are exclusively tumorigenic and essential drivers for tumor progression and metastasis. Moreover, it has been shown that pancreatic ductal adenocarcinoma does not only contain one homogeneous population of CSC rather than diverse subpopulations that may have evolved during tumor progression. One of these populations is called migrating CSC and can be characterized by CXCR4 co-expression. Only these cells are capable of evading the primary tumor and traveling to distant sites such as the liver as the preferred site of metastatic spread. Clinically even more important, however, is the observation that CSC are highly resistant to chemo- and radiotherapy resulting in their relative enrichment during treatment and rapid relapse of disease. Many laboratories are now working on the further in-depth characterization of these cells, which may eventually allow for the identification of their Achilles heal and lead to novel treatment modalities for fighting this deadly disease.

3 Review Pancreatic cancer stem cells: new insights and perspectives. 2011

Dorado, Jorge / Lonardo, Enza / Miranda-Lorenzo, Irene / Heeschen, Christopher. ·Stem Cells and Cancer Group, Clinical Research Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. ·J Gastroenterol · Pubmed #21698355.

ABSTRACT: Since the identification of self-renewing cells in the hematopoietic system several decades ago, stem cells have changed the way we study biology and medicine. Solid tumors contain a distinct subpopulation of cells that have stem cell characteristics and are exclusively responsible for tumorigenicity. This discovery has led to the development of the stem cell concept of cancer, which proposes that a subpopulation of self-renewing tumor cells, also termed cancer stem cells, is responsible for tumorigenesis and metastasis. This contrasts with the stochastic model of tumor development, which holds that all tumor cells are capable of tumor initiation. Different subpopulations of cancer stem cells have been identified in pancreatic ductal adenocarcinoma, based on the use of combinations of surface markers that allow their isolation, propagation, and further characterization. Importantly, cancer stem cells are not only capable of self-renewal and differentiation, but may also confer virulence via immune system evasion and multidrug resistance, and potentially via vasculogenic mimicry and transition to migratory and metastasizing derivatives. Therapeutic targeting of this subset of cells and the pathways defining their virulence holds great promise for the development of more effective strategies for the amelioration and eradication of this most lethal form of cancer.

4 Review Pancreatic cancer stem cells - update and future perspectives. 2010

Lonardo, Enza / Hermann, Patrick C / Heeschen, Christopher. ·Clinical Research Programme, Spanish National Cancer Research Centre (CNIO), C/ Melchor Fernandez Almagro 3, Madrid, Spain. ·Mol Oncol · Pubmed #20580623.

ABSTRACT: Solid tumours are the most common cancers and represent a major therapeutic challenge. The cancer stem cell hypothesis is an attractive model to explain the functional heterogeneity commonly observed in solid tumours. It proposes a hierarchical organization of tumours, in which a subpopulation of stem cell-like cells sustains tumour growth, metastasis, and resistance to therapy. We will present the most recent advances in the cancer stem cell field, with particular emphasis on pancreatic cancer as one of the deadliest human tumours, and highlight open questions and caveats to be addressed in future studies. There is increasing evidence that solid tumours including pancreatic cancer are hierarchically organized and sustained by a distinct subpopulation of cancer stem cells. However, direct evidence for the validity of the cancer stem cell hypothesis in human pancreatic cancer remains controversial due to the limitations of xenograft models but supportive data are now emerging from mouse models using related or different sets of markers for the identification of murine cancer stem cells. Therefore, while the clinical relevance of cancer stem cells remains a fundamental issue for this rapidly emerging field, current findings clearly suggest that specific elimination of these cells is possible and therapeutically relevant. Targeting of signalling pathways that are of particular importance for the maintenance and the elimination of cancer stem cell as the proposed root of the tumour may lead to the development of novel treatment regimens for pancreatic cancer. Here we will review the current literature on pancreatic cancer stem cells and the future perspective of this rapidly emerging field.

5 Article Epigenetic reprogramming of primary pancreatic cancer cells counteracts their in vivo tumourigenicity. 2019

Khoshchehreh, Reyhaneh / Totonchi, Mehdi / Carlos Ramirez, Juan / Torres, Raul / Baharvand, Hossein / Aicher, Alexandra / Ebrahimi, Marzieh / Heeschen, Christopher. ·Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. · Department of Developmental Biology, University of Science and Culture, Tehran, Iran. · Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran. · VIVEBioTECH, San Sebastian, 20009, Spain. · Molecular Cytogenetics and Genome Editing Unit, Human Cancer Genetics Program, Centro Nacional de Investigaciones Oncológicas (CNIO), Madrid, 28029, Spain. · Josep Carreras Leukemia Research Institute and Department of Biomedicine, School of Medicine, University of Barcelona, Barcelona, 08036, Spain. · Gene and Stem Cell Therapy Program, Centenary Institute, the University of Sydney, Camperdown, 2050, NSW, Australia. aicher_a@yahoo.com. · Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, 28029, Madrid, Spain. aicher_a@yahoo.com. · Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Mebrahimi@royaninstitute.org. · Department of Developmental Biology, University of Science and Culture, Tehran, Iran. Mebrahimi@royaninstitute.org. · Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, 28029, Madrid, Spain. christopher.heeschen@icloud.com. ·Oncogene · Pubmed #31308488.

ABSTRACT: Pancreatic ductal adenocarcinoma (PDAC) arises through accumulation of multiple genetic alterations. However, cancer cells also acquire and depend on cancer-specific epigenetic changes. To conclusively demonstrate the crucial relevance of the epigenetic programme for the tumourigenicity of the cancer cells, we used cellular reprogramming technology to reverse these epigenetic changes. We reprogrammed human PDAC cultures using three different techniques - (1) lentivirally via induction of Yamanaka Factors (OSKM), (2) the pluripotency-associated gene OCT4 and the microRNA mir-302, or (3) using episomal vectors as a safer alternative without genomic integration. We found that induction with episomal vectors was the most efficient method to reprogram primary human PDAC cultures as well as primary human fibroblasts that served as positive controls. Successful reprogramming was evidenced by immunostaining, alkaline phosphatase staining, and real-time PCR. Intriguingly, reprogramming of primary human PDAC cultures drastically reduced their in vivo tumourigenicity, which appeared to be driven by the cells' enhanced differentiation and loss of stemness upon transplantation. Our study demonstrates that reprogrammed primary PDAC cultures are functionally distinct from parental PDAC cells resulting in drastically reduced tumourigenicity in vitro and in vivo. Thus, epigenetic alterations account at least in part for the tumourigenicity and aggressiveness of pancreatic cancer, supporting the notion that epigenetic modulators could be a suitable approach to improve the dismal outcome of patients with pancreatic cancer.

6 Article Pancreatic cancer stem cell proliferation is strongly inhibited by diethyldithiocarbamate-copper complex loaded into hyaluronic acid decorated liposomes. 2019

Marengo, Alessandro / Forciniti, Stefania / Dando, Ilaria / Dalla Pozza, Elisa / Stella, Barbara / Tsapis, Nicolas / Yagoubi, Najet / Fanelli, Giuseppina / Fattal, Elias / Heeschen, Christopher / Palmieri, Marta / Arpicco, Silvia. ·Department of Drug Science and Technology, University of Torino, Italy. · Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy. · Institut Galien Paris-Sud, CNRS, Université Paris-Sud, Université Paris-Saclay, 92296 Châtenay-Malabry, France. · EA 401, Matériaux et Santé, Université Paris-Sud, Université Paris-Saclay, 92296 Châtenay-Malabry, France. · Department of Ecological and Biological Sciences, University of Tuscia, Viterbo, Italy. · Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, UK. · Department of Neuroscience, Biomedicine and Movement, Biochemistry Section, University of Verona, Verona, Italy. Electronic address: marta.palmieri@univr.it. · Department of Drug Science and Technology, University of Torino, Italy. Electronic address: silvia.arpicco@unito.it. ·Biochim Biophys Acta Gen Subj · Pubmed #30267751.

ABSTRACT: BACKGROUND: Pancreatic cancer stem cells (CSCs) are responsible for resistance to standard therapy, metastatic potential, and disease relapse following treatments. The current therapy for pancreatic ductal adenocarcinoma (PDAC) preferentially targets the more differentiated cancer cell population, leaving CSCs as a cell source for tumor mass formation and recurrence. For this reason, there is an urgent need to improve current therapies and develop novel CSC-targeted therapeutic approaches. METHODS: Hyaluronic acid (HA) decorated liposomes, containing diethyldithiocarbamate‑copper (Cu(DDC) RESULTS: Liposomes showed high encapsulation efficiency and Cryo-TEM analysis revealed the presence of Cu(DDC) CONCLUSIONS: The obtained results show that the encapsulation of Cu(DDC) GENERAL SIGNIFICANCE: This paper describes for the first time the use of HA decorated liposomes containing Cu(DDC)

7 Article Switchable CAR-T cells mediate remission in metastatic pancreatic ductal adenocarcinoma. 2019

Raj, Deepak / Yang, Ming-Hsin / Rodgers, David / Hampton, Eric N / Begum, Julfa / Mustafa, Arif / Lorizio, Daniela / Garces, Irene / Propper, David / Kench, James G / Kocher, H M / Young, Travis S / Aicher, Alexandra / Heeschen, Christopher. ·Stem Cells in Cancer and Ageing, Barts Cancer Institute (BCI), Queen Mary University of London, London, UK. · Biologics, California Institute for Biomedical Research, La Jolla, California, USA. · Biological Service Unit, Barts Cancer Institute, London, UK. · Cancer and Inflammation, Barts Cancer Institute, London, UK. · Department of Tissue Pathology and Diagnostic Oncology, Royal Prince Alfred Hospital, Camperdown, New South Wales, Australia. · Director of the Barts Pancreatic Cancer Tissue Bank, Barts Cancer Institute (BCI), Queen Mary University of London, London, UK. · School of Medical Sciences, University of New South Wales, Sydney, New South Wales, Australia. ·Gut · Pubmed #30121627.

ABSTRACT: OBJECTIVE: Pancreatic ductal adenocarcinoma (PDAC) is a disease of unmet medical need. While immunotherapy with chimeric antigen receptor T (CAR-T) cells has shown much promise in haematological malignancies, their efficacy for solid tumours is challenged by the lack of tumour-specific antigens required to avoid on-target, off-tumour effects. Switchable CAR-T cells whereby activity of the CAR-T cell is controlled by dosage of a tumour antigen-specific recombinant Fab-based 'switch' to afford a fully tunable response may overcome this translational barrier. DESIGN: In this present study, we have used conventional and switchable CAR-T cells to target the antigen HER2, which is upregulated on tumour cells, but also present at low levels on normal human tissue. We used patient-derived xenograft models derived from patients with stage IV PDAC that mimic the most aggressive features of PDAC, including severe liver and lung metastases. RESULTS: Switchable CAR-T cells followed by administration of the switch directed against human epidermal growth factor receptor 2 (HER2)-induced complete remission in difficult-to-treat, patient-derived advanced pancreatic tumour models. Switchable HER2 CAR-T cells were as effective as conventional HER2 CAR-T cells in vivo testing a range of different CAR-T cell doses. CONCLUSION: These results suggest that a switchable CAR-T system is efficacious against aggressive and disseminated tumours derived from patients with advanced PDAC while affording the potential safety of a control switch.

8 Article The VAR2CSA malaria protein efficiently retrieves circulating tumor cells in an EpCAM-independent manner. 2018

Agerbæk, Mette Ø / Bang-Christensen, Sara R / Yang, Ming-Hsin / Clausen, Thomas M / Pereira, Marina A / Sharma, Shreya / Ditlev, Sisse B / Nielsen, Morten A / Choudhary, Swati / Gustavsson, Tobias / Sorensen, Poul H / Meyer, Tim / Propper, David / Shamash, Jonathan / Theander, Thor G / Aicher, Alexandra / Daugaard, Mads / Heeschen, Christopher / Salanti, Ali. ·Centre for Medical Parasitology at Department of Immunology and Microbiology, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, 2200, Copenhagen, Denmark. · Vancouver Prostate Centre, Vancouver, BC, V6H 3Z6, Canada. · Department of Urologic Sciences, University of British Columbia, Vancouver, BC, V5Z 1M9, Canada. · Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, EC1M 6BQ, United Kingdom. · Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, 11490, Taipei, Taiwan. · Department of Molecular Oncology, British Columbia Cancer Research Centre, Vancouver, BC, V5Z 1L3, Canada. · UCL Cancer Institute, University College London, London, WC1E 6BT, United Kingdom. · Department of Medical Oncology, Barts Health NHS, London, EC1A 7BE, United Kingdom. · Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, EC1M 6BQ, United Kingdom. c.heeschen@unsw.edu.au. · School of Medical Sciences, University of New South Wales, Sydney, NSW, 2052, Australia. c.heeschen@unsw.edu.au. · Centre for Medical Parasitology at Department of Immunology and Microbiology, University of Copenhagen and Department of Infectious Diseases, Copenhagen University Hospital, 2200, Copenhagen, Denmark. salanti@sund.ku.dk. ·Nat Commun · Pubmed #30115931.

ABSTRACT: Isolation of metastatic circulating tumor cells (CTCs) from cancer patients is of high value for disease monitoring and molecular characterization. Despite the development of many new CTC isolation platforms in the last decade, their isolation and detection has remained a challenge due to the lack of specific and sensitive markers. In this feasibility study, we present a method for CTC isolation based on the specific binding of the malaria rVAR2 protein to oncofetal chondroitin sulfate (ofCS). We show that rVAR2 efficiently captures CTCs from hepatic, lung, pancreatic, and prostate carcinoma patients with minimal contamination of peripheral blood mononuclear cells. Expression of ofCS is present on epithelial and mesenchymal cancer cells and is equally preserved during epithelial-mesenchymal transition of cancer cells. In 25 stage I-IV prostate cancer patient samples, CTC enumeration significantly correlates with disease stage. Lastly, rVAR2 targets a larger and more diverse population of CTCs compared to anti-EpCAM strategies.

9 Article TGF-β induces miR-100 and miR-125b but blocks let-7a through LIN28B controlling PDAC progression. 2018

Ottaviani, Silvia / Stebbing, Justin / Frampton, Adam E / Zagorac, Sladjana / Krell, Jonathan / de Giorgio, Alexander / Trabulo, Sara M / Nguyen, Van T M / Magnani, Luca / Feng, Hugang / Giovannetti, Elisa / Funel, Niccola / Gress, Thomas M / Jiao, Long R / Lombardo, Ylenia / Lemoine, Nicholas R / Heeschen, Christopher / Castellano, Leandro. ·Department of Surgery and Cancer, Division of Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine (ICTEM), London, W12 0NN, UK. · Department of Surgery and Cancer, HPB Surgical Unit, Imperial College, Hammersmith Hospital Campus, London, W12 0HS, UK. · Department of Surgery and Cancer, Division of Cancer, Imperial College London, Institute of Reproductive and Developmental Biology (IRDB), London, W12 0NN, UK. · Stem Cells & Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, 28028, Spain. · Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, EC1M 6BQ, UK. · Epigenetics and Genome Stability Team, The Institute of Cancer Research, 237 Fulham Road, London, SW3 6JB, UK. · Department of Medical Oncology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, 1081 HV, The Netherlands. · Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, Pisa, 56126, Italy. · Clinic for Gastroenterology, Endocrinology, Metabolism and Infectiology, Philipps-University Marburg, Marburg, 35037, Germany. · Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, London, EC1M 6BQ, UK. · Department of Surgery and Cancer, Division of Cancer, Imperial College London, Imperial Centre for Translational and Experimental Medicine (ICTEM), London, W12 0NN, UK. l.castellano@sussex.ac.uk. · University of Sussex, School of life Sciences, John Maynard Smith Building, Falmer, Brighton, BN1 9QG, UK. l.castellano@sussex.ac.uk. ·Nat Commun · Pubmed #29748571.

ABSTRACT: TGF-β/Activin induces epithelial-to-mesenchymal transition and stemness in pancreatic ductal adenocarcinoma (PDAC). However, the microRNAs (miRNAs) regulated during this response have remained yet undetermined. Here, we show that TGF-β transcriptionally induces MIR100HG lncRNA, containing miR-100, miR-125b and let-7a in its intron, via SMAD2/3. Interestingly, we find that although the pro-tumourigenic miR-100 and miR-125b accordingly increase, the amount of anti-tumourigenic let-7a is unchanged, as TGF-β also induces LIN28B inhibiting its maturation. Notably, we demonstrate that inactivation of miR-125b or miR-100 affects the TGF-β-mediated response indicating that these miRNAs are important TGF-β effectors. We integrate AGO2-RIP-seq with RNA-seq to identify the global regulation exerted by these miRNAs in PDAC cells. Transcripts targeted by miR-125b and miR-100 significantly overlap and mainly inhibit p53 and cell-cell junctions' pathways. Together, we uncover that TGF-β induces an lncRNA, whose encoded miRNAs, miR-100, let-7a and miR-125b play opposing roles in controlling PDAC tumourigenesis.

10 Article Multifunctionalized iron oxide nanoparticles for selective targeting of pancreatic cancer cells. 2017

Trabulo, Sara / Aires, Antonio / Aicher, Alexandra / Heeschen, Christopher / Cortajarena, Aitziber L. ·Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), 28029 Madrid, Spain; Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, EC1M 6BQ, UK. · CIC BiomaGUNE, Parque Tecnológico de San Sebastián, Paseo Miramón 182, Donostia-San Sebastián 20009, Spain; IMDEA Nanociencia and Nanobiotechnology Unit associated to Centro Nacional de Biotecnología (CNB-CSIC), Campus Universitario de Cantoblanco, Madrid 28049, Spain. · Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), 28029 Madrid, Spain. · Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), 28029 Madrid, Spain; Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, EC1M 6BQ, UK. Electronic address: c.heeschen@qmul.ac.uk. · CIC BiomaGUNE, Parque Tecnológico de San Sebastián, Paseo Miramón 182, Donostia-San Sebastián 20009, Spain; IMDEA Nanociencia and Nanobiotechnology Unit associated to Centro Nacional de Biotecnología (CNB-CSIC), Campus Universitario de Cantoblanco, Madrid 28049, Spain; Ikerbasque, Basque Foundation for Science, Mª Díaz de Haro 3, 48013 Bilbao, Spain. Electronic address: alcortajarena@cicbiomagune.es. ·Biochim Biophys Acta Gen Subj · Pubmed #28161480.

ABSTRACT: Nanomedicine nowadays offers novel solutions in cancer therapy by introducing multimodal treatments in one single formulation. In addition, nanoparticles act as nanocarriers changing the solubility, biodistribution and efficiency of the therapeutic molecules, thus generating more efficient treatments and reducing their side effects. To apply these novel therapeutic approaches, efforts are focused on the multi-functionalization of the nanoparticles and will open up new avenues to advanced combinational therapies. Pancreatic ductal adenocarcinoma (PDAC) is a cancer with unmet medical needs. Abundant expression of the anti-phagocytosis signal CD47 has also been observed on pancreatic cancer cells, in particular a subset of cancer stem cells (CSCs) responsible for resistance to standard therapy and metastatic potential. CD47 receptor is found on pancreatic cancer and highly expressed on CSCs, but not on normal pancreas. Inhibiting CD47 using monoclonal antibodies has been shown as an effective strategy to treat PDAC in vivo. However, CD47 inhibition effectively slowed tumor growth only in combination with Gemcitabine or Abraxane. In this work, we present the generation of multifunctionalized iron oxide magnetic nanoparticles (MNPs) that include the anti-CD47 antibody and the chemotherapeutic drug Gemcitabine in a single formulation. We demonstrate the in vitro efficacy of the formulation against CD47-positive pancreatic cancer cells. This article is part of a Special Issue entitled "Recent Advances in Bionanomaterials" Guest Editor: Dr. Marie-Louise Saboungi and Dr. Samuel D. Bader.

11 Article ER stress protein AGR2 precedes and is involved in the regulation of pancreatic cancer initiation. 2017

Dumartin, L / Alrawashdeh, W / Trabulo, S M / Radon, T P / Steiger, K / Feakins, R M / di Magliano, M P / Heeschen, C / Esposito, I / Lemoine, N R / Crnogorac-Jurcevic, T. ·Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, London, UK. · Centre for Stem Cells in Cancer &Ageing, Barts Cancer Institute, Queen Mary University of London, London, UK. · Institute of Pathology, Technische Universität München, Munich, Germany. · Department of Histopathology, Royal London Hospital, London, UK. · Department of Surgery, Cell and Developmental Biology, University of Michigan, Ann Arbor, USA. · Institute of Pathology, Heinrich-Heine-University of Düsseldorf, Germany. ·Oncogene · Pubmed #27941872.

ABSTRACT: The mechanisms of initiation of pancreatic ductal adenocarcinoma (PDAC) are still largely unknown. In the present study, we analysed the role of anterior gradient-2 (AGR2) in the earliest stages of pancreatic neoplasia. Immunohistochemical analysis of chronic pancreatitis (CP) and peritumoral areas in PDAC tissues showed that AGR2 was present in tubular complexes (TC) and early pancreatic intraepithelial neoplasia (PanINs). Moreover, AGR2 was also found in discrete subpopulations of non-transformed cells neighbouring these pre-neoplastic lesions. In primary cells derived from human patient-derived xenograft (PDX) model, flow-cytometry revealed that AGR2 was overexpressed in pancreatic cancer stem cells (CSC) compared with non-stem cancer cells. In LSL-Kras

12 Article DNMT1 Inhibition Reprograms Pancreatic Cancer Stem Cells via Upregulation of the miR-17-92 Cluster. 2016

Zagorac, Sladjana / Alcala, Sonia / Fernandez Bayon, Gustavo / Bou Kheir, Tony / Schoenhals, Matthieu / González-Neira, Anna / Fernandez Fraga, Mario / Aicher, Alexandra / Heeschen, Christopher / Sainz, Bruno. ·Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, United Kingdom. Stem Cells & Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Stem Cells & Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. Department of Biochemistry, Universidad Autónoma de Madrid, Instituto de Investigaciones Biomédicas "Alberto Sols" CSIC-UAM, Madrid, Spain. · Cancer Epigenetics Unit, Asturias Central University Hospital, Spanish Council for Scientific Research (CSIC), Oviedo, Spain. · Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, United Kingdom. · Human Genotyping-Cegen Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, United Kingdom. Stem Cells & Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. c.heeschen@qmul.ac.uk. ·Cancer Res · Pubmed #27261509.

ABSTRACT: Pancreatic ductal adenocarcinoma (PDAC) and other carcinomas are hierarchically organized, with cancer stem cells (CSC) residing at the top of the hierarchy, where they drive tumor progression, metastasis, and chemoresistance. As CSC and non-CSC share an identical genetic background, we hypothesize that differences in epigenetics account for the striking functional differences between these two cell populations. Epigenetic mechanisms, such as DNA methylation, play an important role in maintaining pluripotency and regulating the differentiation of stem cells, but the role of DNA methylation in pancreatic CSC is obscure. In this study, we investigated the genome-wide DNA methylation profile of PDAC CSC, and we determined the importance of DNA methyltransferases for CSC maintenance and tumorigenicity. Using high-throughput methylation analysis, we discovered that sorted CSCs have a higher level of DNA methylation, regardless of the heterogeneity or polyclonality of the CSC populations present in the tumors analyzed. Mechanistically, CSC expressed higher DNMT1 levels than non-CSC. Pharmacologic or genetic targeting of DNMT1 in CSCs reduced their self-renewal and in vivo tumorigenic potential, defining DNMT1 as a candidate CSC therapeutic target. The inhibitory effect we observed was mediated in part through epigenetic reactivation of previously silenced miRNAs, in particular the miR-17-92 cluster. Together, our findings indicate that DNA methylation plays an important role in CSC biology and also provide a rationale to develop epigenetic modulators to target CSC plasticity and improve the poor outcome of PDAC patients. Cancer Res; 76(15); 4546-58. ©2016 AACR.

13 Article MYC/PGC-1α Balance Determines the Metabolic Phenotype and Plasticity of Pancreatic Cancer Stem Cells. 2015

Sancho, Patricia / Burgos-Ramos, Emma / Tavera, Alejandra / Bou Kheir, Tony / Jagust, Petra / Schoenhals, Matthieu / Barneda, David / Sellers, Katherine / Campos-Olivas, Ramon / Graña, Osvaldo / Viera, Catarina R / Yuneva, Mariia / Sainz, Bruno / Heeschen, Christopher. ·Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London EC1M 6BQ, UK; Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain. Electronic address: p.sancho@qmul.ac.uk. · Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain. · Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London EC1M 6BQ, UK. · The Francis Crick Institute, Mill Hill Laboratories, The Ridgeway, London NW7 1AA, UK. · Spectroscopy and NMR Unit, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain. · Bioinformatics Unit and Structural Biology and Biocomputing Programme, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain. · Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London EC1M 6BQ, UK; Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain. Electronic address: c.heeschen@qmul.ac.uk. ·Cell Metab · Pubmed #26365176.

ABSTRACT: The anti-diabetic drug metformin targets pancreatic cancer stem cells (CSCs), but not their differentiated progenies (non-CSCs), which may be related to distinct metabolic phenotypes. Here we conclusively demonstrate that while non-CSCs were highly glycolytic, CSCs were dependent on oxidative metabolism (OXPHOS) with very limited metabolic plasticity. Thus, mitochondrial inhibition, e.g., by metformin, translated into energy crisis and apoptosis. However, resistant CSC clones eventually emerged during treatment with metformin due to their intermediate glycolytic/respiratory phenotype. Mechanistically, suppression of MYC and subsequent increase of PGC-1α were identified as key determinants for the OXPHOS dependency of CSCs, which was abolished in resistant CSC clones. Intriguingly, no resistance was observed for the mitochondrial ROS inducer menadione and resistance could also be prevented/reversed for metformin by genetic/pharmacological inhibition of MYC. Thus, the specific metabolic features of pancreatic CSCs are amendable to therapeutic intervention and could provide the basis for developing more effective therapies to combat this lethal cancer.

14 Article Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies. 2015

Lonardo, Enza / Cioffi, Michele / Sancho, Patricia / Crusz, Shanthini / Heeschen, Christopher. ·Stem Cells & Cancer Group, Molecular Pathology Program, Spanish National Cancer Research Center; Institute for Research in Biomedicine (IRB Barcelona); enza.lonardo@irbbarcelona.org. · Stem Cells & Cancer Group, Molecular Pathology Program, Spanish National Cancer Research Center. · Stem Cells & Cancer Group, Molecular Pathology Program, Spanish National Cancer Research Center; Center for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London. · Center for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London. · Stem Cells & Cancer Group, Molecular Pathology Program, Spanish National Cancer Research Center; Center for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London; cheeschen@cnio.es. ·J Vis Exp · Pubmed #26132091.

ABSTRACT: Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

15 Article The miR-17-92 cluster counteracts quiescence and chemoresistance in a distinct subpopulation of pancreatic cancer stem cells. 2015

Cioffi, Michele / Trabulo, Sara M / Sanchez-Ripoll, Yolanda / Miranda-Lorenzo, Irene / Lonardo, Enza / Dorado, Jorge / Reis Vieira, Catarina / Ramirez, Juan Carlos / Hidalgo, Manuel / Aicher, Alexandra / Hahn, Stephan / Sainz, Bruno / Heeschen, Christopher. ·Stem Cells & Cancer Group, CNIO, Madrid, Spain. · Stem Cells & Cancer Group, CNIO, Madrid, Spain Viral Vector Unit, Spanish National Cardiovascular Research Centre (CNIC), Madrid, Spain. · Viral Vector Unit, Spanish National Cardiovascular Research Centre (CNIC), Madrid, Spain. · Gastrointestinal Cancer Clinical Research Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Department of Molecular Gastrointestinal Oncology, Ruhr-University Bochum, D-44801 Bochum, Germany. · Stem Cells & Cancer Group, CNIO, Madrid, Spain Barts Cancer Institute, Centre for Stem Cells in Cancer & Ageing, Queen Mary University of London, London, UK. ·Gut · Pubmed #25887381.

ABSTRACT: OBJECTIVE: Cancer stem cells (CSCs) represent the root of many solid cancers including pancreatic ductal adenocarcinoma, are highly chemoresistant and represent the cellular source for disease relapse. However the mechanisms involved in these processes still need to be fully elucidated. Understanding the mechanisms implicated in chemoresistance and metastasis of pancreatic cancer is critical to improving patient outcomes. DESIGN: Micro-RNA (miRNA) expression analyses were performed to identify functionally defining epigenetic signatures in pancreatic CSC-enriched sphere-derived cells and gemcitabine-resistant pancreatic CSCs. RESULTS: We found the miR-17-92 cluster to be downregulated in chemoresistant CSCs versus non-CSCs and demonstrate its crucial relevance for CSC biology. In particular, overexpression of miR-17-92 reduced CSC self-renewal capacity, in vivo tumourigenicity and chemoresistance by targeting multiple NODAL/ACTIVIN/TGF-β1 signalling cascade members as well as directly inhibiting the downstream targets p21, p57 and TBX3. Overexpression of miR-17-92 translated into increased CSC proliferation and their eventual exhaustion via downregulation of p21 and p57. Finally, the translational impact of our findings could be confirmed in preclinical models for pancreatic cancer. CONCLUSIONS: Our findings therefore identify the miR-17-92 cluster as a functionally determining family of miRNAs in CSCs, and highlight the putative potential of developing modulators of this cluster to overcome drug resistance in pancreatic CSCs.

16 Article Microenvironmental hCAP-18/LL-37 promotes pancreatic ductal adenocarcinoma by activating its cancer stem cell compartment. 2015

Sainz, Bruno / Alcala, Sonia / Garcia, Elena / Sanchez-Ripoll, Yolanda / Azevedo, Maria M / Cioffi, Michele / Tatari, Marianthi / Miranda-Lorenzo, Irene / Hidalgo, Manuel / Gomez-Lopez, Gonzalo / Cañamero, Marta / Erkan, Mert / Kleeff, Jörg / García-Silva, Susana / Sancho, Patricia / Hermann, Patrick C / Heeschen, Christopher. ·Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain Department of Preventive Medicine, Public Health and Microbiology, Universidad Autónoma de Madrid, Madrid, Spain. · Molecular Diagnostics Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain Pathology Department, Hospital Universitario Fundacion Alcorcon, Madrid, Spain. · Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Gastrointestinal Cancer Clinical Research Unit, Clinical Research Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Bioinformatics Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Histopathology Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Department of Surgery, Klinikum rechts der Isar, Technical University, Munich, Germany Koc University School of Medicine, Istanbul, Turkey. · Department of Surgery, Klinikum rechts der Isar, Technical University, Munich, Germany. · Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain Deptartment of Internal Medicine I, Ulm University, Ulm, Germany. · Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, London, UK. ·Gut · Pubmed #25841238.

ABSTRACT: OBJECTIVES: The tumour stroma/microenvironment not only provides structural support for tumour development, but more importantly it provides cues to cancer stem cells (CSCs) that regulate their self-renewal and metastatic potential. This is certainly true for pancreatic ductal adenocarcinomas (PDAC), where tumour-associated fibroblasts, pancreatic stellate cells and immune cells create an abundant paracrine niche for CSCs via microenvironment-secreted factors. Thus understanding the role that tumour stroma cells play in PDAC development and CSC biology is of utmost importance. DESIGN: Microarray analyses, tumour microarray immunohistochemical assays, in vitro co-culture experiments, recombinant protein treatment approaches and in vivo intervention studies were performed to understand the role that the immunomodulatory cationic antimicrobial peptide 18/LL-37 (hCAP-18/LL-37) plays in PDAC biology. RESULTS: We found that hCAP-18/LL-37 was strongly expressed in the stroma of advanced primary and secondary PDAC tumours and is secreted by immune cells of the stroma (eg, tumour-associated macrophages) in response to tumour growth factor-β1 and particularly CSC-secreted Nodal/ActivinA. Treatment of pancreatic CSCs with recombinant LL-37 increased pluripotency-associated gene expression, self-renewal, invasion and tumourigenicity via formyl peptide receptor 2 (FPR2)- and P2X purinoceptor 7 receptor (P2X7R)-dependent mechanisms, which could be reversed by inhibiting these receptors. Importantly, in a genetically engineered mouse model of K-Ras-driven pancreatic tumourigenesis, we also showed that tumour formation was inhibited by either reconstituting these mice with bone marrow from cathelicidin-related antimicrobial peptide (ie, murine homologue of hCAP-18/LL-37) knockout mice or by pharmacologically inhibiting FPR2 and P2X7R. CONCLUSIONS: Thus, hCAP-18/LL-37 represents a previously unrecognised PDAC microenvironment factor that plays a critical role in pancreatic CSC-mediated tumourigenesis.

17 Article Inhibition of CD47 Effectively Targets Pancreatic Cancer Stem Cells via Dual Mechanisms. 2015

Cioffi, Michele / Trabulo, Sara / Hidalgo, Manuel / Costello, Eithne / Greenhalf, William / Erkan, Mert / Kleeff, Joerg / Sainz, Bruno / Heeschen, Christopher. ·Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. Centre for Stem Cells in Cancer and Ageing, Barts Cancer Institute, A CR-UK Centre of Excellence, Queen Mary University of London, United Kingdom. · Gastrointestinal Cancer Clinical Research Unit, Clinical Research Programme, CNIO, Madrid, Spain. · Liverpool Cancer Research UK Centre, University of Liverpool, Liverpool, United Kingdom. · Department of Surgery, Technical University Munich, Munich, Germany. Koc University School of Medicine, Instanbul, Turkey. · Department of Surgery, Technical University Munich, Munich, Germany. · Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. Centre for Stem Cells in Cancer and Ageing, Barts Cancer Institute, A CR-UK Centre of Excellence, Queen Mary University of London, United Kingdom. c.heeschen@qmul.ac.uk. ·Clin Cancer Res · Pubmed #25717063.

ABSTRACT: PURPOSE: Pancreatic ductal adenocarcinoma (PDAC) is a cancer of the exocrine pancreas with unmet medical need and is strongly promoted by tumor-associated macrophages (TAM). The presence of TAMs is associated with poor clinical outcome, and their overall role, therefore, appears to be protumorigenic. The "don't eat me" signal CD47 on cancer cells communicates to the signal regulatory protein-α on macrophages and prevents their phagocytosis. Thus, inhibition of CD47 may offer a new opportunity to turn TAMs against PDAC cells, including cancer stem cells (CSC), as the exclusively tumorigenic population. EXPERIMENTAL DESIGN: We studied in vitro and in vivo the effects of CD47 inhibition on CSCs using a large set of primary pancreatic cancer (stem) cells as well as xenografts of primary human PDAC tissue. RESULTS: CD47 was highly expressed on CSCs, but not on other nonmalignant cells in the pancreas. Targeting CD47 efficiently enhanced phagocytosis of a representative set of primary human pancreatic cancer (stem) cells and, even more intriguingly, also directly induced their apoptosis in the absence of macrophages during long-term inhibition of CD47. In patient-derived xenograft models, CD47 targeting alone did not result in relevant slowing of tumor growth, but the addition of gemcitabine or Abraxane resulted in sustained tumor regression and prevention of disease relapse long after discontinuation of treatment. CONCLUSIONS: These data are consistent with efficient in vivo targeting of CSCs, and strongly suggest that CD47 inhibition could be a novel adjuvant treatment strategy for PDAC independent of underlying and highly variable driver mutations.

18 Article ISG15 is a critical microenvironmental factor for pancreatic cancer stem cells. 2014

Sainz, Bruno / Martín, Beatriz / Tatari, Marianthi / Heeschen, Christopher / Guerra, Susana. ·Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. Department of Preventive Medicine, Public Health and Microbiology, Universidad Autónoma, Madrid, Spain. bruno.sainz@uam.es susana.guerra@uam.es. · Department of Preventive Medicine, Public Health and Microbiology, Universidad Autónoma, Madrid, Spain. · Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. Barts Cancer Institute, Queen Mary University of London, London, United Kingdom. · Department of Preventive Medicine, Public Health and Microbiology, Universidad Autónoma, Madrid, Spain. bruno.sainz@uam.es susana.guerra@uam.es. ·Cancer Res · Pubmed #25368022.

ABSTRACT: Cancer stem cells (CSC) are thought to play a major role in the development and metastatic progression of pancreatic ductal adenocarcinoma (PDAC), one of the deadliest solid tumors. Likewise, the tumor microenvironment contributes critical support in this setting, including from tumor stromal cells and tumor-associated macrophages (TAM) that contribute structural and paracrine-mediated supports, respectively. Here, we show that TAMs secrete the IFN-stimulated factor ISG15, which enhances CSC phenotypes in PDAC in vitro and in vivo. ISG15 was preferentially and highly expressed by TAM present in primary PDAC tumors resected from patients. ISG15 was secreted by macrophages in response to secretion of IFNβ by CSC, thereby reinforcing CSC self-renewal, invasive capacity, and tumorigenic potential. Overall, our work demonstrates that ISG15 is a previously unrecognized support factor for CSC in the PDAC microenvironment with a key role in pathogenesis and progression.

19 Article Intracellular autofluorescence: a biomarker for epithelial cancer stem cells. 2014

Miranda-Lorenzo, Irene / Dorado, Jorge / Lonardo, Enza / Alcala, Sonia / Serrano, Alicia G / Clausell-Tormos, Jenifer / Cioffi, Michele / Megias, Diego / Zagorac, Sladjana / Balic, Anamaria / Hidalgo, Manuel / Erkan, Mert / Kleeff, Joerg / Scarpa, Aldo / Sainz, Bruno / Heeschen, Christopher. ·Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Melanoma Group, Molecular Pathology Programme, CNIO, Madrid, Spain. · Confocal Microscopy Unit, Biotechnology Programme, CNIO, Madrid, Spain. · Gastrointestinal Cancer Clinical Research Unit, Clinical Research Programme, CNIO, Madrid, Spain. · Chirurgische Klinik, Technische Universität München, Munich, Germany. · Università degli Studi di Verona, Centro Ricerca Applicata ARC-NET, Verona, Italy. · 1] Stem Cells and Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. [2] Centre for Stem Cells in Cancer &Ageing, Barts Cancer Institute, Queen Mary University of London, London, UK. ·Nat Methods · Pubmed #25262208.

ABSTRACT: Cancer stem cells (CSCs) are thought to drive tumor growth, metastasis and chemoresistance. Although surface markers such as CD133 and CD44 have been successfully used to isolate CSCs, their expression is not exclusively linked to the CSC phenotype and is prone to environmental alteration. We identified cells with an autofluorescent subcellular compartment that exclusively showed CSC features across different human tumor types. Primary tumor-derived autofluorescent cells did not overlap with side-population (SP) cells, were enriched in sphere culture and during chemotherapy, strongly expressed pluripotency-associated genes, were highly metastatic and showed long-term in vivo tumorigenicity, even at the single-cell level. Autofluorescence was due to riboflavin accumulation in membrane-bounded cytoplasmic structures bearing ATP-dependent ABCG2 transporters. In summary, we identified and characterized an intrinsic autofluorescent phenotype in CSCs of diverse epithelial cancers and used this marker to isolate and characterize these cells.

20 Article Nicotine promotes initiation and progression of KRAS-induced pancreatic cancer via Gata6-dependent dedifferentiation of acinar cells in mice. 2014

Hermann, Patrick C / Sancho, Patricia / Cañamero, Marta / Martinelli, Paola / Madriles, Francesc / Michl, Patrick / Gress, Thomas / de Pascual, Ricardo / Gandia, Luis / Guerra, Carmen / Barbacid, Mariano / Wagner, Martin / Vieira, Catarina R / Aicher, Alexandra / Real, Francisco X / Sainz, Bruno / Heeschen, Christopher. ·Stem Cells and Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Comparative Pathology Core Unit, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Epithelial Carcinogenesis Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Department of Gastroenterology, Endocrinology, Metabolism and Infectiology, University of Marburg, Marburg, Germany. · Instituto Teófilo Hernando, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain. · Experimental Oncology Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. · Department of Internal Medicine I, Ulm University, Ulm, Germany. · Stem Cells and Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. Electronic address: bruno.sainz@uam.es. · Stem Cells and Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain; Centre for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London, UK. Electronic address: c.heeschen@qmul.ac.uk. ·Gastroenterology · Pubmed #25127677.

ABSTRACT: BACKGROUND & AIMS: Although smoking is a leading risk factor for pancreatic ductal adenocarcinoma (PDAC), little is known about the mechanisms by which smoking promotes initiation or progression of PDAC. METHODS: We studied the effects of nicotine administration on pancreatic cancer development in Kras(+/LSLG12Vgeo);Elas-tTA/tetO-Cre (Ela-KRAS) mice, Kras(+/LSLG12D);Trp53+/LSLR172H;Pdx-1-Cre (KPC) mice (which express constitutively active forms of KRAS), and C57/B6 mice. Mice were given nicotine for up to 86 weeks to produce blood levels comparable with those of intermediate smokers. Pancreatic tissues were collected and analyzed by immunohistochemistry and reverse transcriptase polymerase chain reaction; cells were isolated and assayed for colony and sphere formation and gene expression. The effects of nicotine were also evaluated in primary pancreatic acinar cells isolated from wild-type, nAChR7a(-/-), Trp53(-/-), and Gata6(-/-);Trp53(-/-) mice. We also analyzed primary PDAC cells that overexpressed GATA6 from lentiviral expression vectors. RESULTS: Administration of nicotine accelerated transformation of pancreatic cells and tumor formation in Ela-KRAS and KPC mice. Nicotine induced dedifferentiation of acinar cells by activating AKT-ERK-MYC signaling; this led to inhibition of Gata6 promoter activity, loss of GATA6 protein, and subsequent loss of acinar differentiation and hyperactivation of oncogenic KRAS. Nicotine also promoted aggressiveness of established tumors as well as the epithelial-mesenchymal transition, increasing numbers of circulating cancer cells and their dissemination to the liver, compared with mice not exposed to nicotine. Nicotine induced pancreatic cells to acquire gene expression patterns and functional characteristics of cancer stem cells. These effects were markedly attenuated in K-Ras(+/LSL-G12D);Trp53(+/LSLR172H);Pdx-1-Cre mice given metformin. Metformin prevented nicotine-induced pancreatic carcinogenesis and tumor growth by up-regulating GATA6 and promoting differentiation toward an acinar cell program. CONCLUSIONS: In mice, nicotine promotes pancreatic carcinogenesis and tumor development via down-regulation of Gata6 to induce acinar cell dedifferentiation.

21 Article Chloroquine targets pancreatic cancer stem cells via inhibition of CXCR4 and hedgehog signaling. 2014

Balic, Anamaria / Sørensen, Morten Dræby / Trabulo, Sara Maria / Sainz, Bruno / Cioffi, Michele / Vieira, Catarina R / Miranda-Lorenzo, Irene / Hidalgo, Manuel / Kleeff, Joerg / Erkan, Mert / Heeschen, Christopher. ·Authors' Affiliations: Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO); · Authors' Affiliations: Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO); Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark; · Authors' Affiliations: Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO); Barts Cancer Institute, Queen Mary University of London, London, United Kingdom. · Gastrointestinal Cancer Clinical Research Unit, Clinical Research Programme, CNIO, Madrid, Spain; · Department of Surgery, Klinikum Rechts der Isar, Technical University, Munich, Germany; and. · Authors' Affiliations: Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO); Barts Cancer Institute, Queen Mary University of London, London, United Kingdom c.heeschen@qmul.ac.uk. ·Mol Cancer Ther · Pubmed #24785258.

ABSTRACT: Pancreatic ductal adenocarcinoma is one of the deadliest carcinomas and is characterized by highly tumorigenic and metastatic cancer stem cells (CSC). CSCs evade available therapies, which preferentially target highly proliferative and more differentiated progenies, leaving behind CSCs as a putative source for disease relapse. Thus, to identify potentially more effective treatment regimens, we screened established and new compounds for their ability to eliminate CSCs in primary pancreatic cancer (stem) cells in vitro and corresponding patient-derived pancreatic cancer tissue xenografts in vivo. Intriguingly, we found that in vitro treatment with the antimalarial agent chloroquine significantly decreased CSCs, translating into diminished in vivo tumorigenicity and invasiveness in a large panel of pancreatic cancers. In vivo treatment in combination with gemcitabine was capable of more effectively eliminating established tumors and improved overall survival. The inhibitory effect of chloroquine was not related to inhibition of autophagy, but was due to inhibition of CXCL12/CXCR4 signaling, resulting in reduced phosphorylation of ERK and STAT3. Furthermore, chloroquine showed potent inhibition of hedgehog signaling by decreasing the production of Smoothened, translating into a significant reduction in sonic hedgehog-induced chemotaxis and downregulation of downstream targets in CSCs and the surrounding stroma. Our study demonstrates that via to date unreported effects, chloroquine is an effective adjuvant therapy to chemotherapy, offering more efficient tumor elimination and improved cure rates. Chloroquine should be further explored in the clinical setting as its success may help to more rapidly improve the poor prognosis of patients with pancreatic cancer.

22 Article Metformin targets the metabolic achilles heel of human pancreatic cancer stem cells. 2013

Lonardo, Enza / Cioffi, Michele / Sancho, Patricia / Sanchez-Ripoll, Yolanda / Trabulo, Sara Maria / Dorado, Jorge / Balic, Anamaria / Hidalgo, Manuel / Heeschen, Christopher. ·Stem Cells & Cancer Group, Molecular Pathology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. ·PLoS One · Pubmed #24204632.

ABSTRACT: Pancreatic ductal adenocarcinomas contain a subset of exclusively tumorigenic cancer stem cells (CSCs), which are capable of repopulating the entire heterogeneous cancer cell populations and are highly resistant to standard chemotherapy. Here we demonstrate that metformin selectively ablated pancreatic CSCs as evidenced by diminished expression of pluripotency-associated genes and CSC-associated surface markers. Subsequently, the ability of metformin-treated CSCs to clonally expand in vitro was irreversibly abrogated by inducing apoptosis. In contrast, non-CSCs preferentially responded by cell cycle arrest, but were not eliminated by metformin treatment. Mechanistically, metformin increased reactive oxygen species production in CSC and reduced their mitochondrial transmembrane potential. The subsequent induction of lethal energy crisis in CSCs was independent of AMPK/mTOR. Finally, in primary cancer tissue xenograft models metformin effectively reduced tumor burden and prevented disease progression; if combined with a stroma-targeting smoothened inhibitor for enhanced tissue penetration, while gemcitabine actually appeared dispensable.

23 Article Multimodal Treatment Eliminates Cancer Stem Cells and Leads to Long-Term Survival in Primary Human Pancreatic Cancer Tissue Xenografts. 2013

Hermann, Patrick C / Trabulo, Sara M / Sainz, Bruno / Balic, Anamaria / Garcia, Elena / Hahn, Stephan A / Vandana, Mallaredy / Sahoo, Sanjeeb K / Tunici, Patrizia / Bakker, Annette / Hidalgo, Manuel / Heeschen, Christopher. ·Stem Cells and Cancer Group, Clinical Research Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. ·PLoS One · Pubmed #23825539.

ABSTRACT: PURPOSE: In spite of intense research efforts, pancreatic ductal adenocarcinoma remains one of the most deadly malignancies in the world. We and others have previously identified a subpopulation of pancreatic cancer stem cells within the tumor as a critical therapeutic target and additionally shown that the tumor stroma represents not only a restrictive barrier for successful drug delivery, but also serves as a paracrine niche for cancer stem cells. Therefore, we embarked on a large-scale investigation on the effects of combining chemotherapy, hedgehog pathway inhibition, and mTOR inhibition in a preclinical mouse model of pancreatic cancer. EXPERIMENTAL DESIGN: Prospective and randomized testing in a set of almost 200 subcutaneous and orthotopic implanted whole-tissue primary human tumor xenografts. RESULTS: The combined targeting of highly chemoresistant cancer stem cells as well as their more differentiated progenies, together with abrogation of the tumor microenvironment by targeting the stroma and enhancing tissue penetration of the chemotherapeutic agent translated into significantly prolonged survival in preclinical models of human pancreatic cancer. Most pronounced therapeutic effects were observed in gemcitabine-resistant patient-derived tumors. Intriguingly, the proposed triple therapy approach could be further enhanced by using a PEGylated formulation of gemcitabine, which significantly increased its bioavailability and tissue penetration, resulting in a further improved overall outcome. CONCLUSIONS: This multimodal therapeutic strategy should be further explored in the clinical setting as its success may eventually improve the poor prognosis of patients with pancreatic ductal adenocarcinoma.

24 Article Stem cells & pancreatic cancer. 2013

García-Silva, Susana / Frias-Aldeguer, Javier / Heeschen, Christopher. ·Stem Cells & Cancer Group, Clinical Research Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain. ·Pancreatology · Pubmed #23561967.

ABSTRACT: It is now well established that human pancreatic ductal adenocarcinoma (PDAC) contains a subset of cells with self-renewal capabilities and subsequent exclusive in vivo tumorigenic capacity as assessed by limiting dilution tumorigenic transplantation assays into immunodeficient mice. These cells are considered pancreatic cancer stem cells (CSCs) and are able to form tumors indistinguishable from parental ones. Furthermore they display strong chemotherapy resistance and are implicated in tumor relapses and metastatic spread. Important next steps for advancing the field of pancreatic CSC research include the identification and characterization of CSCs in the unperturbed in vivo setting. This has been achieved just recently for other solid tumors such as glioblastoma using clonal analysis after lineage tracing in mice [1]. In vivo imaging of CSCs during tumor development should not only provide new insights into the in vivo features of CSCs, but also help to further unravel the influence of the stroma on CSC biology. Comprehensive studies of the tumor heterogeneity with respect to the coexistence of different clones potentially generated by distinct population of CSCs that are evolving by stochastic cell fate decisions may actually unite the CSC concept and the model of clonal evolution for pancreatic cancer. Eventually, the design of specific therapies against CSCs should open new alleys to improve survival of patients with PDAC. Combined therapies targeting CSCs and their progenies as well as the supportive stroma may represent the most promising approach for the future treatment of patients with PDAC.

25 Article Metastatic cancer stem cells--quo vadis? 2013

Hermann, Patrick C / Heeschen, Christopher. ·Spanish National Cancer Research Centre (CNIO), Madrid, Spain. ·Clin Chem · Pubmed #23288488.

ABSTRACT: -- No abstract --

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