Pancreatic Cancer-Induced Neutrophil Extracellular Traps: A Potential Contributor to Cancer-Associated Thrombosis

Pancreatic cancer (PaCa) is a highly metastatic cancer, and patients are at high risk of developing venous thromboembolism (VTE). Neutrophil extracellular traps (NETs) have been associated with cancer metastasis and cancer-associated thrombosis, but the ability of cancer to stimulate NET release is not known. The release of NETs has been shown to be a slow process and requires reactive oxygen species (ROS) production. Studies suggest that activated platelets are important mediators in the release. Here, we show that PaCa cells can stimulate the rapid release of NETs, independently of ROS production. We further assessed the role of platelets in PaCa-induced NETs and observed a trend of increased the NET release by PaCa-primed platelets. Additionally, NETs promoted thrombus formation under venous shear stress ex vivo. Taken together, our results suggest that PaCa-induced NETs can contribute to the high risk of venous thromboembolism development in PaCa patients, and reveal NETs as a potential therapeutic target.

[1]  C. van Kooten,et al.  A novel method for high-throughput detection and quantification of neutrophil extracellular traps reveals ROS-independent NET release with immune complexes. , 2016, Autoimmunity reviews.

[2]  A. Giatromanolaki,et al.  Gradient Infiltration of Neutrophil Extracellular Traps in Colon Cancer and Evidence for Their Involvement in Tumour Growth , 2016, PloS one.

[3]  J. Bae,et al.  Tumor-Associated Macrophages and Neutrophils in Tumor Microenvironment , 2016, Mediators of inflammation.

[4]  R. Gómez,et al.  Mediators and molecular pathways involved in the regulation of neutrophil extracellular trap formation mediated by activated platelets , 2016, Journal of leukocyte biology.

[5]  N. C. Rochael,et al.  Classical ROS-dependent and early/rapid ROS-independent release of Neutrophil Extracellular Traps triggered by Leishmania parasites , 2015, Scientific Reports.

[6]  A. Andrén-sandberg,et al.  Pancreatic cancer and thromboembolic disease, 150 years after Trousseau. , 2015, Hepatobiliary surgery and nutrition.

[7]  Pieter Wesseling,et al.  RNA-Seq of Tumor-Educated Platelets Enables Blood-Based Pan-Cancer, Multiclass, and Molecular Pathway Cancer Diagnostics , 2015, Cancer cell.

[8]  P. Metharom,et al.  Thrombin-induced reactive oxygen species generation in platelets: A novel role for protease-activated receptor 4 and GPIbα , 2015, Redox biology.

[9]  M. Czech,et al.  NETs and traps delay wound healing in diabetes , 2015, Trends in Endocrinology & Metabolism.

[10]  Julia Femel,et al.  Neutrophil Extracellular Traps Accumulate in Peripheral Blood Vessels and Compromise Organ Function in Tumor-Bearing Animals. , 2015, Cancer research.

[11]  J. Spicer,et al.  Simplified Human Neutrophil Extracellular Traps (NETs) Isolation and Handling. , 2015, Journal of visualized experiments : JoVE.

[12]  Ronit Vogt Sionov,et al.  Phenotypic diversity and plasticity in circulating neutrophil subpopulations in cancer. , 2015, Cell reports.

[13]  M. Lotze,et al.  The Receptor for Advanced Glycation End Products (RAGE) Enhances Autophagy and Neutrophil Extracellular Traps in Pancreatic Cancer , 2015, Cancer Gene Therapy.

[14]  I. Lizasoain,et al.  Neutrophils scan for activated platelets to initiate inflammation , 2014, Science.

[15]  A. Maseri,et al.  Activated platelets present high mobility group box 1 to neutrophils, inducing autophagy and promoting the extrusion of neutrophil extracellular traps , 2014, Journal of thrombosis and haemostasis : JTH.

[16]  R. Hynes,et al.  Platelets guide the formation of early metastatic niches , 2014, Proceedings of the National Academy of Sciences.

[17]  A. Buser,et al.  Enhanced neutrophil extracellular trap generation in rheumatoid arthritis: analysis of underlying signal transduction pathways and potential diagnostic utility , 2014, Arthritis Research & Therapy.

[18]  Hyeyoung Kim,et al.  Oxidative Stress and Cytokines in the Pathogenesis of Pancreatic Cancer , 2014, Journal of cancer prevention.

[19]  H. Malech,et al.  Release of Cystic Fibrosis Airway Inflammatory Markers from Pseudomonas aeruginosa–Stimulated Human Neutrophils Involves NADPH Oxidase-Dependent Extracellular DNA Trap Formation , 2014, The Journal of Immunology.

[20]  Paul Kubes,et al.  Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis. , 2013, The Journal of clinical investigation.

[21]  A. Walker,et al.  Incidence of venous thromboembolism in patients with cancer - a cohort study using linked United Kingdom databases. , 2013, European journal of cancer.

[22]  J. Ben-Ezra,et al.  A proposed role for neutrophil extracellular traps in cancer immunoediting , 2013, Front. Immunol..

[23]  M. Dikshit,et al.  Cytokines Induced Neutrophil Extracellular Traps Formation: Implication for the Inflammatory Disease Condition , 2012, PloS one.

[24]  P. Kubes,et al.  Intravascular neutrophil extracellular traps capture bacteria from the bloodstream during sepsis. , 2012, Cell host & microbe.

[25]  D. Scadden,et al.  Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-associated thrombosis , 2012, Proceedings of the National Academy of Sciences.

[26]  Z. Werb,et al.  Platelets induce neutrophil extracellular traps in transfusion-related acute lung injury. , 2012, The Journal of clinical investigation.

[27]  J. West,et al.  Risk of Venous Thromboembolism in Patients with Cancer: A Systematic Review and Meta-Analysis , 2012, PLoS medicine.

[28]  A. Walch,et al.  Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo , 2012, The Journal of experimental medicine.

[29]  K. Preissner,et al.  Neutrophil Extracellular Traps Directly Induce Epithelial and Endothelial Cell Death: A Predominant Role of Histones , 2012, PloS one.

[30]  P. Reitsma,et al.  The relationship between tissue factor and cancer progression: insights from bench and bedside. , 2012, Blood.

[31]  Werner Solbach,et al.  The Impact of Various Reactive Oxygen Species on the Formation of Neutrophil Extracellular Traps , 2012, Mediators of inflammation.

[32]  D. Tuveson,et al.  Understanding Metastasis in Pancreatic Cancer: A Call for New Clinical Approaches , 2012, Cell.

[33]  D. Wagner,et al.  Neutrophil extracellular traps promote deep vein thrombosis in mice , 2012, Journal of thrombosis and haemostasis : JTH.

[34]  Robert A. Campbell,et al.  Novel Anti-bacterial Activities of β-defensin 1 in Human Platelets: Suppression of Pathogen Growth and Signaling of Neutrophil Extracellular Trap Formation , 2011, PLoS pathogens.

[35]  J. Michel,et al.  Early Atheroma-Derived Agonists of Peroxisome Proliferator–Activated Receptor-&ggr; Trigger Intramedial Angiogenesis in a Smooth Muscle Cell–Dependent Manner , 2011, Circulation research.

[36]  M. Glogauer,et al.  Rac2 is required for the formation of neutrophil extracellular traps , 2011, Journal of leukocyte biology.

[37]  D. Wagner,et al.  Histones induce rapid and profound thrombocytopenia in mice. , 2011, Blood.

[38]  T. Wurdinger,et al.  Blood platelets contain tumor-derived RNA biomarkers. , 2011, Blood.

[39]  C. Esmon,et al.  Extracellular histones promote thrombin generation through platelet-dependent mechanisms: involvement of platelet TLR2 and TLR4. , 2011, Blood.

[40]  D. Mottier,et al.  Venous thromboembolism in patients with pancreatic cancer: implications of circulating tissue factor , 2011, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[41]  Laurie J. Gay,et al.  Contribution of platelets to tumour metastasis , 2011, Nature Reviews Cancer.

[42]  M. Surette,et al.  A Novel Mechanism of Rapid Nuclear Neutrophil Extracellular Trap Formation in Response to Staphylococcus aureus , 2010, The Journal of Immunology.

[43]  H. Sørensen,et al.  Hospitalisation for venous thromboembolism in cancer patients and the general population: a population-based cohort study in Denmark, 1997–2006 , 2010, British Journal of Cancer.

[44]  J. Hartwig,et al.  Extracellular DNA traps promote thrombosis , 2010, Proceedings of the National Academy of Sciences.

[45]  Abdul Hakkim,et al.  Impairment of neutrophil extracellular trap degradation is associated with lupus nephritis , 2010, Proceedings of the National Academy of Sciences.

[46]  W. Nacken,et al.  Neutrophil Extracellular Traps Contain Calprotectin, a Cytosolic Protein Complex Involved in Host Defense against Candida albicans , 2009, PLoS pathogens.

[47]  P. Lin,et al.  Chemical and molecular mechanisms of antioxidants: experimental approaches and model systems , 2009, Journal of cellular and molecular medicine.

[48]  G. Cheng,et al.  Polarization of tumor-associated neutrophil phenotype by TGF-beta: "N1" versus "N2" TAN. , 2009, Cancer cell.

[49]  T. Yip,et al.  Platelets actively sequester angiogenesis regulators. , 2009, Blood.

[50]  T. Wilson,et al.  Cathepsin G enhances mammary tumor-induced osteolysis by generating soluble receptor activator of nuclear factor-kappaB ligand. , 2008, Cancer research.

[51]  Stephen R. Clark,et al.  Platelet TLR4 activates neutrophil extracellular traps to ensnare bacteria in septic blood , 2007, Nature Medicine.

[52]  V. Wahn,et al.  Novel cell death program leads to neutrophil extracellular traps. , 2007, The Journal of cell biology.

[53]  D. Zurakowski,et al.  Use of a collagen‐platelet rich plasma scaffold to stimulate healing of a central defect in the canine ACL , 2006, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[54]  W. Holzgreve,et al.  Induction of neutrophil extracellular DNA lattices by placental microparticles and IL-8 and their presence in preeclampsia. , 2005, Human immunology.

[55]  P. Jurasz,et al.  Platelet–cancer interactions: mechanisms and pharmacology of tumour cell‐induced platelet aggregation , 2004, British journal of pharmacology.

[56]  A. Khorana,et al.  Pancreatic cancer and thromboembolic disease. , 2004, The Lancet. Oncology.

[57]  A. Zychlinsky,et al.  Neutrophil Extracellular Traps Kill Bacteria , 2004, Science.

[58]  M. Levine,et al.  Venous Thromboembolism and Cancer: Risks and Outcomes , 2003, Circulation.

[59]  H. Asakura,et al.  Activated Platelets in Ulcerative Colitis Enhance the Production of Reactive Oxygen Species by Polymorphonuclear Leukocytes , 2001, Scandinavian journal of gastroenterology.

[60]  D. Wagner,et al.  PHAGOCYTES , GRANULOCYTES , AND MYELOPOIESIS P-selectin promotes neutrophil extracellular trap formation in mice , 2015 .