Paulo L Pfitzinger1, Laura Fangmann1, Kun Wang2, Elke Demir1, Engin Gürlevik3, Bettina Fleischmann-Mundt3, Jennifer Brooks3, Jan G D'Haese4, Steffen Teller1, Andreas Hecker5, Moritz Jesinghaus6, Carsten Jäger1, Lei Ren1,7, Rouzanna Istvanffy1, Florian Kühnel3, Helmut Friess1,8,9, Güralp Onur Ceyhan10, Ihsan Ekin Demir11,12,13,14. 1. Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Ismaninger Str. 22, 81675, Munich, Germany. 2. Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Hepatic, Biliary & Pancreatic Surgery, Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing, 100710, China. 3. Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Hannover, Germany. 4. Department of General, Visceral, and Transplantation Surgery, Ludwig-Maximilians-University Munich, Munich, Germany. 5. Department of General and Thoracic Surgery, University Hospital of Giessen, Giessen, Germany. 6. Institute of Pathology, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany. 7. Department of General Surgery (Gastrointestinal Surgery), The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan, China. 8. German Cancer Consortium (DKTK), Partner Site Munich, Munich, Germany. 9. CRC 1321 Modelling and Targeting Pancreatic Cancer, Munich, Germany. 10. Department of General Surgery, HPB-Unit, School of Medicine, Acibadem Mehmet Ali Aydinlar University, Istanbul, Turkey. 11. Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Ismaninger Str. 22, 81675, Munich, Germany. ekin.demir@tum.de. 12. German Cancer Consortium (DKTK), Partner Site Munich, Munich, Germany. ekin.demir@tum.de. 13. CRC 1321 Modelling and Targeting Pancreatic Cancer, Munich, Germany. ekin.demir@tum.de. 14. Department of General Surgery, HPB-Unit, School of Medicine, Acibadem Mehmet Ali Aydinlar University, Istanbul, Turkey. ekin.demir@tum.de.
Abstract
BACKGROUND: Nerve-cancer interactions are increasingly recognized to be of paramount importance for the emergence and progression of pancreatic cancer (PCa). Here, we investigated the role of indirect cholinergic activation on PCa progression through inhibition of acetylcholinesterase (AChE) via clinically available AChE-inhibitors, i.e. physostigmine and pyridostigmine. METHODS: We applied immunohistochemistry, immunoblotting, MTT-viability, invasion, flow-cytometric-cell-cycle-assays, phospho-kinase arrays, multiplex ELISA and xenografted mice to assess the impact of AChE inhibition on PCa cell growth and invasiveness, and tumor-associated inflammation. Survival analyses were performed in a novel genetically-induced, surgically-resectable mouse model of PCa under adjuvant treatment with gemcitabine+/-physostigmine/pyridostigmine (n = 30 mice). Human PCa specimens (n = 39) were analyzed for the impact of cancer AChE expression on tumor stage and survival. RESULTS: We discovered a strong expression of AChE in cancer cells of human PCa specimens. Inhibition of this cancer-cell-intrinsic AChE via pyridostigmine and physostigmine, or administration of acetylcholine (ACh), diminished PCa cell viability and invasion in vitro and in vivo via suppression of pERK signaling, and reduced tumor-associated macrophage (TAM) infiltration and serum pro-inflammatory cytokine levels. In the novel genetically-induced, surgically-resectable PCa mouse model, adjuvant co-therapy with AChE blockers had no impact on survival. Accordingly, survival of resected PCa patients did not differ based on tumor AChE expression levels. Patients with higher-stage PCa also exhibited loss of the ACh-synthesizing enzyme, choline-acetyltransferase (ChAT), in their nerves. CONCLUSION: For future clinical trials of PCa, direct cholinergic stimulation of the muscarinic signaling, rather than indirect activation via AChE blockade, may be a more effective strategy.
BACKGROUND: Nerve-cancer interactions are increasingly recognized to be of paramount importance for the emergence and progression of pancreatic cancer (PCa). Here, we investigated the role of indirect cholinergic activation on PCa progression through inhibition of acetylcholinesterase (AChE) via clinically available AChE-inhibitors, i.e. physostigmine and pyridostigmine. METHODS: We applied immunohistochemistry, immunoblotting, MTT-viability, invasion, flow-cytometric-cell-cycle-assays, phospho-kinase arrays, multiplex ELISA and xenografted mice to assess the impact of AChE inhibition on PCa cell growth and invasiveness, and tumor-associated inflammation. Survival analyses were performed in a novel genetically-induced, surgically-resectable mouse model of PCa under adjuvant treatment with gemcitabine+/-physostigmine/pyridostigmine (n = 30 mice). Human PCa specimens (n = 39) were analyzed for the impact of cancerAChE expression on tumor stage and survival. RESULTS: We discovered a strong expression of AChE in cancer cells of human PCa specimens. Inhibition of this cancer-cell-intrinsic AChE via pyridostigmine and physostigmine, or administration of acetylcholine (ACh), diminished PCa cell viability and invasion in vitro and in vivo via suppression of pERK signaling, and reduced tumor-associated macrophage (TAM) infiltration and serum pro-inflammatory cytokine levels. In the novel genetically-induced, surgically-resectable PCa mouse model, adjuvant co-therapy with AChE blockers had no impact on survival. Accordingly, survival of resected PCa patients did not differ based on tumorAChE expression levels. Patients with higher-stage PCa also exhibited loss of the ACh-synthesizing enzyme, choline-acetyltransferase (ChAT), in their nerves. CONCLUSION: For future clinical trials of PCa, direct cholinergic stimulation of the muscarinic signaling, rather than indirect activation via AChE blockade, may be a more effective strategy.
Authors: F J Campoy; C J Vidal; E Muñoz-Delgado; M F Montenegro; J Cabezas-Herrera; S Nieto-Cerón Journal: Chem Biol Interact Date: 2016-04-13 Impact factor: 5.192
Authors: Bernhard W Renz; Takayuki Tanaka; Masaki Sunagawa; Ryota Takahashi; Zhengyu Jiang; Marina Macchini; Zahra Dantes; Giovanni Valenti; Ruth A White; Moritz A Middelhoff; Matthias Ilmer; Paul E Oberstein; Martin K Angele; Huan Deng; Yoku Hayakawa; C Benedikt Westphalen; Jens Werner; Helen Remotti; Maximilian Reichert; Yagnesh H Tailor; Karan Nagar; Richard A Friedman; Alina C Iuga; Kenneth P Olive; Timothy C Wang Journal: Cancer Discov Date: 2018-09-05 Impact factor: 39.397
Authors: Lars Ivo Partecke; André Käding; Dung Nguyen Trung; Stephan Diedrich; Matthias Sendler; Frank Weiss; Jens-Peter Kühn; Julia Mayerle; Katharina Beyer; Wolfram von Bernstorff; Claus-Dieter Heidecke; Wolfram Keßler Journal: Oncotarget Date: 2017-04-04