title: Targeting cMET with INC280 impairs tumour growth and improves efficacy of gemcitabine in a pancreatic cancer model.
abstract: BACKGROUND
  Expression and activation of the cMET receptor have been implicated in tumor progression and resistance to chemotherapy in human pancreatic cancer. In this regard we assessed the effects of targeting cMET in pancreatic cancer models in vitro and in vivo.
  
  
  METHODS
  Human (L3.6pl, BxP3, HPAF-II, MiaPaCa2) and murine (Panc02) pancreatic cancer cell lines, endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) were used for the experiments. Furthermore, the human pancreatic cancer cell line MiaPaCa2 with acquired resistance to gemcitabine was employed (MiaPaCa2(G250)). For targeting the cMET receptor, the oral available, ATP-competitive inhibitor INC280 was used. Effects of cMET inhibition on cancer and stromal cells were determined by growth assays, western blotting, motility assays and ELISA. Moreover, orthotopic xenogeneic and syngeneic mouse (BALB-C nu/nu; C57BL/6) models were used to assess in vivo efficacy of targeting cMET alone and in combination with gemcitabine.
  
  
  RESULTS
  Treatment with INC280 impairs activation of signaling intermediates in pancreatic cancer cells and ECs, particularly when cells were stimulated with hepatocyte growth factor (HGF). Moreover, motility of cancer cells and ECs in response to HGF was reduced upon treatment with INC280. Only minor effects on VSMCs were detected. Interestingly, MiaPaCa2(G250) showed an increase in cMET expression and cMET inhibition abrogated HGF-induced effects on growth, motility and signaling as well as DFX-hypoxia HIF-1alpha and MDR-1 expression in vitro. In vivo, therapy with INC280 alone led to inhibition of orthotopic tumor growth in xenogeneic and syngeneic models. Similar to in vitro results, cMET expression was increased upon treatment with gemcitabine, and combination of the cMET inhibitor with gemcitabine improved anti-neoplastic capacity in an orthotopic syngeneic model. Immunohistochemical analysis revealed a significant inhibition of tumor cell proliferation (Ki67) and tumor vascularization (CD31). Finally, combination of gemcitabine with INC280 significantly prolonged survival in the orthotopic syngeneic tumor model even when treatment with the cMET inhibitor was initiated at an advanced stage of disease.
  
  
  CONCLUSIONS
  These data provide evidence that targeting cMET in combination with gemcitabine may be effective in human pancreatic cancer and warrants further clinical evaluation.
authors.affiliation: Department of Surgery, University Hospital Regensburg, University of Regensburg, Franz-Josef-Strauss Allee 11, 93053, Regensburg, Germany. franziska.brandes@gmail.com.
authors.forename: Franziska
authors.initials: F
authors.lastname: Brandes
authors.affiliation: Department of Surgery, University Hospital Regensburg, University of Regensburg, Franz-Josef-Strauss Allee 11, 93053, Regensburg, Germany. katharina.schmidt@ukr.de.
authors.forename: Katharina
authors.initials: K
authors.lastname: Schmidt
authors.affiliation: Department of Surgery, University Hospital Regensburg, University of Regensburg, Franz-Josef-Strauss Allee 11, 93053, Regensburg, Germany. christine.wagner@ukr.de.
authors.forename: Christine
authors.initials: C
authors.lastname: Wagner
authors.affiliation: Department of Surgery, University Hospital Regensburg, University of Regensburg, Franz-Josef-Strauss Allee 11, 93053, Regensburg, Germany. julia.redekopf@ukr.de.
authors.forename: Julia
authors.initials: J
authors.lastname: Redekopf
authors.affiliation: Department of Surgery, University Hospital Regensburg, University of Regensburg, Franz-Josef-Strauss Allee 11, 93053, Regensburg, Germany. hans.schlitt@ukr.de.
authors.forename: Hans Jürgen
authors.initials: HJ
authors.lastname: Schlitt
authors.affiliation: Department of Surgery, University Hospital Regensburg, University of Regensburg, Franz-Josef-Strauss Allee 11, 93053, Regensburg, Germany. edward.geissler@ukr.de.
authors.forename: Edward Kenneth
authors.initials: EK
authors.lastname: Geissler
authors.affiliation: Department of Surgery, University Hospital Regensburg, University of Regensburg, Franz-Josef-Strauss Allee 11, 93053, Regensburg, Germany. sven.lang@ukr.de.
authors.forename: Sven Arke
authors.initials: SA
authors.lastname: Lang
mesh_terms: D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000008:administration & dosage* / Q000627:therapeutic use; D001549:Benzamides / Q000008:administration & dosage* / Q000494:pharmacology; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000008:administration & dosage* / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003841:Deoxycytidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000627:therapeutic use; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D004357:Drug Synergism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007093:Imidazoles; D051379:Mice; D058990:Molecular Targeted Therapy; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D019859:Proto-Oncogene Proteins c-met / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014227:Triazines; D023041:Xenograft Model Antitumor Assays; D000093542:Gemcitabine
chemical_list: D000964:Antimetabolites, Antineoplastic; D001549:Benzamides; D019086:Bridged Bicyclo Compounds, Heterocyclic; D007093:Imidazoles; D014227:Triazines; D003841:Deoxycytidine; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met; C000613976:capmatinib; D000093542:Gemcitabine