title: Establishment of patient-derived xenograft models and cell lines for malignancies of the upper gastrointestinal tract.
abstract: BACKGROUND
  The upper gastrointestinal tract is home to some of most notorious cancers like esophagogastric and pancreatic cancer. Several factors contribute to the lethality of these tumors, but one that stands out for both tumor types is the strong inter- as well as intratumor heterogeneity. Unfortunately, genetic tumor models do not match this heterogeneity, and for esophageal cancer no adequate genetic models exist. To allow for an improved understanding of these diseases, tissue banks with sufficient amount of samples to cover the extent of diversity of human cancers are required. Additionally, xenograft models that faithfully mimic and span the breadth of human disease are essential to perform meaningful functional experiments.
  
  
  METHODS
  We describe here the establishment of a tissue biobank, patient derived xenografts (PDXs) and cell line models of esophagogastric and pancreatic cancer patients. Biopsy material was grafted into immunocompromised mice and PDXs were used to establish primary cell cultures to perform functional studies. Expression of Hedgehog ligands in patient tumor and matching PDX was assessed by immunohistochemical staining, and quantitative real-time PCR as well as flow cytometry was used for cultured cells. Cocultures with Hedgehog reporter cells were performed to study paracrine signaling potency. Furthermore, SHH expression was modulated in primary cultures using lentiviral mediated knockdown.
  
  
  RESULTS
  We have established a panel of 29 PDXs from esophagogastric and pancreatic cancers, and demonstrate that these PDXs mirror several of the (immuno)histological and biochemical characteristics of the original tumors. Derived cell lines can be genetically manipulated and used to further study tumor biology and signaling capacity. In addition, we demonstrate an active (paracrine) Hedgehog signaling mode by both tumor types, the magnitude of which has not been compared directly in previous studies.
  
  
  CONCLUSIONS
  Our established PDXs and their matching primary cell lines retain important characteristics seen in the original tumors, and this should enable future studies to address the responses of these tumors to different treatment modalities, but also help in gaining mechanistic insight in how some tumors respond to certain regimens and others do not.
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. h.damhofer@amc.uva.nl.
authors.forename: Helene
authors.initials: H
authors.lastname: Damhofer
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. e.a.ebbing@amc.uva.nl.
authors.forename: Eva A
authors.initials: EA
authors.lastname: Ebbing
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. a.steins@amc.uva.nl.
authors.forename: Anne
authors.initials: A
authors.lastname: Steins
authors.affiliation: Department of Surgery, Leiden University Medical Center, Albinusdreef 2, Leiden, ZA, 2333, The Netherlands. L.Welling@lumc.nl.
authors.forename: Lieke
authors.initials: L
authors.lastname: Welling
authors.affiliation: Department of Surgery, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. j.tol@amc.uva.nl.
authors.forename: Johanna A
authors.initials: JA
authors.lastname: Tol
authors.affiliation: Department of Gastroenterology and Hepatology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. k.k.krishnadath@amc.uva.nl.
authors.forename: Kausilia K
authors.initials: KK
authors.lastname: Krishnadath
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. t.vanleusden@amc.uva.nl.
authors.forename: Tom
authors.initials: T
authors.lastname: van Leusden
authors.affiliation: Department of Pathology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. m.j.vandevijver@amc.uva.nl.
authors.forename: Marc J
authors.initials: MJ
authors.lastname: van de Vijver
authors.affiliation: Department of Surgery, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. m.g.besselink@amc.uva.nl.
authors.forename: Marc G
authors.initials: MG
authors.lastname: Besselink
authors.affiliation: Department of Surgery, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. o.r.busch@amc.uva.nl.
authors.forename: Olivier R
authors.initials: OR
authors.lastname: Busch
authors.affiliation: Department of Surgery, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. m.i.vanbergehenegouwen@amc.uva.nl.
authors.forename: Mark I
authors.initials: MI
authors.lastname: van Berge Henegouwen
authors.affiliation: Department of Radiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. o.m.vandelden@amc.uva.nl.
authors.forename: Otto
authors.initials: O
authors.lastname: van Delden
authors.affiliation: Department of Pathology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. s.l.meijer@amc.uva.nl.
authors.forename: Sybren L
authors.initials: SL
authors.lastname: Meijer
authors.affiliation: Department of Pathology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. f.dijk@amc.uva.nl.
authors.forename: Frederike
authors.initials: F
authors.lastname: Dijk
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. j.p.medema@amc.uva.nl.
authors.forename: Jan Paul
authors.initials: JP
authors.lastname: Medema
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. h.vanlaarhoven@amc.uva.nl.|Department of Medical Oncology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. h.vanlaarhoven@amc.uva.nl.
authors.forename: Hanneke W
authors.initials: HW
authors.lastname: van Laarhoven
authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Academic Medical Center, Meibergdreef 9, Amsterdam, AZ, 1105, The Netherlands. m.f.bijlsma@amc.uva.nl.
authors.forename: Maarten F
authors.initials: MF
authors.lastname: Bijlsma
mesh_terms: D000368:Aged; D000818:Animals; D045744:Cell Line, Tumor; D004067:Digestive System Neoplasms / Q000378:metabolism / Q000473:pathology*; D005260:Female; D017930:Genes, Reporter; D053823:Hedgehog Proteins / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008297:Male; D051379:Mice; D008875:Middle Aged; D019899:Paracrine Communication; D015398:Signal Transduction; D017154:Stromal Cells / Q000473:pathology; D014015:Tissue Banks; D041742:Upper Gastrointestinal Tract / Q000378:metabolism / Q000473:pathology*; D023041:Xenograft Model Antitumor Assays
chemical_list: D053823:Hedgehog Proteins; D008024:Ligands