Establishment of patient-derived xenograft models and cell lines for malignancies of the upper gastrointestinal tract View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2015-12

AUTHORS

Helene Damhofer, Eva A Ebbing, Anne Steins, Lieke Welling, Johanna A Tol, Kausilia K Krishnadath, Tom van Leusden, Marc J van de Vijver, Marc G Besselink, Olivier R Busch, Mark I van Berge Henegouwen, Otto van Delden, Sybren L Meijer, Frederike Dijk, Jan Paul Medema, Hanneke W van Laarhoven, Maarten F Bijlsma

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. More... »

PAGES

115

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/s12967-015-0469-1

DOI

http://dx.doi.org/10.1186/s12967-015-0469-1

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1002610071

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/25884700


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