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

Abstract

BackgroundThe 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.MethodsWe 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.ResultsWe 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.ConclusionsOur 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-015-0469-1) contains supplementary material, which is available to authorized users.

Stromal SPOCK1 supports invasive pancreatic cancer growth

“…For detailed description of primary cell line isolation, propagation, and characterization, see Damhofer et al . ().…”

“…All surgical procedures were performed under isoflurane anesthesia. For detailed description of primary cell line isolation, propagation, and characterization, see Damhofer et al (2015).…”

“…Tissue slides were stained as previously described on the paraffin-embedded slides (Damhofer et al, 2015). Antibodies and dilutions used were CK19 1 : 500 (MU246-UC; Biogenex), CXCR4 1 : 400 (Ab124824; Abcam, Cambridge, UK), Ki67 1 : 2000 (SAB5500134; Sigma).…”

Preclinical models for interrogating drug action in human cancers using Stable Isotope Resolved Metabolomics (SIRM)

“…Even when some case studies have shown that the PDX model can predict the human drug response (Whittle et al 2015; Maykel J. et al 2014), the fact remains that the mouse model may not recapitulate the off target effects as there is no corresponding implanted normal human tissues. Finally, the long duration for establishing resected human tumors in PDX model and in some instances the low “take rate” compromises the need for timely feedback on drug testing to inform on efficacious personalized treatment (Hidalgo et al 2011) (Damhofer et al 2015; Aparicio et al 2015). …”

MiR-329 restricts tumor growth by targeting GRB2 in pancreatic cancer

“…Cells were maintained in DMEM supplemented with 10% FBS (GIBCO) at 37 o C under 5% CO 2 in a humidified chamber. Primary pancreatic cancer cells were cultured as Helene et al [32] described and maintained in RPMI 1640 with 10% FBS.…”