Annals of Surgical Oncology

, Volume 23, Issue 1, pp 305–311 | Cite as

Intramuscular Transplantation Improves Engraftment Rates for Esophageal Patient-Derived Tumor Xenografts

  • Matthew Read
  • David Liu
  • Cuong P. Duong
  • Carleen Cullinane
  • William K. Murray
  • Christina M. Fennell
  • Jake Shortt
  • David Westerman
  • Paul Burton
  • Nicholas J. Clemons
  • Wayne A. Phillips
Translational Research and Biomarkers

Abstract

Background

Recently, there has been an increase in the availability of targeted molecular therapies for cancer treatment. The application of these approaches to esophageal cancer, however, has been hampered by the relative lack of appropriate models for preclinical testing. Patient-derived tumor xenograft (PDTX) models are gaining popularity for studying many cancers. Unfortunately, it has proven difficult to generate xenografts from esophageal cancer using these models. The purpose of this study was to improve the engraftment efficiency of esophageal PDTXs.

Methods

Fresh pieces of esophageal tumors obtained from endoscopic biopsies or resected specimens were collected from 23 patients. The tumors were then coated in Matrigel and transplanted in immunocompromised mice subcutaneously (n = 6) and/or using a novel implantation technique whereby the tumor is placed in a dorsal intramuscular pocket (n = 18). They are then monitored for engraftment.

Results

With the novel intramuscular technique, successful engraftment was achieved for all 18 patient tumors. Among these PDTXs, 13 recapitulated the original patient tumors with respect to degree of differentiation, molecular and genetic profiles, and chemotherapeutic response. Lymphomatous transformation was observed in the other five PDTXs. Successful engraftment was achieved for only one of six patient tumors using the classic subcutaneous approach.

Discussion

We achieved a much higher engraftment rate of PDTXs using our novel intramuscular transplant technique than has been reported in other published studies. It is hoped that this advancement will help expedite the development and testing of new therapies for esophageal cancer.

Supplementary material

10434_2015_4425_MOESM1_ESM.tif (2.9 mb)
Supplementary Fig. 1 Intramuscular (IM) implantation technique and PDTX formation. a After placement of a stay suture, a small IM pocket is created on the dorsum of the mouse that is just large enough to accommodate the tumor piece. b After implantation, the IM pocket is sutured closed with the implant evident immediately below the outer muscle fibers. c Gross view of an IM PDTX at the time of harvest (TIFF 2961 kb)
10434_2015_4425_MOESM2_ESM.tif (3.3 mb)
Supplementary Fig. 2 Identification and characterization of lymphomagenic transformation within a subset of established PDTXs. Immunohistochemistry using pan-cytokeratin AE1/AE3 (1:100) (NCL-L-AE1/AE3; Novocastra/Leica Microsystems, Newcastle, UK), CD20 (1:200) (M7055, DakoCytomation; Dako, Glostrup, Denmark), and CD45 (1:200) (M0701, DakoCytomation) antibodies on the original patient tumor and the corresponding PDTX. The PDTX of patient 9 (a) is negative for the pan-cytokeratin marker AE1/AE3 but positive for the common leukocyte antigen CD45 and the B-lymphocyte restricted antigen CD20, indicating that it is predominantly composed of human B cells. In comparison, the PDTX of patient 16 (b) exhibits a combination of epithelial cells (nests of AE1/AE3-positive cells) and lymphocytes. c Further analysis via flow cytometry was performed as previously described (Craig FE, Foon KA. Blood 2008;222:3941–67) of single cells derived from PDTX 14 and confirmed that the lymphoid cells were CD19+ B cells (left plot). The abnormal B-cell population was then gated and is depicted on the FSC vs. SSC plot in blue (right plot), indicating that these cells are also large. dRight In situ hybridization for the Epstein Barr virus (EBV) (blue staining) was performed on sections of formalin-fixed paraffin-embedded PDTX using the INFORM EBER Probe and ISH iVIEW Blue Detection Kit (Ventana/Roche, Tucson, AZ, USA) as per the manufacturer’s protocol and counterstained with eosin (pink staining). Left EBV-positive nasopharyngeal carcinoma (positive control). Right Section from PDTX 9 (TIFF 3402 kb)

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Copyright information

© Society of Surgical Oncology 2015

Authors and Affiliations

  • Matthew Read
    • 1
    • 2
  • David Liu
    • 1
    • 2
  • Cuong P. Duong
    • 1
    • 3
  • Carleen Cullinane
    • 1
    • 2
  • William K. Murray
    • 5
  • Christina M. Fennell
    • 1
    • 2
  • Jake Shortt
    • 2
    • 6
    • 7
  • David Westerman
    • 5
    • 8
  • Paul Burton
    • 9
    • 10
  • Nicholas J. Clemons
    • 1
    • 2
    • 4
  • Wayne A. Phillips
    • 1
    • 2
    • 3
    • 4
  1. 1.Surgical Oncology Research LaboratoryPeter MacCallum Cancer CentreMelbourneAustralia
  2. 2.Sir Peter MacCallum Department of OncologyUniversity of MelbourneMelbourneAustralia
  3. 3.Division of Cancer SurgeryPeter MacCallum Cancer CentreMelbourneAustralia
  4. 4.Department of Surgery (St. Vincent’s Hospital)University of MelbourneMelbourneAustralia
  5. 5.Department of PathologyPeter MacCallum Cancer CentreMelbourneAustralia
  6. 6.School of Clinical Sciences at Monash HealthMelbourneAustralia
  7. 7.Faculty of Medicine, Nursing & Health SciencesMonash UniversityMelbourneAustralia
  8. 8.University of MelbourneMelbourneAustralia
  9. 9.Monash University Centre for Obesity Research and EducationAlfred HospitalMelbourneAustralia
  10. 10.Cabrini HospitalMelbourneAustralia

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