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Establishing a human renal cell carcinoma tumorgraft platform for preclinical drug testing

Nature Protocols volume 9, pages 1848–1859 (2014)Cite this article

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Abstract

Traditionally, xenograft models have been used to study tumors in vivo. However, their utility is reduced by the use of tumor cell lines for implantation. Tumorgrafts (TGs; also known as patient-derived xenografts (PDXs)), which involve patient-derived tumor samples, are increasingly recognized as more representative models than traditional xenografts. Furthermore, we showed previously that renal cell carcinoma (RCC) TGs retain the histology, gene expression, DNA copy number alterations, mutations and treatment responsiveness of patient tumors. In skilled hands, implantations require ≤5 min per mouse, and TGs typically grow to 1 cm in 1–4 months. Here we outline the process of implantation of patient-derived RCC samples into the kidneys of immunodeficient mice, as well as the s.c. implantation for preclinical drug testing, including guidelines for the design and execution of drug trials. TGs have extensive applications besides therapeutic studies and may identify biomarkers and mechanisms of resistance. In addition, they may provide insights into tumor biology.

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Figure 1: Chromatogram showing VHL (encoding von Hippel-Lindau tumor suppressor, E3 ubiquitin protein ligase) mutant allele enrichment in TGs.
Figure 2: Matched patient (left) and TG tumors (right) for three different clear cell RCC cases.
Figure 3: Identifying kidney tumors under the microscope.
Figure 4: Renal capsule implantation.
Figure 5: Harvesting tissue from a tumor-bearing mouse.
Figure 6: Subcutaneous implantation for drug trials.
Figure 7: Subcutaneous tumor after euthanasia of drug trial mouse.

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Acknowledgements

We are thankful to S. Sivanand for her help in establishing the program, P. Kapur for providing RCC histology pictures, and S. Cohn, R. McKay, S. Peña-Llopis, T. Anh Tran and S. Vega-Rubín-de-Celis for reviewing the manuscript. The work described herein was supported by grants from the American Cancer Society (RSG115739), the V Foundation, the US National Institutes of Health (R01CA175754), and the Cancer Prevention and Research Institute of Texas (RP101075, RP130172 and RP130603) to J.B., as well as a Cancer Center Core grant (P30CA142543). J.B. is a Virginia Murchison Linthicum Endowed Scholar in Medical Research.

Author information

Authors and Affiliations

  1. Kidney Cancer Program, University of Texas (UT) Southwestern Medical Center, Dallas, Texas, USA

    Andrea Pavía-Jiménez, Vanina Toffessi Tcheuyap & James Brugarolas

  2. Department of Internal Medicine, UT Southwestern Medical Center, Dallas, Texas, USA

    Andrea Pavía-Jiménez, Vanina Toffessi Tcheuyap & James Brugarolas

  3. Department of Developmental Biology, UT Southwestern Medical Center, Dallas, Texas, USA

    Andrea Pavía-Jiménez, Vanina Toffessi Tcheuyap & James Brugarolas

  4. Simmons Comprehensive Cancer Center, UT Southwestern Medical Center, Dallas, Texas, USA

    Andrea Pavía-Jiménez, Vanina Toffessi Tcheuyap & James Brugarolas

Authors
  1. Andrea Pavía-Jiménez
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  2. Vanina Toffessi Tcheuyap
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  3. James Brugarolas
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Contributions

A.P.-J. wrote the manuscript with input from other authors. V.T.T. contributed to the first draft of the manuscript. J.B. conceived and orchestrated the setup of the tumorgraft platform with assistance from A.P.-J., V.T.T. and other laboratory members, and contributed to the writing of the manuscript.

Corresponding author

Correspondence to James Brugarolas.

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Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Identifying mice with Thymic Lymphoma (TL).

a) Mice with TL are less active, short of breath, and may have bulging eyes. b) Healthy mouse for comparison. c) Microscopic infiltration of TL cells (small cells with little to no cytoplasm) in a normal NOD/SCID kidney. Clusters of lymphoma cells are outlined in black. d) Carcass of mouse with TL showing large white mass in the chest cavity. The heart and lungs can be found behind this mass. The spleen may also be enlarged (not shown). All mouse experiments were approved by UT Southwestern Medical Center's IACUC.

Supplementary Figure 2 Hood setup.

From left to right: (Back row) hot bead sterilizer, tissue to be implanted in petri dish, induction chamber, 70% alcohol spray, surgery board with heating pad, isoflurane vaporizer. (Front row) Ear tags and ear tag applicator, 'external' tools, 'internal' tools.

Supplementary information

Supplementary Figure 1

Identifying mice with Thymic Lymphoma (TL). (PDF 8347 kb)

Supplementary Figure 2

Hood setup. (PDF 180 kb)

Supplementary Table 1

Database tabs to facilitate colony maintenance. (XLSX 16 kb)

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Pavía-Jiménez, A., Tcheuyap, V. & Brugarolas, J. Establishing a human renal cell carcinoma tumorgraft platform for preclinical drug testing. Nat Protoc 9, 1848–1859 (2014). https://doi.org/10.1038/nprot.2014.108

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