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A simple quantitative PCR assay to determine TRAMP transgene zygosity

Prostate Cancer and Prostatic Diseases volume 24, pages 358–361 (2021)Cite this article

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Abstract

Background

The TRansgenic Adenocarcinoma of the Mouse Prostate (TRAMP) model remains one of the most widely used transgenic mouse models of prostate cancer. This is due to its ability to recapitulate with ~100% penetrance multiple aspects of the human disease such as prostatic intraepithelial neoplasia lesions, invasive carcinoma, progression to castration-resistant prostate cancer including aggressive neuroendocrine prostate cancer and metastasis. Despite its popularity, the use of TRAMP mice is limited/slowed by the inability to distinguish the zygosity of the TRAMP transgene. This is especially true for breeding strategies implementing multiple crosses and alleles and when the rapid generation of large animal cohorts with the desired genotype is needed.

Methods

We developed a quantitative PCR (qPCR) approach to determine the relative TRAMP transgene copy number of mice.

Results

This method was validated by three independent laboratories across two institutions, which successfully identified the genotype of the mice 98.2% of the time (165/168) in the first attempt. The genotypes of the uncertain mice were correctly identified in the repeated experiments.

Conclusions

We develop the first straightforward, qPCR approach to reliably determine the TRAMP transgene zygosity. The development of this qPCR-based genotyping method enables researchers to streamline breeding strategies when creating complex genetic mouse models involving TRAMP mice; thus, ultimately reducing the required animal numbers, cost, and investigator time.

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Fig. 1: TRAMP transgene zygosity determined by qPCR analysis.

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References

  1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin. 2019;69:7–34.

    Article  Google Scholar 

  2. Wang G, Zhao D, Spring DJ, DePinho RA. Genetics and biology of prostate cancer. Genes Dev. 2018;32:1105–40.

    Article  CAS  Google Scholar 

  3. Ittmann M, Huang J, Radaelli E, Martin P, Signoretti S, Sullivan R, et al. Animal models of human prostate cancer: the consensus report of the New York meeting of the Mouse Models of Human Cancers Consortium Prostate Pathology Committee. Cancer Res. 2013;73:2718–36.

    Article  CAS  Google Scholar 

  4. Gingrich JR, Barrios RJ, Kattan MW, Nahm HS, Finegold MJ, Greenberg NM. Androgen-independent prostate cancer progression in the TRAMP model. Cancer Res. 1997;57:4687–91.

    CAS  PubMed  Google Scholar 

  5. Gingrich JR, Barrios RJ, Morton RA, Boyce BF, DeMayo FJ, Finegold MJ, et al. Metastatic prostate cancer in a transgenic mouse. Cancer Res. 1996;56:4096–102.

    CAS  PubMed  Google Scholar 

  6. Greenberg NM, DeMayo F, Finegold MJ, Medina D, Tilley WD, Aspinall JO, et al. Prostate cancer in a transgenic mouse. Proc Natl Acad Sci USA. 1995;92:3439–43.

    Article  CAS  Google Scholar 

  7. Chiaverotti T, Couto SS, Donjacour A, Mao JH, Nagase H, Cardiff RD, et al. Dissociation of epithelial and neuroendocrine carcinoma lineages in the transgenic adenocarcinoma of mouse prostate model of prostate cancer. Am J Pathol. 2008;172:236–46.

    Article  CAS  Google Scholar 

  8. Hurwitz AA, Foster BA, Allison JP, Greenberg NM, Kwon ED. The TRAMP mouse as a model for prostate cancer. Curr Protoc Immunol. 2001;Chapter 20:Unit 20 5.

    PubMed  Google Scholar 

  9. Berman-Booty LD, Knudsen KE. Models of neuroendocrine prostate cancer. Endocr Relat Cancer. 2015;22:R33–49.

    Article  CAS  Google Scholar 

  10. Gelman IH. How the TRAMP model revolutionized the study of prostate cancer progression. Cancer Res. 2016;76:6137–9.

    Article  CAS  Google Scholar 

  11. Grabowska MM, Matusik RJ. Therapy-induced small-cell disease: from mouse to man and back. Nat Rev Urol. 2018;15:662–3.

    Article  Google Scholar 

  12. Aggarwal R, Huang J, Alumkal JJ, Zhang L, Feng FY, Thomas GV, et al. Clinical and genomic characterization of treatment-emergent small-cell neuroendocrine prostate cancer: a multi-institutional prospective study. J Clin Oncol. 2018;36:2492–503.

    Article  CAS  Google Scholar 

  13. Davies AH, Beltran H, Zoubeidi A. Cellular plasticity and the neuroendocrine phenotype in prostate cancer. Nat Rev Urol. 2018;15:271–86.

    Article  CAS  Google Scholar 

  14. Aparicio AM, Shen L, Tapia EL, Lu JF, Chen HC, Zhang J, et al. Combined tumor suppressor defects characterize clinically defined aggressive variant prostate cancers. Clin Cancer Res. 2016;22:1520–30.

    Article  CAS  Google Scholar 

  15. Beltran H, Prandi D, Mosquera JM, Benelli M, Puca L, Cyrta J, et al. Divergent clonal evolution of castration-resistant neuroendocrine prostate cancer. Nat Med. 2016;22:298–305.

    Article  CAS  Google Scholar 

  16. Beltran H, Rickman DS, Park K, Chae SS, Sboner A, MacDonald TY, et al. Molecular characterization of neuroendocrine prostate cancer and identification of new drug targets. Cancer Discov. 2011;1:487–95.

    Article  CAS  Google Scholar 

  17. Mu P, Zhang Z, Benelli M, Karthaus WR, Hoover E, Chen CC, et al. SOX2 promotes lineage plasticity and antiandrogen resistance in TP53- and RB1-deficient prostate cancer. Science. 2017;355:84–8.

    Article  CAS  Google Scholar 

  18. Ku SY, Rosario S, Wang Y, Mu P, Seshadri M, Goodrich ZW, et al. Rb1 and Trp53 cooperate to suppress prostate cancer lineage plasticity, metastasis, and antiandrogen resistance. Science. 2017;355:78–83.

    Article  CAS  Google Scholar 

  19. Zou M, Toivanen R, Mitrofanova A, Floch N, Hayati S, Sun Y, et al. Transdifferentiation as a mechanism of treatment resistance in a mouse model of castration-resistant prostate cancer. Cancer Discov. 2017;7:736–49.

    Article  CAS  Google Scholar 

  20. Lee DK, Liu Y, Liao L, Li W, Danielpour D, Xu J. Neuroendocrine prostate carcinoma cells originate from the p63-expressing basal cells but not the pre-existing adenocarcinoma cells in mice. Cell Res. 2019;29:420–2.

    Article  Google Scholar 

  21. Ding Z, Wu CJ, Chu GC, Xiao Y, Ho D, Zhang J, et al. SMAD4-dependent barrier constrains prostate cancer growth and metastatic progression. Nature. 2011;470:269–73.

    Article  CAS  Google Scholar 

  22. Truett GE, Heeger P, Mynatt RL, Truett AA, Walker JA, Warman ML. Preparation of PCR-quality mouse genomic DNA with hot sodium hydroxide and tris (HotSHOT). Biotechniques. 2000;29:52–4.

    Article  CAS  Google Scholar 

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Acknowledgements

GW and DEF are supported by the Prostate Cancer Moon Shot and Institutional Research Grant (IRG) Programs at the University of Texas MD Anderson Cancer Center. GW and DEF are also supported by the National Institutes of Health (NIH) Prostate Cancer SPORE (P50 CA140388). GW is additionally supported by a University of Texas Star Award and NIH grant R00CA194289. DEF is further supported by NIH grant R01CA184208 and a grant from the American Cancer Society (RSG-16-084-01 — TBE). BAF is supported by National Cancer Institute (NCI) grant P30CA016056 involving the use of Roswell Park Comprehensive Cancer Center’s Genomic Shared Resource.

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Author notes

  1. These authors contributed equally: Ruidong Chen, Xin Liang

Authors and Affiliations

  1. Department of Genitourinary Medical Oncology and the David H. Koch Center for Applied Research of Genitourinary Cancers, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Ruidong Chen, Xin Liang, Ming Zhu, Daniel E. Frigo & Guocan Wang

  2. Department of Cancer Systems Imaging, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Mollianne M. Murray, Jenny J. Han & Daniel E. Frigo

  3. Department of Pharmacology and Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, NY, USA

    Ellen Karasik & Barbara A. Foster

  4. Center for Nuclear Receptors and Cell Signaling, Department of Biology and Biochemistry, University of Houston, Houston, TX, USA

    Daniel E. Frigo

  5. The Houston Methodist Research Institute, Houston, TX, USA

    Daniel E. Frigo

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  1. Ruidong Chen
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  2. Xin Liang
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  3. Mollianne M. Murray
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  7. Barbara A. Foster
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Corresponding authors

Correspondence to Barbara A. Foster, Daniel E. Frigo or Guocan Wang.

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Conflict of interest

DEF has received research funding from GTx, Inc and has a familial relationship with Maia Biotechnology, Alms Therapeutics, Hinova Pharmaceuticals and Barricade Therapeutics. The other authors report no potential conflicts of interest.

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Chen, R., Liang, X., Murray, M.M. et al. A simple quantitative PCR assay to determine TRAMP transgene zygosity. Prostate Cancer Prostatic Dis 24, 358–361 (2021). https://doi.org/10.1038/s41391-020-00282-4

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