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Tumor shedding and metastatic progression after tumor excision in patient-derived orthotopic xenograft models of triple-negative breast cancer

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Abstract

Patient-derived orthotopic xenograft (PDOX) models have been verified as a useful method for studying human cancers in mice. Previous studies on the extent of metastases in these models have been limited by the necessity of welfare euthanasia (primary tumors reaching threshold size), at which point metastases may only be micrometers in diameter, few in number, and solely identified by step-sectioning of formalin-fixed paraffin-embedded tissue. These small micro-metastases are less suitable for many downstream molecular analyses than macro-metastases. Resection of the primary tumor by survival surgery has been proven to allow further time for metastases to grow. Although PDOX models of triple-negative breast cancer (TNBC) shed circulating tumor cells (CTCs) into the bloodstream and metastasize, similar to human TNBC, little data has been collected in these TNBC PDOX models regarding the association between CTC characteristics and distant metastasis following excision of the primary tumor xenograft. This study assembles a timeline of PDOX tumor shedding and metastatic tumor progression before and after tumor excision surgery. We report the ability to use tumorectomies to increase the lifespan of TNBC PDOX models with the potential to obtain larger metastases. CTC clusters and CTCs expressing a mesenchymal marker (vimentin) were associated with metastatic burden in lung and liver. The data collected through these experiments will guide the further use of PDOX models in studying metastatic TNBC.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on request.

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Acknowledgements

The authors would like to thank the Department of Comparative Medicine, the Master of Laboratory Animal Science (MLAS) Training Program, and the Veterinary Service Center (VSC) at Stanford University for their support of this project. We would especially like to acknowledge and thank Elias Godoy of the VSC for his assistance. This project was funded through the Stanford MLAS Graduate Student Fund (AMR), the John and Marva Warnock Research Fund (SSJ), and NIH grants 4P30CA124435 that supports the Biostatistics Core of the Stanford Cancer Institute (AM) and SPECTRUM award number 1UL1TR003142 (AM).

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Authors and Affiliations

  1. Department of Comparative Medicine, Stanford University School of Medicine, Stanford, CA, USA

    Aryana M. Razmara, Sam W. Baker & Kerriann M. Casey

  2. Vortex Biosciences Inc., Pleasanton, CA, USA

    Elodie Sollier & Clementine A. Lemaire

  3. Department of Surgery, Stanford University School of Medicine, Stanford, CA, USA

    Grace N. Kisirkoi, Vishnu C. Ramani & Stefanie S. Jeffrey

  4. Stanford University, Stanford, CA, USA

    Margot B. Bellon

  5. Department of Biomedical Data Science, Stanford University School of Medicine, Stanford, CA, USA

    Alex McMillan

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  1. Aryana M. Razmara
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Contributions

AMR, ES, GNK, SWB, VCR, SSJ, and KMC designed and planned the experiments. AMR, GNK, SWB, MBB, CAL, and VCR performed the experiments; AMR and KMC performed histologic studies. AMR, AM, SSJ, and KMC analyzed the data. AMR wrote the manuscript with assistance from the other authors. All authors have reviewed and approved this manuscript.

Corresponding authors

Correspondence to Stefanie S. Jeffrey or Kerriann M. Casey.

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Conflict of interests

CAL and ES have financial interests in Vortex Biosciences and intellectual property described herein. SSJ serves as an expert advisor for Ravel Biotechnology, which is developing an analytic platform for early cancer detection using cell-free DNA. All other authors declare no competing interests.

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Razmara, A.M., Sollier, E., Kisirkoi, G.N. et al. Tumor shedding and metastatic progression after tumor excision in patient-derived orthotopic xenograft models of triple-negative breast cancer. Clin Exp Metastasis 37, 413–424 (2020). https://doi.org/10.1007/s10585-020-10033-3

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