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Orthotopic Implantation Achieves Better Engraftment and Faster Growth Than Subcutaneous Implantation in Breast Cancer Patient-Derived Xenografts

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Abstract

Patient-Derived Xenograft (PDX) is now accepted as a murine model that better mimics human cancer when compared to a conventional cancer cell-line inoculation model. Some claim the advantage of orthotopic site implantation of patient tumor (OS) over ectopic implantation into the subcutaneous space (SQ); however, there has been no study that describes a head-to-head comparison of oncological differences between these two models to date. We hypothesize that OS tumors re-transplant and grow better than SQ tumors and are therefore a better model to evaluate tumor aggressiveness. Breast cancer PDXs were generated using the tumors derived from 11 patients into NOD scid gamma (NSG) mice. We used six ER(+)HER2(−) tumors and five triple negative (TN) tumors for a total of 11 tumors. Five PDX lines grew for an overall engraftment rate of 45%. We present our OS implantation method in detail. The re-transplantation rate of TN tumors in each transplant site was significantly higher in OS when compared to SQ tumors (70.1% vs. 32.1%, p < 0.01). OS tumors grow significantly faster than SQ tumors. Similarly, OS tumors demonstrated significantly more mitotic figures and Ki-67 positive cells than SQ tumors. The tumor re-transplantation rate significantly increased by the second and third generations with the OS method. The time from implantation to development of a palpable tumor dramatically decreased after the first passage. PDX of ER(+) tumors demonstrated significantly lower engraftment rates and slower tumor growth than TN tumors, which remarkably improved by the first passage. Orthotopically implanted PDX tumors showed better re-transplantation rates, greater tumor size, and more significant growth compared to the subcutaneously implanted model.

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Abbreviations

BC:

Breast Cancer

PDX:

Patient-Derived Xenografts

MFP:

Mammary Fat Pads

OS:

Orthotopic Site

SQ:

Subcutaneous

TNBC:

Triple-negative breast cancer

ER(+):

Estrogen receptor positive

NSG:

NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ

HER2:

Human epidermal growth factor receptor 2

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Acknowledgements

This work was supported by NIH grant R01CA160688 to K.T, and National Cancer Institute (NCI) grant P30CA016056 involving the use of Roswell Park Comprehensive Cancer Center’s Shared Resources.

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Authors and Affiliations

  1. Breast Surgery, Department of Surgical Oncology, Roswell Park Comprehensive Cancer Center, Elm & Carlton Streets, Buffalo, NY, 14263, USA

    Maiko Okano, Masanori Oshi, Ali Butash, Ichiro Okano, Tsutomu Kawaguchi & Kazuaki Takabe

  2. Department of Breast Surgery, Fukushima Medical University School of Medicine, Fukushima, Japan

    Maiko Okano, Toru Ohtake & Kazuaki Takabe

  3. Department of Gastrointestinal Tract Surgery, Fukushima Medical University School of Medicine, Fukushima, Japan

    Katsuharu Saito & Koji Kono

  4. Division of Digestive and General Surgery, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan

    Masayuki Nagahashi & Kazuaki Takabe

  5. Department of Surgery, University at Buffalo Jacobs School of Medicine and Biomedical Sciences, The State University of New York, Buffalo, NY, USA

    Kazuaki Takabe

  6. Department of Breast Surgery and Oncology, Tokyo Medical University, Tokyo, Japan

    Kazuaki Takabe

  7. Department of Surgery, Yokohama City University, Yokohama, Japan

    Kazuaki Takabe

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  1. Maiko Okano
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  2. Masanori Oshi
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Contributions

M.O. conceptualized study design, performed experiments and prepared the article.

M.O., I.O., K.S. and T.K. performed experiments.

A.B. prepared the article.

I.O. and M.N. prepared the figs.

K.K. and T.O. secured the funding.

K.T. provided supervision of experiments and prepared the article.

Corresponding author

Correspondence to Kazuaki Takabe.

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Supplementary figure 1.

Comparisons of representative histology of patients’ tumor and Xenografts. Hematoxilyn-Eosin (H&E) staining and Immunohistochemical staining of ANXA1, ER, PgR and HER2 in each tumors. (PNG 7092 kb)

High resolution image (TIF 1517 kb)

Supplementary figure 2.

Representative growth curves of other individual tumors. SQ (solid line), OS (broken line) tumors. (PNG 834 kb)

High resolution image (TIF 118 kb)

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Okano, M., Oshi, M., Butash, A. et al. Orthotopic Implantation Achieves Better Engraftment and Faster Growth Than Subcutaneous Implantation in Breast Cancer Patient-Derived Xenografts. J Mammary Gland Biol Neoplasia 25, 27–36 (2020). https://doi.org/10.1007/s10911-020-09442-7

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