Ink4a/Arf-/- and HRAS(G12V) transform mouse mammary cells into triple-negative breast cancer containing tumorigenic CD49f- quiescent cells

K. Kai, Takayuki Iwamoto, T. Kobayashi, Y. Arima, Y. Takamoto, N. Ohnishi, C. Bartholomeusz, R. Horii, F. Akiyama, G. N. Hortobagyi, L. Pusztai, H. Saya, N. T. Ueno

Research output: Contribution to journalArticle

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Abstract

Intratumoral heterogeneity within individual breast tumors is a well-known phenomenon that may contribute to drug resistance. This heterogeneity is dependent on several factors, such as types of oncogenic drivers and tumor precursor cells. The purpose of our study was to engineer a mouse mammary tumor model with intratumoral heterogeneity by using defined genetic perturbations. To achieve this, we used mice with knockout (-/-) of Ink4a/Arf, a tumor suppressor locus; these mice are known to be susceptible to non-mammary tumors such as fibrosarcoma. To induce mammary tumors, we retrovirally introduced an oncogene, HRAS(G12V), into Ink4a/Arf-/- mammary cells in vitro, and those cells were inoculated into syngeneic mice mammary fat pads. We observed 100% tumorigenesis. The tumors formed were negative for estrogen receptor, progesterone receptor and HER2. Further, they had pathological features similar to those of human triple-negative breast cancer (TNBC) (for example, pushing borders, central necrosis). The tumors were found to be heterogeneous and included two subpopulations: CD49f- quiescent cells and CD49f + cells. Contrary to our expectation, CD49f- quiescent cells had high tumor-initiating potential and CD49f+ cells had relatively low tumor-initiating potential. Gene expression analysis revealed that CD49f- quiescent cells overexpressed epithelial-to-mesenchymal transition-driving genes, reminiscent of tumor-initiating cells and claudin-low breast cancer. Our animal model with intratumoral heterogeneity, derived from defined genetic perturbations, allows us to test novel molecular targeted drugs in a setting that mimics the intratumoral heterogeneity of human TNBC.

Original languageEnglish
Pages (from-to)440-448
Number of pages9
JournalOncogene
Volume33
Issue number4
DOIs
Publication statusPublished - Jan 23 2014
Externally publishedYes

Fingerprint

Triple Negative Breast Neoplasms
Breast
Neoplasms
Breast Neoplasms
Epithelial-Mesenchymal Transition
Neoplastic Stem Cells
Fibrosarcoma
Progesterone Receptors
Oncogenes
Drug Resistance
Knockout Mice
Estrogen Receptors
Adipose Tissue
Carcinogenesis
Necrosis
Animal Models
Gene Expression

Keywords

  • HRAS(G12V)
  • Ink4a/Arf
  • Intratumoral heterogeneity
  • Triple-negative breast cancer
  • Tumor-initiating cell

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Ink4a/Arf-/- and HRAS(G12V) transform mouse mammary cells into triple-negative breast cancer containing tumorigenic CD49f- quiescent cells. / Kai, K.; Iwamoto, Takayuki; Kobayashi, T.; Arima, Y.; Takamoto, Y.; Ohnishi, N.; Bartholomeusz, C.; Horii, R.; Akiyama, F.; Hortobagyi, G. N.; Pusztai, L.; Saya, H.; Ueno, N. T.

In: Oncogene, Vol. 33, No. 4, 23.01.2014, p. 440-448.

Research output: Contribution to journalArticle

Kai, K, Iwamoto, T, Kobayashi, T, Arima, Y, Takamoto, Y, Ohnishi, N, Bartholomeusz, C, Horii, R, Akiyama, F, Hortobagyi, GN, Pusztai, L, Saya, H & Ueno, NT 2014, 'Ink4a/Arf-/- and HRAS(G12V) transform mouse mammary cells into triple-negative breast cancer containing tumorigenic CD49f- quiescent cells', Oncogene, vol. 33, no. 4, pp. 440-448. https://doi.org/10.1038/onc.2012.609
Kai, K. ; Iwamoto, Takayuki ; Kobayashi, T. ; Arima, Y. ; Takamoto, Y. ; Ohnishi, N. ; Bartholomeusz, C. ; Horii, R. ; Akiyama, F. ; Hortobagyi, G. N. ; Pusztai, L. ; Saya, H. ; Ueno, N. T. / Ink4a/Arf-/- and HRAS(G12V) transform mouse mammary cells into triple-negative breast cancer containing tumorigenic CD49f- quiescent cells. In: Oncogene. 2014 ; Vol. 33, No. 4. pp. 440-448.
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abstract = "Intratumoral heterogeneity within individual breast tumors is a well-known phenomenon that may contribute to drug resistance. This heterogeneity is dependent on several factors, such as types of oncogenic drivers and tumor precursor cells. The purpose of our study was to engineer a mouse mammary tumor model with intratumoral heterogeneity by using defined genetic perturbations. To achieve this, we used mice with knockout (-/-) of Ink4a/Arf, a tumor suppressor locus; these mice are known to be susceptible to non-mammary tumors such as fibrosarcoma. To induce mammary tumors, we retrovirally introduced an oncogene, HRAS(G12V), into Ink4a/Arf-/- mammary cells in vitro, and those cells were inoculated into syngeneic mice mammary fat pads. We observed 100{\%} tumorigenesis. The tumors formed were negative for estrogen receptor, progesterone receptor and HER2. Further, they had pathological features similar to those of human triple-negative breast cancer (TNBC) (for example, pushing borders, central necrosis). The tumors were found to be heterogeneous and included two subpopulations: CD49f- quiescent cells and CD49f + cells. Contrary to our expectation, CD49f- quiescent cells had high tumor-initiating potential and CD49f+ cells had relatively low tumor-initiating potential. Gene expression analysis revealed that CD49f- quiescent cells overexpressed epithelial-to-mesenchymal transition-driving genes, reminiscent of tumor-initiating cells and claudin-low breast cancer. Our animal model with intratumoral heterogeneity, derived from defined genetic perturbations, allows us to test novel molecular targeted drugs in a setting that mimics the intratumoral heterogeneity of human TNBC.",
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