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Breast Cancer Research
Published in: Breast Cancer Research 1/2018

Open Access 01-12-2018 | Research article

NOTCH3 expression is linked to breast cancer seeding and distant metastasis

Authors: Alexey A. Leontovich, Mohammad Jalalirad, Jeffrey L. Salisbury, Lisa Mills, Candace Haddox, Mark Schroeder, Ann Tuma, Maria E. Guicciardi, Luca Zammataro, Mario W. Gambino, Angela Amato, Aldo Di Leonardo, James McCubrey, Carol A. Lange, Minetta Liu, Tufia Haddad, Matthew Goetz, Judy Boughey, Jann Sarkaria, Liewei Wang, James N. Ingle, Evanthia Galanis, Antonino B. D’Assoro

Published in: Breast Cancer Research | Issue 1/2018

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Abstract

Background

Development of distant metastases involves a complex multistep biological process termed the invasion-metastasis cascade, which includes dissemination of cancer cells from the primary tumor to secondary organs. NOTCH developmental signaling plays a critical role in promoting epithelial-to-mesenchymal transition, tumor stemness, and metastasis. Although all four NOTCH receptors show oncogenic properties, the unique role of each of these receptors in the sequential stepwise events that typify the invasion-metastasis cascade remains elusive.

Methods

We have established metastatic xenografts expressing high endogenous levels of NOTCH3 using estrogen receptor alpha-positive (ERα+) MCF-7 breast cancer cells with constitutive active Raf-1/mitogen-associated protein kinase (MAPK) signaling (vMCF-7Raf-1) and MDA-MB-231 triple-negative breast cancer (TNBC) cells. The critical role of NOTCH3 in inducing an invasive phenotype and poor outcome was corroborated in unique TNBC cells resulting from a patient-derived brain metastasis (TNBC-M25) and in publicly available claudin-low breast tumor specimens collected from participants in the Molecular Taxonomy of Breast Cancer International Consortium database.

Results

In this study, we identified an association between NOTCH3 expression and development of metastases in ERα+ and TNBC models. ERα+ breast tumor xenografts with a constitutive active Raf-1/MAPK signaling developed spontaneous lung metastases through the clonal expansion of cancer cells expressing a NOTCH3 reprogramming network. Abrogation of NOTCH3 expression significantly reduced the self-renewal and invasive capacity of ex vivo breast cancer cells, restoring a luminal CD44low/CD24high/ERαhigh phenotype. Forced expression of the mitotic Aurora kinase A (AURKA), which promotes breast cancer metastases, failed to restore the invasive capacity of NOTCH3-null cells, demonstrating that NOTCH3 expression is required for an invasive phenotype. Likewise, pharmacologic inhibition of NOTCH signaling also impaired TNBC cell seeding and metastatic growth. Significantly, the role of aberrant NOTCH3 expression in promoting tumor self-renewal, invasiveness, and poor outcome was corroborated in unique TNBC cells from a patient-derived brain metastasis and in publicly available claudin-low breast tumor specimens.

Conclusions

These findings demonstrate the key role of NOTCH3 oncogenic signaling in the genesis of breast cancer metastasis and provide a compelling preclinical rationale for the design of novel therapeutic strategies that will selectively target NOTCH3 to halt metastatic seeding and to improve the clinical outcomes of patients with breast cancer.
Appendix
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Metadata
Title
NOTCH3 expression is linked to breast cancer seeding and distant metastasis
Authors
Alexey A. Leontovich
Mohammad Jalalirad
Jeffrey L. Salisbury
Lisa Mills
Candace Haddox
Mark Schroeder
Ann Tuma
Maria E. Guicciardi
Luca Zammataro
Mario W. Gambino
Angela Amato
Aldo Di Leonardo
James McCubrey
Carol A. Lange
Minetta Liu
Tufia Haddad
Matthew Goetz
Judy Boughey
Jann Sarkaria
Liewei Wang
James N. Ingle
Evanthia Galanis
Antonino B. D’Assoro
Publication date
01-12-2018
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 1/2018
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/s13058-018-1020-0

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