Abstract
Basal-like breast cancers arising in women carrying mutations in the BRCA1 gene, encoding the tumor suppressor protein BRCA1, are thought to develop from the mammary stem cell. To explore early cellular changes that occur in BRCA1 mutation carriers, we have prospectively isolated distinct epithelial subpopulations from normal mammary tissue and preneoplastic specimens from individuals heterozygous for a BRCA1 mutation. We describe three epithelial subsets including basal stem/progenitor, luminal progenitor and mature luminal cells. Unexpectedly, we found that breast tissue from BRCA1 mutation carriers harbors an expanded luminal progenitor population that shows factor-independent growth in vitro. Moreover, gene expression profiling revealed that breast tissue heterozygous for a BRCA1 mutation and basal breast tumors were more similar to normal luminal progenitor cells than any other subset, including the stem cell–enriched population. The c-KIT tyrosine kinase receptor (encoded by KIT) emerged as a key marker of luminal progenitor cells and was more highly expressed in BRCA1-associated preneoplastic tissue and tumors. Our findings suggest that an aberrant luminal progenitor population is a target for transformation in BRCA1-associated basal tumors .
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Turner, N., Tutt, A. & Ashworth, A. Hallmarks of 'BRCAness' in sporadic cancers. Nat. Rev. Cancer 4, 814–819 (2004).
Herschkowitz, J.I. et al. Identification of conserved gene expression features between murine mammary carcinoma models and human breast tumors. Genome Biol. 8, R76 (2007).
Perou, C.M. et al. Molecular portraits of human breast tumours. Nature 406, 747–752 (2000).
Foulkes, W.D. BRCA1 functions as a breast stem cell regulator. J. Med. Genet. 41, 1–5 (2004).
Ganesan, S. et al. Abnormalities of the inactive X chromosome are a common feature of BRCA1 mutant and sporadic basal-like breast cancer. Cold Spring Harb. Symp. Quant. Biol. 70, 93–97 (2005).
Narod, S.A. & Foulkes, W.D. BRCA1 and BRCA2: 1994 and beyond. Nat. Rev. Cancer 4, 665–676 (2004).
Venkitaraman, A.R. Cancer susceptibility and the functions of BRCA1 and BRCA2. Cell 108, 171–182 (2002).
Xu, X. et al. Conditional mutation of Brca1 in mammary epithelial cells results in blunted ductal morphogenesis and tumour formation. Nat. Genet. 22, 37–43 (1999).
Bouwman, P. & Jonkers, J. Mouse models for BRCA1 associated tumorigenesis: from fundamental insights to preclinical utility. Cell Cycle 7, 2647–2653 (2008).
Furuta, S. et al. Depletion of BRCA1 impairs differentiation but enhances proliferation of mammary epithelial cells. Proc. Natl. Acad. Sci. USA 102, 9176–9181 (2005).
Kubista, M., Rosner, M., Kubista, E., Bernaschek, G. & Hengstschlager, M. Brca1 regulates in vitro differentiation of mammary epithelial cells. Oncogene 21, 4747–4756 (2002).
Liu, S. et al. BRCA1 regulates human mammary stem/progenitor cell fate. Proc. Natl. Acad. Sci. USA 105, 1680–1685 (2008).
Clarke, R.B. et al. A putative human breast stem cell population is enriched for steroid receptor-positive cells. Dev. Biol. 277, 443–456 (2005).
Dontu, G. et al. In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev. 17, 1253–1270 (2003).
Gudjonsson, T. et al. Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. Genes Dev. 16, 693–706 (2002).
Raouf, A. et al. Transcriptome analysis of the normal human mammary cell commitment and differentiation process. Cell Stem Cell 3, 109–118 (2008).
Stingl, J., Eaves, C.J., Zandieh, I. & Emerman, J.T. Characterization of bipotent mammary epithelial progenitor cells in normal adult human breast tissue. Breast Cancer Res. Treat. 67, 93–109 (2001).
Villadsen, R. et al. Evidence for a stem cell hierarchy in the adult human breast. J. Cell Biol. 177, 87–101 (2007).
Ginestier, C. et al. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell 1, 555–567 (2007).
Eirew, P. et al. A method for quantifying normal human mammary epithelial stem cells with in vivo regenerative ability. Nat. Med. 14, 1384–1389 (2008).
Shackleton, M. et al. Generation of a functional mammary gland from a single stem cell. Nature 439, 84–88 (2006).
Stingl, J. et al. Purification and unique properties of mammary epithelial stem cells. Nature 439, 993–997 (2006).
Gusterson, B.A., Ross, D.T., Heath, V.J. & Stein, T. Basal cytokeratins and their relationship to the cellular origin and functional classification of breast cancer. Breast Cancer Res. 7, 143–148 (2005).
Nagle, R.B. et al. Characterization of breast carcinomas by two monoclonal antibodies distinguishing myoepithelial from luminal epithelial cells. J. Histochem. Cytochem. 34, 869–881 (1986).
Kuperwasser, C. et al. Reconstruction of functionally normal and malignant human breast tissues in mice. Proc. Natl. Acad. Sci. USA 101, 4966–4971 (2004).
Asselin-Labat, M.L. et al. Gata-3 is an essential regulator of mammary-gland morphogenesis and luminal-cell differentiation. Nat. Cell Biol. 9, 201–209 (2007).
Romijn, H.J., van Huizen, F. & Wolters, P.S. Towards an improved serum-free, chemically defined medium for long-term culturing of cerebral cortex tissue. Neurosci. Biobehav. Rev. 8, 301–334 (1984).
Ma, Y. et al. The breast cancer susceptibility gene BRCA1 regulates progesterone receptor signaling in mammary epithelial cells. Mol. Endocrinol. 20, 14–34 (2006).
Poole, A.J. et al. Prevention of Brca1-mediated mammary tumorigenesis in mice by a progesterone antagonist. Science 314, 1467–1470 (2006).
Michaud, J. et al. Integrative analysis of RUNX1 downstream pathways and target genes. BMC Genomics 9, 363 (2008).
Nielsen, T.O. et al. Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma. Clin. Cancer Res. 10, 5367–5374 (2004).
Simon, R. et al. KIT (CD117)-positive breast cancers are infrequent and lack KIT gene mutations. Clin. Cancer Res. 10, 178–183 (2004).
Asselin-Labat, M.L. et al. Steroid hormone receptor status of mouse mammary stem cells. J. Natl. Cancer Inst. 98, 1011–1014 (2006).
Kauff, N.D. et al. Risk-reducing salpingo-oophorectomy for the prevention of BRCA1- and BRCA2-associated breast and gynecologic cancer: a multicenter, prospective study. J. Clin. Oncol. 26, 1331–1337 (2008).
Kauff, N.D. et al. Risk-reducing salpingo-oophorectomy in women with a BRCA1 or BRCA2 mutation. N. Engl. J. Med. 346, 1609–1615 (2002).
Rebbeck, T.R. et al. Prophylactic oophorectomy in carriers of BRCA1 or BRCA2 mutations. N. Engl. J. Med. 346, 1616–1622 (2002).
Narod, S.A. Modifiers of risk of hereditary breast cancer. Oncogene 25, 5832–5836 (2006).
Mann, G.J. et al. Analysis of cancer risk and BRCA1 and BRCA2 mutation prevalence in the kConFab familial breast cancer resource. Breast Cancer Res. 8, R12 (2006).
Shultz, L.D. et al. Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2Rγ null mice engrafted with mobilized human hemopoietic stem cells. J. Immunol. 174, 6477–6489 (2005).
Wagner, K.U. et al. Cre-mediated gene deletion in the mammary gland. Nucleic Acids Res. 25, 4323–4330 (1997).
Deome, K.B., Faulkin, L.J. Jr., Bern, H.A. & Blair, P.B. Development of mammary tumors from hyperplastic alveolar nodules transplanted into gland-free mammary fat pads of female C3H mice. Cancer Res. 19, 515–520 (1959).
Laidlaw, I.J. et al. The proliferation of normal human breast tissue implanted into athymic nude mice is stimulated by estrogen but not progesterone. Endocrinology 136, 164–171 (1995).
Gentleman, R.C. et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80 (2004).
Smyth, G.K. Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Mol. Biol. 3, Article3 (2004).
Acknowledgements
We thank K. Stoev and K. Johnson for excellent animal husbandry, S. Mihajlovic and E. Tsui for expert assistance with histology and F. Battye and his colleagues for expert help in the flow cytometry lab. We thank J. Sambrook, E. McGowan, E. Musgrove and J. Adams for invaluable discussions and R. Reddel (Children's Medical Research Institute) for hTERT-immortalized fibroblasts. We thank K.U. Wagner (University of Nebraska Medical Center) for MMTV-Cre mice and A. Parlow (National Hormone and Pituitary Program, US National Institute of Diabetes, Digestive and Kidney Diseases) for prolactin. We gratefully acknowledge the invaluable contribution of numerous patients, surgeons, pathologists and tissue bank coordinators, and we thank A. Willems, E. Niedermayr, all kConFab research staff and Family Cancer Clinics and Clinical Follow-Up Study for their contributions to the kConFab resource, as well as the many families who contribute to kConFab. This work was supported by the Victorian Breast Cancer Research Consortium, the Australian National Health and Medical Research Council, the US National Breast Cancer Foundation, the US Department of Defense, the Susan G. Komen Breast Cancer Foundation, the Australian Stem Cell Centre, the Australian Cancer Research Foundation and the Victorian Cancer Biobank. kConFab is supported by grants from the National Breast Cancer Foundation, the National Health and Medical Research Council, the Queensland Cancer Fund, the Cancer Councils of New South Wales, Victoria, Tasmania and South Australia, and the Cancer Foundation of Western Australia.
Author information
Authors and Affiliations
Consortia
Contributions
E.L., F.V. and N.C.F. conducted most of the experiments and contributed to the writing of the manuscript. D.W. and G.K.S. performed the bioinformatic analyses and contributed to the writing of the manuscript. B.P, A.H.H. and M.-L.A.-L. performed RNA studies. D.E.G. and T.W. contributed to tissue preparation, immunohistochemistry and cell culture. F.F. helped optimize and performed some of the immunohistochemistry. A.P., H.J.T. and kConFab helped organize the accrual of the human breast tissue material. L.I.H. generated the hTERT-immortalized fibroblasts used for xenotransplantation studies. S.B.F. and M.Y. contributed to c-KIT staining and scoring. J.D.F. and M.A.B. contributed to the Brca1 experiments in mice. J.E.V. and G.J.L. conceptualized the study, contributed to study design and drafted and finalized the writing of the manuscript.
Corresponding authors
Additional information
The Kathleen Cuningham Consortium for Research into Familial Breast Cancer.
Supplementary information
Supplementary Text and Figures
Supplementary Methods, Supplementary Tables 1–4 and Supplementary Figs. 1–9 (PDF 2755 kb)
Supplementary Table 5
MaSC-enriched signature (XLS 300 kb)
Supplementary Table 6
Luminal progenitor gene signature (XLS 90 kb)
Supplementary Table 7
Luminal mature gene signature (XLS 138 kb)
Supplementary Table 8
Stroma signature (XLS 214 kb)
Rights and permissions
About this article
Cite this article
Lim, E., Vaillant, F., Wu, D. et al. Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers. Nat Med 15, 907–913 (2009). https://doi.org/10.1038/nm.2000
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm.2000
This article is cited by
-
Fatty acid amide hydrolase drives adult mammary gland development by promoting luminal cell differentiation
Cell Death Discovery (2024)
-
Identification of aberrant luminal progenitors and mTORC1 as a potential breast cancer prevention target in BRCA2 mutation carriers
Nature Cell Biology (2024)
-
An exploratory study for tuft cells in the breast and their relevance in triple-negative breast cancer: the possible relationship of SOX9
BMC Cancer (2023)
-
Claudin-4-adhesion signaling drives breast cancer metabolism and progression via liver X receptor β
Breast Cancer Research (2023)
-
COMMD3 loss drives invasive breast cancer growth by modulating copper homeostasis
Journal of Experimental & Clinical Cancer Research (2023)