Abstract
In the mammary gland, it is widely believed that the luminal cells are unipotent after birth, contributing only to the luminal compartment in normal development. Here, by lineage tracing, we uncovered an unexpected potential of luminal cells that can give rise to basal cells during pregnancy. These luminal-derived basal cells (LdBCs) persisted through mammary regression and generated more progeny in successive rounds of pregnancies. LdBCs express basal markers as well as estrogen receptor α (ERα). In ovariectomized (OVX) mice, stimulation with estrogen and progesterone promoted the formation of LdBCs. In serial transplantation assays, LdBCs were able to reconstitute new mammary glands in a hormone-dependent manner. Transcriptome analysis and genetic experiments suggest that Wnt/β-catenin signaling is essential for the formation and maintenance of LdBCs. Our data uncover an unexpected bi-potency of luminal cells in a physiological context. The discovery of ERα+ basal cells, which can respond to hormones and are endowed with stem cell-like regenerative capacity in parous mammary gland, provides new insights into the association of hormones and breast cancer.
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26 February 2019
In the initial published version of this article, there was a mistake in one author name (Jingsong Li). The correct name should be “Jinsong Li”. This correction does not affect the description of the results or the conclusions of this work.
References
Sreekumar, A., Roarty, K. & Rosen, J. M. The mammary stem cell hierarchy: a looking glass into heterogeneous breast cancer landscapes. Endocr. Relat. Cancer 22, T161–T176 (2015).
Visvader, J. E. & Stingl, J. Mammary stem cells and the differentiation hierarchy: current status and perspectives. Genes Dev. 28, 1143–1158 (2014).
Stingl, J. et al. Purification and unique properties of mammary epithelial stem cells. Nature 439, 993–997 (2006).
Shackleton, M. et al. Generation of a functional mammary gland from a single stem cell. Nature 439, 84–88 (2006).
Rios, A. C., Fu, N. Y., Lindeman, G. J. & Visvader, J. E. In situ identification of bipotent stem cells in the mammary gland. Nature 506, 322–327 (2014).
Wang, D. et al. Identification of multipotent mammary stem cells by protein C receptor expression. Nature 517, 81–84 (2015).
Van Keymeulen, A. et al. Distinct stem cells contribute to mammary gland development and maintenance. Nature 479, 189–193 (2011).
van Amerongen, R., Bowman, A. N. & Nusse, R. Developmental stage and time dictate the fate of Wnt/beta-catenin−responsive stem cells in the mammary gland. Cell Stem Cell 11, 387–400 (2012).
Prater, M. D. et al. Mammary stem cells have myoepithelial cell properties. Nat. Cell Biol. 16, 942–950 (2014). 941–947.
de Visser, K. E. et al. Developmental stage-specific contribution of LGR5(+) cells to basal and luminal epithelial lineages in the postnatal mammary gland. J. Pathol. 228, 300–309 (2012).
Wuidart, A. et al. Quantitative lineage tracing strategies to resolve multipotency in tissue-specific stem cells. Genes Dev. 30, 1261–1277 (2016).
Davis, F. M. et al. Single-cell lineage tracing in the mammary gland reveals stochastic clonal dispersion of stem/progenitor cell progeny. Nat. Commun. 7, 13053 (2016).
Brisken, C. & O’Malley, B. Hormone action in the mammary gland. Cold Spring Harb. Perspect. Biol. 2, a003178 (2010).
Clarke, R. B., Howell, A., Potten, C. S. & Anderson, E. Dissociation between steroid receptor expression and cell proliferation in the human breast. Cancer Res 57, 4987–4991 (1997).
Shyamala, G. et al. Cellular expression of estrogen and progesterone receptors in mammary glands: regulation by hormones, development and aging. J. Steroid Biochem Mol. Biol. 80, 137–148 (2002).
Petersen, O. W., Hoyer, P. E. & van Deurs, B. Frequency and distribution of estrogen receptor-positive cells in normal, nonlactating human breast tissue. Cancer Res. 47, 5748–5751 (1987).
Lim, E. et al. Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers. Nat. Med. 15, 907–913 (2009).
Asselin-Labat, M. L. et al. Steroid hormone receptor status of mouse mammary stem cells. J. Natl Cancer Inst. 98, 1011–1014 (2006).
Sleeman, K. E. et al. Dissociation of estrogen receptor expression and in vivo stem cell activity in the mammary gland. J. Cell Biol. 176, 19–26 (2007).
Stingl, J. Estrogen and progesterone in normal mammary gland development and in cancer. Horm. Cancer 2, 85–90 (2011).
Bergkvist, L., Adami, H. O., Persson, I., Hoover, R. & Schairer, C. The risk of breast cancer after estrogen and estrogen-progestin replacement. N. Engl. J. Med 321, 293–297 (1989).
Pike, M. C., Spicer, D. V., Dahmoush, L. & Press, M. F. Estrogens, progestogens, normal breast cell proliferation, and breast cancer risk. Epidemiol. Rev. 15, 17–35 (1993).
Clemons, M. & Goss, P. Estrogen and the risk of breast cancer. N. Engl. J. Med 344, 276–285 (2001).
Chang, T. H. et al. New insights into lineage restriction of mammary gland epithelium using parity-identified mammary epithelial cells. Breast Cancer Res. 16, R1 (2014).
Rodilla, V. et al. Luminal progenitors restrict their lineage potential during mammary gland development. PLoS Biol. 13, e1002069 (2015).
Van Keymeulen, A. et al. Lineage-restricted mammary stem cells sustain the development, homeostasis, and regeneration of the estrogen receptor positive lineage. Cell Rep. 20, 1525–1532 (2017).
Wang, C., Christin, J. R., Oktay, M. H. & Guo, W. Lineage-biased stem cells maintain estrogen-receptor-positive and -negative mouse mammary luminal lineages. Cell Rep. 18, 2825–2835 (2017).
Elias, S., Morgan, M. A., Bikoff, E. K. & Robertson, E. J. Long-lived unipotent Blimp1-positive luminal stem cells drive mammary gland organogenesis throughout adult life. Nat. Commun. 8, 1714 (2017).
Lafkas, D. et al. Notch3 marks clonogenic mammary luminal progenitor cells in vivo. J. Cell Biol. 203, 47–56 (2013).
Bach, K. et al. Differentiation dynamics of mammary epithelial cells revealed by single-cell RNA sequencing. Nat. Commun. 8, 2128 (2017).
Pal, B. et al. Construction of developmental lineage relationships in the mouse mammary gland by single-cell RNA profiling. Nat. Commun. 8, 1627 (2017).
Guo, W. et al. Slug and Sox9 cooperatively determine the mammary stem cell state. Cell 148, 1015–1028 (2012).
Panciera, T. et al. Induction of expandable tissue-specific stem/progenitor cells through transient expression of YAP/TAZ. Cell Stem Cell 19, 725–737 (2016).
Molyneux, G. et al. BRCA1 basal-like breast cancers originate from luminal epithelial progenitors and not from basal stem cells. Cell Stem Cell 7, 403–417 (2010).
Ling, H. & Jolicoeur, P. Notch-1 signaling promotes the cyclinD1-dependent generation of mammary tumor-initiating cells that can revert to bi-potential progenitors from which they arise. Oncogene 32, 3410–3419 (2013).
Hein, S. M. et al. Luminal epithelial cells within the mammary gland can produce basal cells upon oncogenic stress. Oncogene 35, 1461–1467 (2016).
Koren, S. et al. PIK3CA(H1047R) induces multipotency and multi-lineage mammary tumours. Nature 525, 114–118 (2015).
Van Keymeulen, A. et al. Reactivation of multipotency by oncogenic PIK3CA induces breast tumour heterogeneity. Nature 525, 119–123 (2015).
Zhang, L. et al. Targeting CreER(T2) expression to keratin 8-expressing murine simple epithelia using bacterial artificial chromosome transgenesis. Transgenic Res. 21, 1117–1123 (2012).
Choi, N., Zhang, B., Zhang, L., Ittmann, M. & Xin, L. Adult murine prostate basal and luminal cells are self-sustained lineages that can both serve as targets for prostate cancer initiation. Cancer Cell 21, 253–265 (2012).
Yu, Q. C., Verheyen, E. M. & Zeng, Y. A. Mammary development and breast cancer: a Wnt perspective. Cancers (Basel) 8 https://doi.org/10.3390/cancers8070065 (2016).
Markopoulos, C., Berger, U., Wilson, P., Gazet, J. C. & Coombes, R. C. Oestrogen receptor content of normal breast cells and breast carcinomas throughout the menstrual cycle. Br. Med J. 296, 1349–1351 (1988).
Press, M. F., Nousek-Goebl, N., King, W. J., Herbst, A. L. & Greene, G. L. Immunohistochemical assessment of estrogen receptor distribution in the human endometrium throughout the menstrual cycle. Lab Invest. 51, 495–503 (1984).
Asselin-Labat, M. L. et al. Control of mammary stem cell function by steroid hormone signalling. Nature 465, 798–802 (2010).
Ellison-Zelski, S. J., Solodin, N. M. & Alarid, E. T. Repression of ESR1 through actions of estrogen receptor alpha and Sin3A at the proximal promoter. Mol. Cell Biol. 29, 4949–4958 (2009).
Niemann, C., Owens, D. M., Hulsken, J., Birchmeier, W. & Watt, F. M. Expression of DeltaNLef1 in mouse epidermis results in differentiation of hair follicles into squamous epidermal cysts and formation of skin tumours. Development 129, 95–109 (2002).
Harada, N. et al. Intestinal polyposis in mice with a dominant stable mutation of the beta-catenin gene. EMBO J. 18, 5931–5942 (1999).
Lindley, L. E. et al. The WNT-controlled transcriptional regulator LBH is required for mammary stem cell expansion and maintenance of the basal lineage. Development 142, 893–904 (2015).
Chakrabarti, R. et al. DeltaNp63 promotes stem cell activity in mammary gland development and basal-like breast cancer by enhancing Fzd7 expression and Wnt signalling. Nat. Cell Biol. 16, 1004–1015, 1001–1013 (2014).
Gu, B., Watanabe, K., Sun, P., Fallahi, M. & Dai, X. Chromatin effector Pygo2 mediates Wnt-notch crosstalk to suppress luminal/alveolar potential of mammary stem and basal cells. Cell Stem Cell 13, 48–61 (2013).
Bouras, T. et al. Notch signaling regulates mammary stem cell function and luminal cell-fate commitment. Cell Stem Cell 3, 429–441 (2008).
Wagner, K. U. et al. An adjunct mammary epithelial cell population in parous females: its role in functional adaptation and tissue renewal. Development 129, 1377–1386 (2002).
Matulka, L. A., Triplett, A. A. & Wagner, K. U. Parity-induced mammary epithelial cells are multipotent and express cell surface markers associated with stem cells. Dev. Biol. 303, 29–44 (2007).
Boulanger, C. A., Wagner, K. U. & Smith, G. H. Parity-induced mouse mammary epithelial cells are pluripotent, self-renewing and sensitive to TGF-beta1 expression. Oncogene 24, 552–560 (2005).
Keller, P. J. et al. Defining the cellular precursors to human breast cancer. Proc. Natl Acad. Sci. USA 109, 2772–2777 (2012).
Ince, T. A. et al. Transformation of different human breast epithelial cell types leads to distinct tumor phenotypes. Cancer Cell 12, 160–170 (2007).
Gastaldi, S. et al. Met signaling regulates growth, repopulating potential and basal cell-fate commitment of mammary luminal progenitors: implications for basal-like breast cancer. Oncogene 32, 1428–1440 (2013).
Janerich, D. T. & Hoff, M. B. Evidence for a crossover in breast cancer risk factors. Am. J. Epidemiol. 116, 737–742 (1982).
Schedin, P. Pregnancy-associated breast cancer and metastasis. Nat. Rev. Cancer 6, 281–291 (2006).
MacMahon, B. et al. Age at first birth and breast cancer risk. Bull. World Health Organ 43, 209–221 (1970).
Muzumdar, M. D., Tasic, B., Miyamichi, K., Li, L. & Luo, L. A global double-fluorescent Cre reporter mouse. Genesis 45, 593–605 (2007).
Rinkevich, Y., Lindau, P., Ueno, H., Longaker, M. T. & Weissman, I. L. Germ-layer and lineage-restricted stem/progenitors regenerate the mouse digit tip. Nature 476, 409–413 (2011).
Huelsken, J., Vogel, R., Erdmann, B., Cotsarelis, G. & Birchmeier, W. beta-Catenin controls hair follicle morphogenesis and stem cell differentiation in the skin. Cell 105, 533–545 (2001).
Chen, J. et al. Spatial transcriptomic analysis of cryosectioned tissue samples with Geo-seq. Nat. Protoc. 12, 566–580 (2017).
Trapnell, C., Pachter, L. & Salzberg, S. L. TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25, 1105–1111 (2009).
Kim, D. et al. TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol. 14, R36 (2013).
Li, Y. Y. et al. Transcriptome analysis reveals determinant stages controlling human embryonic stem cell commitment to neuronal cells. J. Biol. Chem. 292, 19590–19604 (2017).
Sozen, B. et al. Self-assembly of embryonic and two extra- embryonic stem cell types into gastrulating embryo-like structures. Nat. Cell Biol. 20, 979 (2018).
Hong, F. X. et al. RankProd: a bioconductor package for detecting differentially expressed genes in meta-analysis. Bioinformatics 22, 2825–2827 (2006).
Saldanha, A. J. Java Treeview-extensible visualization of microarray data. Bioinformatics 20, 3246–3248 (2004).
Yang, X. et al. TGFbeta signaling hyperactivation-induced tumorigenicity during the derivation of neural progenitors from mouse ESCs. J. Mol. Cell Biol. 10, 216–228 (2018).
Huang da, W., Sherman, B. T. & Lempicki, R. A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat. Protoc. 4, 44–57 (2009).
Acknowledgements
We thank Dr. Li Xin for kindly sharing the K8-CreERT2 mouse line and Dr. Fiona Watt for kindly sharing the K14-Lef1Δn mouse line. This work was supported by grants from National Natural Science Foundation of China (31830056, 31861163006, 31625020, 31530045, 31661143043 to Y.A.Z.; 31871456, 31661143042, 91519314, 31630043, 31571513, 31430058 to N.J.), Chinese Academy of Sciences (XDB19000000 and XDA12020349 to Y.A.Z.; XDA16020501 to N.J., XDA16020404 to G.P.), Ministry of Science and Technology of China (2018YFA0108000, 2017YFA0102700, 2015CB964500 to N.J., 2018YFA0107201 to G.P.), and Research Grants Council (N_HKUST628/18 to T.H.C), Shanghai Municipal Science and Technology Commission (17XD1404000 to Y.A.Z.).
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W.S. and Y.A.Z. designed the experiments. W.S. and R.Y. performed immunostaining staining, FACS and transplantation experiments. R.W. and G.P. performed RNA sequencing and data analysis, T.C. and N.J. supervised bioinformatics analysis. W.S., W.J. and Q.C.Y. generated mouse strains and conducted genetic experiments. Q.Y. and J.L. redirived and generated the rainbow-reporter mouse strain. W.S., R.W. and Y.A.Z. analyzed the data and wrote the manuscript.
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Song, W., Wang, R., Jiang, W. et al. Hormones induce the formation of luminal-derived basal cells in the mammary gland. Cell Res 29, 206–220 (2019). https://doi.org/10.1038/s41422-018-0137-0
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DOI: https://doi.org/10.1038/s41422-018-0137-0
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