CoChat
Sign Up
Cited by 18Estrogen and related hormone effectsDevelopmental Biology and Gene RegulationBreast Cancer Treatment Studies

Transcriptional Repressor Tbx3 Is Required for the Hormone-Sensing Cell Lineage in Mammary Epithelium

Open access
Kunasegaran, Kamini, Ho, Victor, Chang, Ted H-. T., De Silva, Duvini, Bakker, Martijn L., Christoffels, Vincent M., Pietersen, Alexandra M.·2014·PLoS ONE
DOI 10.1371/journal.pone.0110191PMC PMC4208772
Open this paper in CoChat — free accountOpen PDF

Abstract

The transcriptional repressor Tbx3 is involved in lineage specification in several tissues during embryonic development. Germ-line mutations in the Tbx3 gene give rise to Ulnar-Mammary Syndrome (comprising reduced breast development) and Tbx3 is required for mammary epithelial cell identity in the embryo. Notably Tbx3 has been implicated in breast cancer, which develops in adult mammary epithelium, but the role of Tbx3 in distinct cell types of the adult mammary gland has not yet been characterized. Using a fluorescent reporter knock-in mouse, we show that in adult virgin mice Tbx3 is highly expressed in luminal cells that express hormone receptors, and not in luminal cells of the alveolar lineage (cells primed for milk production). Flow cytometry identified Tbx3 expression already in progenitor cells of the hormone-sensing lineage and co-immunofluorescence confirmed a strict correlation between estrogen receptor (ER) and Tbx3 expression in situ. Using in vivo reconstitution assays we demonstrate that Tbx3 is functionally relevant for this lineage because knockdown of Tbx3 in primary mammary epithelial cells prevented the formation of ER+ cells, but not luminal ER- or basal cells. Interestingly, genes that are repressed by Tbx3 in other cell types, such as E-cadherin, are not repressed in hormone-sensing cells, highlighting that transcriptional targets of Tbx3 are cell type specific. In summary, we provide the first analysis of Tbx3 expression in the adult mammary gland at a single cell level and show that Tbx3 is important for the generation of hormone-sensing cells.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0110191

Cites

5 of 5 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 44

  1. [1]Douglas NC, Papaioannou VE (2013) The T-box transcription factors TBX2 and TBX3 in mammary gland development and breast cancer. J Mammary Gland Biol Neoplasia 18: 143– 147 10.1007/s10911-013-9282-8 23624936 PMC3692603 · DOI
  2. [2]Propper AY, Howard BA, Veltmaat JM (2013) Prenatal morphogenesis of mammary glands in mouse and rabbit. J Mammary Gland Biol Neoplasia 18: 93– 104 10.1007/s10911-013-9298-0 23736987 PMC3691486 · DOI
  3. [3]Brisken C, O'Malley B (2010) Hormone action in the mammary gland. Cold Spring Harbor Perspectives in Biology 2: a003178 10.1101/cshperspect.a003178 20739412 PMC2982168 · DOI
  4. [4]Meier-Abt F, Bentires-Alj M (2014) How pregnancy at early age protects against breast cancer. Trends Mol Med 20: 143– 153 10.1016/j.molmed.2013.11.002 24355762 · DOI
  5. [5]Rios AC, Fu NY, Lindeman GJ, Visvader JE (2014) In situ identification of bipotent stem cells in the mammary gland. Nature 506: 322– 327 10.1038/nature12948 24463516 · DOI
  6. [7]van Amerongen R, Bowman AN, Nusse R (2012) Developmental stage and time dictate the fate of Wnt/β-catenin-responsive stem cells in the mammary gland. Cell Stem Cell 11: 387– 400 10.1016/j.stem.2012.05.023 22863533 PMC13155203 · DOI
  7. [8]Boulanger CA, Wagner K-U, Smith GH (2005) Parity-induced mouse mammary epithelial cells are pluripotent, self-renewing and sensitive to TGF-beta1 expression. Oncogene 24: 552– 560 10.1038/sj.onc.1208185 15580303 · DOI
  8. [9]Šale S, Lafkas D, Artavanis-Tsakonas S (2013) Notch2 genetic fate mapping reveals two previously unrecognized mammary epithelial lineages. Nat Cell Biol 15: 1– 11 10.1038/ncb2725 23604318 PMC4369920 · DOI
  9. [10]Chang TH-T, Kunasegaran K, Tarulli GA, De Silva D, Voorhoeve PM, et al (2014) New insights into lineage restriction of mammary gland epithelium using parity-identified mammary epithelial cells. Breast Cancer Res 16: R1 10.1186/bcr3593 24398145 PMC3978646 · DOI
  10. [11]Beleut M, Rajaram RD, Caikovski M, Ayyanan A, Germano D, et al (2010) Two distinct mechanisms underlie progesterone-induced proliferation in the mammary gland. Proceedings of the National Academy of Sciences 107: 2989– 2994 10.1073/pnas.0915148107 PMC2840294 20133621 · DOI
  11. [12]Mastroianni M, Kim S, Kim YC, Esch A, Wagner C, et al (2010) Wnt signaling can substitute for estrogen to induce division of ERα-positive cells in a mouse mammary tumor model. Cancer Lett 289: 23– 31 10.1016/j.canlet.2009.07.012 19665836 PMC2874254 · DOI
  12. [14]Regan JL, Kendrick H, Magnay F-A, Vafaizadeh V, Groner B, et al (2012) c-Kit is required for growth and survival of the cells of origin of Brca1-mutation-associated breast cancer. Oncogene 31: 869– 883 10.1038/onc.2011.289 21765473 · DOI
  13. [15]Hammond MEH, Hayes DF, Dowsett M, Allred DC, Hagerty KL, et al (2010) American Society of Clinical Oncology/College Of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. Journal of Clinical Oncology 28: 2784– 2795 10.1200/JCO.2009.25.6529 20404251 PMC2881855 · DOI
  14. [16]Santagata S, Thakkar A, Ergonul A, Wang B, Woo T, et al (2014) Taxonomy of breast cancer based on normal cell phenotype predicts outcome. J Clin Invest 124: 859– 870 10.1172/JCI70941 24463450 PMC3904619 · DOI
  15. [17]Davenport TG, Jerome-Majewska LA, Papaioannou VE (2003) Mammary gland, limb and yolk sac defects in mice lacking Tbx3, the gene mutated in human ulnar mammary syndrome. Development 130: 2263– 2273. 12668638
  16. [18]Jerome-Majewska LA, Jenkins GP, Ernstoff E, Zindy F, Sherr CJ, et al (2005) Tbx3, the ulnar-mammary syndrome gene, and Tbx2 interact in mammary gland development through a p19Arf/p53-independent pathway. Dev Dyn 234: 922– 933 10.1002/dvdy.20575 16222716 · DOI
  17. [19]Bamshad M, Lin RC, Law DJ, Watkins WC, Krakowiak PA, et al (1997) Mutations in human TBX3 alter limb, apocrine and genital development in ulnar-mammary syndrome. Nat Genet 16: 311– 315 10.1038/ng0797-311 9207801 · DOI · Paper page
  18. [20]Bamshad M, Le T, Watkins WS, Dixon ME, Kramer BE, et al (1999) The spectrum of mutations in TBX3: Genotype/Phenotype relationship in ulnar-mammary syndrome. Am J Hum Genet 64: 1550– 1562 10.1086/302417 10330342 PMC1377898 · DOI · Paper page
  19. [21]Washkowitz AJ, Gavrilov S, Begum S, Papaioannou VE (2012) Diverse functional networks of Tbx3 in development and disease. Wiley Interdiscip Rev Syst Biol Med 4: 273– 283 10.1002/wsbm.1162 22334480 PMC3328642 · DOI · Paper page
  20. [23]Brummelkamp TRT, Kortlever RMR, Lingbeek MM, Trettel FF, MacDonald MEM, et al (2002) TBX-3, the gene mutated in Ulnar-Mammary Syndrome, is a negative regulator of p19ARF and inhibits senescence. J Biol Chem 277: 6567– 6572 10.1074/jbc.M110492200 11748239 · DOI · Paper page
  21. [24]Rodriguez M, Aladowicz E, Lanfrancone L, Goding CR (2008) Tbx3 represses e-cadherin expression and enhances melanoma invasiveness. Cancer Res 68: 7872– 7881 10.1158/0008-5472.CAN-08-0301 18829543 · DOI · Paper page
  22. [26]Yarosh W, Barrientos T, Esmailpour T, Lin L, Carpenter PM, et al (2008) TBX3 is overexpressed in breast cancer and represses p14 ARF by interacting with histone deacetylases. Cancer Res 68: 693– 699 10.1158/0008-5472.CAN-07-5012 18245468 · DOI · Paper page
  23. [27]Fillmore CM, Gupta PB, Rudnick JA, Caballero S, Keller PJ, et al (2010) Estrogen expands breast cancer stem-like cells through paracrine FGF/Tbx3 signaling. Proc Natl Acad Sci USA 107: 21737– 21742 10.1073/pnas.1007863107 21098263 PMC3003123 · DOI
  24. [28]Tarulli GA, De Silva D, Ho V, Kunasegaran K, Ghosh K, et al (2013) Hormone-sensing cells require Wip1 for paracrine stimulation in normal and premalignant mammary epithelium. Breast Cancer Research 15: R10 10.1186/bcr3381 23369183 PMC3672744 · DOI
  25. [29]Ho V, Yeo SY, Kunasegaran K, De Silva D, Tarulli GA, et al (2013) Expression analysis of rare cellular subsets: direct RT-PCR on limited cell numbers obtained by FACS or soft agar assays. BioTechniques 54: 208– 212 10.2144/000114019 23581467 · DOI
  26. [30]Smalley MJ (2010) Isolation, culture and analysis of mouse mammary epithelial cells. Methods Mol Biol 633: 139– 170 10.1007/978-1-59745-019-511 20204626 · DOI
  27. [31]Oakes SR, Naylor MJ, Asselin-Labat M-L, Blazek KD, Gardiner-Garden M, et al (2008) The Ets transcription factor Elf5 specifies mammary alveolar cell fate. Genes Dev 22: 581– 586 10.1101/gad.1614608 18316476 PMC2259028 · DOI
  28. [32]Sleeman KE, Kendrick H, Robertson D, Isacke CM, Ashworth A, et al (2007) Dissociation of estrogen receptor expression and in vivo stem cell activity in the mammary gland. J Cell Biol 176: 19– 26 10.1083/jcb.200604065 17190790 PMC2063618 · DOI
  29. [33]Michalak EM, Nacerddine K, Pietersen A, Beuger V, Pawlitzky I, et al (2013) Polycomb group gene Ezh2 regulates mammary gland morphogenesis and maintains the luminal progenitor pool. STEM CELLS 31: 1910– 1920 10.1002/stem.1437 23712803 · DOI
  30. [34]Li M, Fu X, Ma G, Sun X, Dong X, et al (2012) Atbf1 Regulates Pubertal Mammary Gland Development Likely by Inhibiting the Pro-Proliferative Function of Estrogen-ER Signaling. PLoS ONE 7: e51283 10.1371/journal.pone.0051283.t001 23251482 PMC3520988 · DOI
  31. [35]Hoogaars WMH, Tessari A, Moorman AFM, de Boer PAJ, Hagoort J, et al (2004) The transcriptional repressor Tbx3 delineates the developing central conduction system of the heart. Cardiovasc Res 62: 489– 499 10.1016/j.cardiores.2004.01.030 15158141 · DOI
  32. [36]Hoogaars WMH, Engel A, Brons JF, Verkerk AO, de Lange FJ, et al (2007) Tbx3 controls the sinoatrial node gene program and imposes pacemaker function on the atria. Genes Dev 21: 1098– 1112 10.1101/gad.416007 17473172 PMC1855235 · DOI · Paper page
  33. [38]Eblaghie MC, Song S-J, Kim J-Y, Akita K, Tickle C, et al (2004) Interactions between FGF and Wnt signals and Tbx3 gene expression in mammary gland initiation in mouse embryos. J Anat 205: 1– 13 10.1111/j.0021-8782.2004.00309.x 15255957 PMC1571327 · DOI · Paper page
  34. [39]Parsa S, Ramasamy SK, De Langhe S, Gupte VV, Haigh JJ, et al (2008) Terminal end bud maintenance in mammary gland is dependent upon FGFR2b signaling. Dev Biol 317: 121– 131 10.1016/j.ydbio.2008.02.014 18381212 · DOI
  35. [40]Lu P, Ewald AJ, Martin GR, Werb Z (2008) Genetic mosaic analysis reveals FGF receptor 2 function in terminal end buds during mammary gland branching morphogenesis. Dev Biol 321: 77– 87 10.1016/j.ydbio.2008.06.005 18585375 PMC2582391 · DOI
  36. [41]Ewan KBR, Oketch-Rabah HA, Ravani SA, Shyamala G, Moses HL, et al (2005) Proliferation of estrogen receptor-alpha-positive mammary epithelial cells is restrained by transforming growth factor-beta1 in adult mice. Am J Pathol 167: 409– 417. 16049327 PMC1603552
  37. [42]Lingbeek ME, Jacobs JJL, van Lohuizen M (2002) The T-box repressors TBX2 and TBX3 specifically regulate the tumor suppressor gene p14ARF via a variant T-site in the initiator. J Biol Chem 277: 26120– 26127 10.1074/jbc.M200403200 12000749 · DOI · Paper page
  38. [43]van den Boogaard M, Wong LYE, Tessadori F, Bakker ML, Dreizehnter LK, et al (2012) Genetic variation in T-box binding element functionally affects SCN5A/SCN10A enhancer. J Clin Invest 122: 2519– 2530 10.1172/JCI62613 22706305 PMC3386824 · DOI
  39. [44]Han J, Yuan P, Yang H, Zhang J, Soh BS, et al (2010) Tbx3 improves the germ-line competency of induced pluripotent stem cells. Nature 463: 1096– 1100 10.1038/nature08735 20139965 PMC2901797 · DOI · Paper page
  40. [45]Hoogaars WMH, Barnett P, Rodriguez M, Clout DE, Moorman AFM, et al (2008) TBX3 and its splice variant TBX3 + exon 2a are functionally similar. Pigment Cell Melanoma Res 21: 379– 387 10.1111/j.1755-148X.2008.00461.x 18444963 · DOI
  41. [46]Stephens PJ, Tarpey PS, Davies H, Van Loo P, Greenman C, et al (2012) The landscape of cancer genes and mutational processes in breast cancer. Nature 486: 400– 404 10.1038/nature11017 22722201 PMC3428862 · DOI
  42. [47]Ellis MJ, Ding L, Shen D, Luo J, Suman VJ, et al. (2012) Whole-genome analysis informs breast cancer response to aromatase inhibition. Nature: 1–8. doi:10.1038/nature11143. PMC3383766 22722193 · DOI
  43. [48]Peres J, Davis E, Mowla S, Bennett DC, Li JA, et al (2010) The Highly Homologous T-Box Transcription Factors, TBX2 and TBX3, Have Distinct Roles in the Oncogenic Process. Genes & Cancer 1: 272– 282 10.1177/1947601910365160 21779450 PMC3092191 · DOI
  44. [49]Lu J, Li X-P, Dong Q, Kung H-F, He M-L (2010) TBX2 and TBX3: the special value for anticancer drug targets. Biochim Biophys Acta 1806: 268– 274 10.1016/j.bbcan.2010.07.001 20624445 PMC7127380 · DOI · Paper page

Cited by

Sorted by citations

Authors
Kunasegaran, K., Ho, V., Chang, T. H. T., De Silva, D., Bakker, M. L., Christoffels, V. M., & Pietersen, A. M.
Affiliations
  • National University of Singapore (SG)
  • Amsterdam UMC Location University of Amsterdam (NL)
Corresponding author
Pietersen, Alexandra M.
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2014-10-24
Volume
9
Issue
10
Pages
e110191
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0110191
PMC
PMC4208772
OpenAlex
W1984530325
Cited by
18
References
5
Citations by year
2024: 5 · 2023: 2 · 2022: 1 · 2021: 2 · 2018: 6 · 2015: 2
FWCI
0.44
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • National University of Singapore (submittedVersion)
  • Data Archiving and Networked Services (DANS) (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
  • Pure Amsterdam UMC (submittedVersion)
Funders
  • Duke-NUS Medical School
  • Agency for Science, Technology and Research
  • Shanxi Scholarship Council of China
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Breast Cancer Treatment Studies
Keywords
TBX3, transcriptional repressor, mammary epithelium, luminal cells, estrogen receptor, alveolar cells, mammary progenitor cells, E-cadherin, ulnar-mammary syndrome, breast cancer, single-cell gene expression
MeSH terms
Aging, Animals, Animals, Newborn, Epithelial Cells, Female, Fluorescence, Hormones, Mammary Glands, Animal, Receptors, Estrogen, Repressor Proteins, Transcription, Genetic, Genes, Reporter, Cell Lineage, T-Box Domain Proteins, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review