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A stemness-related ZEB1–MSRB3 axis governs cellular pliancy and breast cancer genome stability


Chromosomal instability (CIN), a feature of most adult neoplasms from their early stages onward, is a driver of tumorigenesis. However, several malignancy subtypes, including some triple-negative breast cancers, display a paucity of genomic aberrations, thus suggesting that tumor development may occur in the absence of CIN. Here we show that the differentiation status of normal human mammary epithelial cells dictates cell behavior after an oncogenic event and predetermines the genetic routes toward malignancy. Whereas oncogene induction in differentiated cells induces massive DNA damage, mammary stem cells are resistant, owing to a preemptive program driven by the transcription factor ZEB1 and the methionine sulfoxide reductase MSRB3. The prevention of oncogene-induced DNA damage precludes induction of the oncosuppressive p53-dependent DNA-damage response, thereby increasing stem cells' intrinsic susceptibility to malignant transformation. In accord with this model, a subclass of breast neoplasms exhibit unique pathological features, including high ZEB1 expression, a low frequency of TP53 mutations and low CIN.

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Figure 1: Cellular hierarchy of the normal human mammary gland.
Figure 2: Mammary stem cells exhibit low levels of DNA damage after RAS transduction.
Figure 3: Stemness-related expression of ZEB1 prevents oncogene-induced DNA damage in normal and cancer cells.
Figure 4: The methionine sulfoxide reductase MSRB3 is required for ZEB1-dependent protection against oncogene-induced DNA damage.
Figure 5: The ZEB1–MSRB3 axis is associated with low CNA in breast cancer.
Figure 6: ZEB1 expression fosters malignant transformation and prevents the onset of chromosomal instability.

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The authors thank I. Treilleux and G. Clapisson (Resource Biological Center) for providing tumor samples; N. Nazaret for sequencing analysis; P. Kannouche for providing help and laboratory space for the DNA-fiber spreading analysis; I. Iacono, S. Bréjon, A. Pierrot, J. Perrossier and S. Croze for technical support; D. Nègre (CIRI/EVIR) for providing the pWPIR-GFP lentiviral vector; C. Eaves (Terry Fox Laboratory) for providing cells; and M. Tommasino (IARC) for providing vectors. We thank the CRCM animal core facility for animal housing and the CRCM flow cytometry core (M.L. Thibullt) for technical assistance. We also thank B. Manship for critical reading of the manuscript. This work was supported by funding from the Ligue Nationale contre le Cancer (EL2011.LNCC/AP and EL2016.LNCC/AIP) and the Project National Cancer Association, Lyon Integrated Research Institute in Cancer (LYRIC, INCa 4664) to A.P. This work was additionally supported by funding from the Institut National contre le Cancer (project PLBio 2015-266) to P.S., the Institut National contre le Cancer (project PLBIO12-007) to E.D., a Cancer-DGOS grant (TRANSLA11-103) to E.C.-J. and the Marseille Integrated Research Institute in Cancer (INCa 6038) to D.B.

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A.-P.M. coordinated all experiments performed on cell lines and assisted in data interpretation and preparation of figures and methods. C.C., C.P. and F.F. produced and characterized all HMEC-derived cell lines. R.M.P., E.R., P.S., E.T. and C.M.-L. performed bioinformatics and statistical analyses, and R.M.P. assisted in preparation of figures and methods. M.D.-S., N.R.-R. and F.P.-L. performed tumor analyses. V.C. coordinated aCGH analyses. J.L. and Q.W. coordinated sequencing analyses. A.W. and F.B. performed gene expression analysis. E.D. and A.T. performed DNA-fiber spreading analysis, and A.T. assisted in preparation of figures and methods. J.-L.J. provided normal breast samples. E.C.-J., C.G. and J.W. performed normal breast tissue isolation. E.C.-J., C.G. and D.B. conceived experiments on normal human mammary glands. O.C. carried out the in vivo experiments. P.F. isolated mRNA from normal mammary glands. J.G. and A.M.V. performed nucleotide quantification experiments. J.C., S.A., A.T., F.H. and A.M.V. assisted in data interpretation. D.G.C. provided critical evaluation of the manuscript and scientific content. A.P. conceived the project, designed experiments, interpreted data and wrote the manuscript. All authors read and approved the final version of the manuscript.

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Correspondence to Alain Puisieux.

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The authors declare no competing financial interests.

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Morel, AP., Ginestier, C., Pommier, R. et al. A stemness-related ZEB1–MSRB3 axis governs cellular pliancy and breast cancer genome stability. Nat Med 23, 568–578 (2017).

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