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Transient PLK4 overexpression accelerates tumorigenesis in p53-deficient epidermis

Nature Cell Biology volume 18, pages 100–110 (2016)Cite this article

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Abstract

Aneuploidy is found in most solid tumours, but it remains unclear whether it is the cause or the consequence of tumorigenesis. Using Plk4 overexpression (PLK4OE) during epidermal development, we assess the impact of centrosome amplification and aneuploidy on skin development and tumorigenesis. PLK4OE in the developing epidermis induced centrosome amplification and multipolar divisions, leading to p53 stabilization and apoptosis of epidermal progenitors. The resulting delayed epidermal stratification led to skin barrier defects. Plk4 transgene expression was shut down postnatally in the surviving mice and PLK4OE mice never developed skin tumours. Concomitant PLK4OE and p53 deletion (PLK4OE/p53cKO) rescued the differentiation defects, but did not prevent the apoptosis of PLK4OE cells. Remarkably, the short-term presence of cells with supernumerary centrosomes in PLK4OE/p53cKO mice was sufficient to generate aneuploidy in the adult epidermis and triggered spontaneous skin cancers with complete penetrance. These results reveal that aneuploidy induced by transient centrosome amplification can accelerate tumorigenesis in p53-deficient cells.

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Figure 1: PLK4 overexpression impairs skin stratification.
Figure 2: PLK4 overexpression induced epidermal progenitor apoptosis.
Figure 3: p53 deletion rescues the defect of skin stratification in PLK4OE mice.
Figure 4: PLK4 overexpression induced p53-dependent and-independent apoptosis.
Figure 5: PLK4 overexpression causes aneuploidy only in p53-deficient progenitors.
Figure 6: PLK4OE in p53-deficient progenitors accelerates skin tumour formation.
Figure 7: Model of skin tumorigenesis mediated by PLK4OE in p53-deficient epidermis.

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  • 10 December 2015

    In the version of this Article originally published online, Bram Boeckx and Diether Lambrechts should have had a second affiliation: Vesalius Research Center, VIB, 3000 Leuven, Belgium. This is now correct in all versions of the Article.

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Acknowledgements

C.B. is an investigator of WELBIO; Ö.S. is supported by a TELEVIE fellowship. This work was supported by the FNRS, TELEVIE, the PAI programme, a research grant from the Fondation contre le Cancer, the ULB Fondation, the foundation Bettencourt Schueller, the foundation Baillet Latour, a grant from the European Research Council (ERC) and a grant from AICR now worldwide cancer research (13-0170) to V.M. and R.B.

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Authors and Affiliations

  1. Université Libre de Bruxelles, IRIBHM, Brussels B-1070, Belgium

    Özdemirhan Serçin, Jean-Christophe Larsimont, Andrea E. Karambelas, Virginie Moers & Cédric Blanpain

  2. Institut Curie, Centre de Recherche, Paris F-75248, France

    Veronique Marthiens & Renata Basto

  3. Department of Oncology, Laboratory for Translational Genetics, KU Leuven 3000, Belgium

    Bram Boeckx & Diether Lambrechts

  4. Vesalius Research Center, VIB, 3000 Leuven, Belgium

    Bram Boeckx & Diether Lambrechts

  5. Department of Pathology, Erasme Hospital, Université Libre de Bruxelles, Brussels B-1070, Belgium

    Marie Le Mercier

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Contributions

Ö.S. and C.B. designed the experiments and performed data analysis; Ö.S. performed most of the experiments, J.-C.L. and A.E.K. conducted proliferation experiments; V.M. performed RNA purification; M.L.M. performed FISH experiments; D.L. and B.B. performed tumour sequencing and analysis. V.Ma. and R.B. generated the PLK4OE mice; Ö.S. and C.B. wrote the manuscript.

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Correspondence to Cédric Blanpain.

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Integrated supplementary information

Supplementary Figure 1 Schematic representation of transgene recombination.

(a) A loxp-STOP-loxp cassette prevents expression of mCherry-Plk4 construct. (b) Upon initiation of K14 expression in mouse skin epidermis, Cre accumulates and recombines loxP sites and enables Plk4 overexpression in mice older than E14. Unrecombined and recombined specific primers and PCR are detailed in Methods.

Supplementary Figure 2 mCherry-Plk4 localizes to centrosomes in mouse skin epidermis and causes centrosome supernumerary.

(a) Keratinocytes extracted from E16.5 embryos were stained for centrosome marker γ-tubulin. mCherry-Plk4 co-localizes with γ-tubulin, insets show higher magnification of the same image for co-localization. (b) Quantification of percentage of cells mCherry-positive and cells with >2 centrosomes. Data represent the mean of n = 3 mice. (≥30 cells per mouse were counted) Nuclei were stained with DAPI and white scale bars represent 5 μm. Source data are available in Supplementary Table 1.

Supplementary Figure 3 Plk4 overexpression causes skin development defects.

(a) Newborn PLK4OE mice show shiny skin and open eyes. (b) Close up image of (a) reveals incomplete eyelid formation in PLK4OE but not control mice. (c,d) Macroscopic image of newborn mice show that only PLK4OE but not control mice exhibit shiny face (c) and back skin (d).

Supplementary Figure 4 PCR of recombined and unrecombined Plk4 transgene.

(a) Recombination of the transgene was detected by genomic PCR only in double transgenic mouse at E16.5, P1, adult and in different tumours but not in control mice or CRE negative mice, non-recombined allele was detected by a primer designed specifically to amplify the cells which did not recombine the transgene. ß-actin was used as a loading control.

Supplementary Figure 5 Expression of mCherry-PLK4 mRNA is lost in adult mice.

(a) Expression of mCherry-Plk4 was tested by RT-PCR in P1 and adult mice. Only P1 but not adult PLK4OE and PLK4OE/p53cKO mice show amplification of the RT product. ß-actin was used as a loading control.

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Serçin, Ö., Larsimont, JC., Karambelas, A. et al. Transient PLK4 overexpression accelerates tumorigenesis in p53-deficient epidermis. Nat Cell Biol 18, 100–110 (2016). https://doi.org/10.1038/ncb3270

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