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Wt1 is required for cardiovascular progenitor cell formation through transcriptional control of Snail and E-cadherin

Nature Genetics volume 42, pages 89–93 (2010)Cite this article

Abstract

The epicardial epithelial-mesenchymal transition (EMT) is hypothesized to generate cardiovascular progenitor cells that differentiate into various cell types, including coronary smooth muscle and endothelial cells, perivascular and cardiac interstitial fibroblasts and cardiomyocytes. Here we show that an epicardial-specific knockout of the gene encoding Wilms' tumor-1 (Wt1) leads to a reduction in mesenchymal progenitor cells and their derivatives. We show that Wt1 is essential for repression of the epithelial phenotype in epicardial cells and during embryonic stem cell differentiation through direct transcriptional regulation of the genes encoding Snail (Snai1) and E-cadherin (Cdh1), two of the major mediators of EMT. Some mesodermal lineages do not form in Wt1-null embryoid bodies, but this effect is rescued by the expression of Snai1, underscoring the importance of EMT in generating these differentiated cells. These new insights into the molecular mechanisms regulating cardiovascular progenitor cells and EMT will shed light on the pathogenesis of heart diseases and may help the development of cell-based therapies.

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Figure 1: Heart defects in epicardial-specific Wt1-mutant embryos.
Figure 2: Wt1 expression is necessary to maintain a mesenchymal phenotype in immortalized epicardial cells.
Figure 3: Wt1 is an activator of Snai1 and a repressor of Cdh1 in epicardial cells.
Figure 4: Wt1 is required for EMT in embryonic stem cells.
Figure 5: Wt1 is required for the formation of some mesodermal lineages in embryoid body differentiation.

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Acknowledgements

We thank A. Cano for the Snai1 promoter (Universidad Autónoma de Madrid, Spain), A. García de Herreros for the antibody to Snail (IMIM-Hospital del Mar, Barcelona, Spain), P. Ruiz-Lozano for the Gata5-Cre transgenic mice (Burnham Institute for Medical Research, La Jolla, California), A. Smith for E14Tg2AIV embryonic stem cells (Wellcome Trust Centre for Stem Cell Research, Cambridge, UK), L. Grotewold for CreERT2-puroR expression construct (University of Edinburgh, UK), H. Morrison for the OPT analysis, F. Kilanowski for assistance with gene targeting, A. Thornburn for help maintaining mouse colonies, C. Nicol for assistance with graphics (Supplementary Fig. 7) and all members of N.D.H.'s laboratory for helpful discussions and comments. This work was supported by the UK Medical Research Council (core grant to N.D.H.) and the Spanish Ministry of Science (grant BFU08-02384 to R.M.-C.). A.E. was supported by EuReGene, an FP6 grant from the European Union (05085). O.M.M.-E. was supported by an EU Marie Curie (FP6) personal fellowship.

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Authors and Affiliations

  1. MRC Human Genetics Unit and the Institute for Genetics and Molecular Medicine, Edinburgh, UK

    Ofelia M Martínez-Estrada, Laura A Lettice, Abdelkader Essafi, Joan Slight, Víctor Velecela, Emma Hall, Judith Reichmann, Paul S Devenney, Peter Hohenstein, Robert E Hill & Nicholas D Hastie

  2. Department of Animal Biology, Faculty of Science, University of Málaga, Málaga, Spain

    Juan Antonio Guadix & Ramón Muñoz-Chapuli

  3. Department of Cancer Stem Cell Biology, Osaka University Graduate School of Medicine, Osaka, Japan

    Naoki Hosen

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  1. Ofelia M Martínez-Estrada
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Contributions

O.M.M.-E. designed, conducted and analyzed experiments and wrote the manuscript. L.A.L. generated Wt1loxP/loxP mice. A.E. conducted part of the promoter and ChIP experiments. J.A.G. conducted immunohistochemistry experiments. J.S. assisted with the embryonic stem experiments. V.V. conducted immunohistochemistry and flow cytometry experiments. E.H. and J.R. conducted initial experiments with immortalized epicardial cells. P.S.D. conducted blastocyst microinjection and helped with mouse maintenance. P.H. helped set up genetic crosses and is responsible for the mouse database. N.H. provided the Wt1GFP/+ knockin mice. R.E.H. contributed to discussion. R.M.-C. designed experiments and cowrote the manuscript. N.D.H. obtained funding, helped with the design and analysis of experiments and cowrote the manuscript.

Corresponding author

Correspondence to Nicholas D Hastie.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7 and Supplementary Table 1 (PDF 2032 kb)

Supplementary Movie 1

3D OPT image of control heart (MOV 4224 kb)

Supplementary Movie 2

3D OPT image of mutant heart (MOV 3505 kb)

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Martínez-Estrada, O., Lettice, L., Essafi, A. et al. Wt1 is required for cardiovascular progenitor cell formation through transcriptional control of Snail and E-cadherin. Nat Genet 42, 89–93 (2010). https://doi.org/10.1038/ng.494

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