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SMAD4-dependent barrier constrains prostate cancer growth and metastatic progression

Nature volume 470, pages 269–273 (2011)Cite this article

Abstract

Effective clinical management of prostate cancer (PCA) has been challenged by significant intratumoural heterogeneity on the genomic and pathological levels and limited understanding of the genetic elements governing disease progression1. Here, we exploited the experimental merits of the mouse to test the hypothesis that pathways constraining progression might be activated in indolent Pten-null mouse prostate tumours and that inactivation of such progression barriers in mice would engender a metastasis-prone condition. Comparative transcriptomic and canonical pathway analyses, followed by biochemical confirmation, of normal prostate epithelium versus poorly progressive Pten-null prostate cancers revealed robust activation of the TGFβ/BMP–SMAD4 signalling axis. The functional relevance of SMAD4 was further supported by emergence of invasive, metastatic and lethal prostate cancers with 100% penetrance upon genetic deletion of Smad4 in the Pten-null mouse prostate. Pathological and molecular analysis as well as transcriptomic knowledge-based pathway profiling of emerging tumours identified cell proliferation and invasion as two cardinal tumour biological features in the metastatic Smad4/Pten-null PCA model. Follow-on pathological and functional assessment confirmed cyclin D1 and SPP1 as key mediators of these biological processes, which together with PTEN and SMAD4, form a four-gene signature that is prognostic of prostate-specific antigen (PSA) biochemical recurrence and lethal metastasis in human PCA. This model-informed progression analysis, together with genetic, functional and translational studies, establishes SMAD4 as a key regulator of PCA progression in mice and humans.

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Figure 1: SMAD4 is a putative suppressor of prostate tumour progression.
Figure 2: Smad4 deletion drives progression of Pten -deficient prostate tumour to highly aggressive prostate cancer metastatic to lymph node and lung.
Figure 3: Ccnd1 and Spp1 are mediators of prostate tumour cell proliferation and metastasis.
Figure 4: Prognostic potential of a four-gene signature in human PCA.

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Gene Expression Omnibus

Data deposits

The microarray data have been deposited in the GOE database with accession number GSE25140.

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Acknowledgements

The authors are grateful to the late W. Gerald for providing the primary gene expression data and clinical outcome files13. We thank S. Zhou for excellent mouse husbandry and care, B. Xiong and G. Tonon for bioinformatic assistance, and S. Jia, J. M. Stommel, J. Paik, M. Kim and A. C. Kimmelman for helpful discussion. We thank M. Vidal, the Ellison Foundation and DFCI ISR for support of ORF cloning efforts, R. Maser for MSCV-puro-v5 gateway vector, W. Hahn for shRNA constructs. We thank the DF/HCC Specialized Histopathology Core and the DF/HCC Tissue Microarray and Imaging core for the TMA IHC staining; the DFCI/BWH Center for Molecular Oncologic Pathology (CMOP) for the quantification of the IHC. Z.D was supported by the Damon Runyon Cancer Research Foundation. D.H. was supported by a graduate fellowship from the National Science Foundation. H.Z. was supported by the Helen Hay Whitney Foundation. Y.A.W. was supported by the Multiple Myeloma Research Foundation. This work is supported by the Belfer Institute for Applied Cancer Science, NCI U01-CA84313 (L.C. and R.A.D.), DF/HCC SPORE in Prostate Cancer P50 CA090381-08 (Z.D.), the National Cancer Institute (M.L. RO1CA131945 and P50 CA90381, L.M. RO1 5R01CA136578, M.S. R01CA141298), and the Linda and Arthur Gelb Center for Translational Research (M.L.). R.A.D. was supported by an American Cancer Society Research Professorship and L.M. was supported by the Prostate Cancer Foundation.

Author information

Author notes

  1. Chang-Jiun Wu and Gerald C. Chu: These authors contributed equally to this work.

Authors and Affiliations

  1. Belfer Institute for Applied Cancer Science, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Zhihu Ding, Chang-Jiun Wu, Gerald C. Chu, Yonghong Xiao, Dennis Ho, Samuel R. Perry, Emma S. Labrot, Baoli Hu, Shan Jiang, Hongwu Zheng, Alexander H. Stegh, Kenneth L. Scott, Y. Alan Wang, Lynda Chin & Ronald A. DePinho

  2. Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Zhihu Ding, Chang-Jiun Wu, Gerald C. Chu, Yonghong Xiao, Dennis Ho, Samuel R. Perry, Emma S. Labrot, Xiaoqiu Wu, Rosina Lis, Baoli Hu, Shan Jiang, Hongwu Zheng, Alexander H. Stegh, Kenneth L. Scott, Y. Alan Wang, Massimo Loda, Lynda Chin & Ronald A. DePinho

  3. Department of Genetics, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Zhihu Ding, Chang-Jiun Wu, Dennis Ho, Hongwu Zheng, Alexander H. Stegh, Kenneth L. Scott, David E. Hill, Lynda Chin & Ronald A. DePinho

  4. Department of Medicine, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Zhihu Ding, Chang-Jiun Wu, Dennis Ho, Hongwu Zheng, Alexander H. Stegh, Kenneth L. Scott, Lynda Chin & Ronald A. DePinho

  5. Department of Pathology, Brigham and Women’s Hospital, Boston, 02115, Massachusetts, USA

    Gerald C. Chu & Massimo Loda

  6. McArdle Lab for Cancer Research, University of Wisconsin, Madison, 53706-1526, Wisconsin, USA

    Jingfang Zhang

  7. Center for Molecular Oncologic Pathology, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Xiaoqiu Wu, Rosina Lis & Massimo Loda

  8. Pediatric Oncology, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Yujin Hoshida & Todd R. Golub

  9. The Eli and Edythe L. Broad Institute, Massachusetts Institute of Technology and Harvard University, Cambridge, 02142, Massachusetts, USA

    Yujin Hoshida & Todd R. Golub

  10. Department of Statistics, Stanford University, Stanford, 94305, California, USA

    David Hiller & Wing H. Wong

  11. Renal Cancer Program, Dana-Farber/Harvard Cancer Center, Boston, 02115, Massachusetts, USA

    Sabina Signoretti

  12. Department of Medicine, Massachusetts General Hospital Cancer Center, Boston, 02114, Massachusetts, USA

    Nabeel Bardeesy

  13. Center for Cancer Systems Biology and Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    David E. Hill

  14. Department of Dermatology, Brigham and Women’s Hospital, Boston, 02115, Massachusetts, USA

    Lynda Chin

  15. Department of Epidemiology, Harvard School of Public Health, Boston, 02115, Massachusetts, USA

    Meir J. Stampfer & Lorelei Mucci

  16. Department of Nutrition, Harvard School of Public Health, Boston, 02115, Massachusetts, USA

    Meir J. Stampfer

  17. Channing Laboratory, Brigham and Women’s Hospital, Boston, 02115, Massachusetts, USA

    Meir J. Stampfer & Lorelei Mucci

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  1. Zhihu Ding
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Contributions

Z.D. designed and performed the experiments. L.C. and R.A.D. supervised experiments and computational analysis and contributed as senior authors. C.J.W., Y.X., Y.H., D.H., T.R.G., M.J.S., W.H.W. and L.M. performed the computational analysis. G.C.C. provided pathology analyses. X.W., R.L., S.S. and M.L. performed TMA staining and quantification. N.B. generated Smad4L mouse allele. D.E.H. provided the human ORFeome clones. D.H., J.Z., S.R.P., E.S.L., B.H., S.J., H.Z., A.H.S. and K.L.S. performed the experiments. Y.A.W. contributed to the writing of the manuscript.

Corresponding authors

Correspondence to Lynda Chin or Ronald A. DePinho.

Ethics declarations

Competing interests

The gene signature and technology developed in this paper have been licensed by Metamark GENETICS (http://www.metamarkgenetics.com/). L.C. and R.A.D. are the founders of Metamark GENETICS. Z.D., C.W. and M.L. are consultants for Metamark GENETICS.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-21 with legends and Supplementary Tables 1-8. (PDF 1558 kb)

Supplementary Data 1

This table shows a complete differentially expressed gene list was generated by a class comparison between WT (n=3) and Ptenpc-/- (n=5) anterior prostates at 15 weeks of age. (XLS 705 kb)

Supplementary Data 2

This table shows a complete differentially expressed gene list was generated by a class comparison between Ptenpc-/- (n=5) and Ptenpc-/-Smad4pc-/- (n=5) anterior prostate tumors at 15 weeks of age. (XLS 103 kb)

Supplementary Data 3

This table shows a complete differentially expressed gene list was generated by a class comparison between Ptenpc-/- and Ptenpc-/-p53pc-/- mouse anterior prostates at 15 weeks of age. (XLS 89 kb)

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Ding, Z., Wu, CJ., Chu, G. et al. SMAD4-dependent barrier constrains prostate cancer growth and metastatic progression. Nature 470, 269–273 (2011). https://doi.org/10.1038/nature09677

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