p63 regulates commitment to the prostate cell lineage

Sabina Signoretti

Maira Pires

Meghan Lindauer

James Horner

Philip Kantoff+2 authors

^{Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115;}^{Department of Medical Oncology, Dana–Farber Cancer Institute, Harvard Medical School, Boston, MA 02115;}^{Department of Cell Biology, Harvard Medical School, Boston, MA 02115; and}^{Broad Institute, Massachusetts Institute of Technology, Harvard University, and the Whitehead Institute for Biomedical Research, Cambridge, MA 02141}

^{To whom correspondence may be addressed. E-mail:}gro.srentrap@itterongiss or ude.dravrah.icfd@adol_omissam.

Edited by Owen N. Witte, University of California, Los Angeles, CA, and approved June 27, 2005

Received 2005 Mar 9

Abstract

Molecular mechanisms underlying prostate and urothelial development remain unclear. This situation presents major limitations in identifying the cell type(s) and molecular events involved in the development of prostate and bladder cancer. It has been shown that mice lacking the basal cell marker p63 present several epithelial defects, including epidermis and prostate buds agenesis and urothelial abnormalities. Here, we use the p63–/– mouse as a tool to define cell lineages in the prostate epithelium and urothelium. By complementing p63–/– blastocysts with p63+/+ β-galactosidase (β-gal)-positive ES cells, we show that secretory cells of the prostate originate from p63-positive basal progenitor cells. Importantly, our urogenital sinus transplantation studies demonstrate that p63 prevents intestinal differentiation of the urogenital sinus endoderm and is therefore required to maintain commitment to the prostate cell lineage. Finally, in contrast with the prostate findings, analysis of the urothelium from rescued p63–/– chimeras shows that umbrella (superficial) cells can develop and be maintained independently from p63-positive basal and intermediate cells.

Keywords: animal model, development, stem cell, urothelium, mouse model

Abstract

Unraveling the means by which the development and renewal of normal tissues occurs is an essential step for the elucidation of the mechanisms underlying the development of pathological processes, including cancer. Indeed, the discovery of hematopoietic stem cells has led to considerable progress toward the identification of the cell types and the molecular pathways implicated in the development of hematopoietic malignancies (1, 2). Although epithelial stem cells have been identified in both the skin and intestine, how the prostate epithelium and the urothelium are formed and maintained is unknown, and the existence of stem cells in these epithelia is controversial. It has been proposed that, in both the prostate epithelium and the urothelium, the basal cells represent or include the progenitor/stem cells (3–6). However, because both prostate secretory cells and urothelial umbrella (superficial) cells have the ability to divide, they may represent independent cell lineages with self-renewal capacities (7–10). Thus, the clarification of the developmental hierarchy among the different epithelial cell types that constitute the prostate and the urothelium is crucial for the future identification of the cell of origin of genitourinary malignancies, including prostate and bladder cancer.

It has been shown that p63 is a marker of basal cells (11) and is required for normal development of several epithelial tissues, including squamous epithelia, urothelium, and mammary, salivary, lacrymal, and prostate glands (12–14). To investigate whether p63-positive basal cells are the progenitor/stem cells of the prostate epithelium and urothelium, we rescued the developmental abnormalities of the p63–/– mice by injecting p63+/+ ROSA26 (β-gal-positive) ES cells into blastocysts isolated from p63+/– crosses. Information regarding cell lineages in the prostate epithelium and urothelium were obtained by assessing the contribution of the p63–/– cells to these epithelia. In addition, postnatal development of p63–/– urogenital sinus (UGS) was addressed by transplantation studies as well.

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Acknowledgments

We thank Stuart H. Orkin, David M. Livingston, Myles Brown, and Ramesh A. Shivdasani for critical reading of the manuscript. This work was supported by grants from the Department of Defense and the National Cancer Institute [Specialized Program of Research Excellence (SPORE) in RenalCancer], the Hershey Prostate Cancer/Survivor Walk Award, CaPCURE Award, and an award from Istituto Dermopatico dell'Immacolata (Rome) (to S.S.); and grants from the National Cancer Institute (R01, P01, and Specialized Program of Research Excellence in Prostate Cancer) and the Gelb Center for genitourinary oncology at the Dana–Farber Cancer Institute, a Novartis Investigator grant, and a CaPCURE Award (to M.L.).

Acknowledgments

Notes

Author contributions: S.S., P.W.K., W.R.S., and M. Loda designed research; S.S., M.M.P., M. Lindauer, J.W.H., C.G., S.D., and P.M. performed research; F.M. contributed new reagents/analytic tools; S.S., C.G., and M. Loda analyzed data; and S.S. wrote the paper.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviation: UGS, urogenital sinus.

Notes

This paper was submitted directly (Track II) to the PNAS office.

Abbreviation: UGS, urogenital sinus.

References

1. Sawyers, C. L., Denny, C. T. & Witte, O. N. (1991) Cell64, 337–350. [[PubMed]
2. Passegue, E., Jamieson, C. H., Ailles, L. E. & Weissman, I. L. (2003) Proc. Natl. Acad. Sci. USA100, Suppl. 1, 11842–11849.
3. De Marzo, A. M., Nelson, W. G., Meeker, A. K. & Coffey, D. S. (1998) J. Urol.160, 2381–2392. [[PubMed]
4. Wang, Y., Hayward, S., Cao, M., Thayer, K. & Cunha, G. (2001) Differentiation (Berlin)68, 270–279. [[PubMed]
5. Marceau, N(1990) Lab. Invest.63, 4–20. [[PubMed][Google Scholar]
6. Bonkhoff, H. & Remberger, K. (1996) Prostate28, 98–106. [[PubMed]
7. Evans, G. S. & Chandler, J. A. (1987) Prostate11, 339–351. [[PubMed]
8. English, H. F., Santen, R. J. & Isaacs, J. T. (1987) Prostate11, 229–242. [[PubMed]
9. Messier, B. & Leblond, C. P. (1960) Am. J. Anat.106, 247–285. [[PubMed]
10. Tsujimura, A., Koikawa, Y., Salm, S., Takao, T., Coetzee, S., Moscatelli, D., Shapiro, E., Lepor, H., Sun, T. T. & Wilson, E. L. (2002) J. Cell Biol.157, 1257–1265.
11. Yang, A., Kaghad, M., Wang, Y., Gillett, E., Fleming, M. D., Dotsch, V., Andrews, N. C., Caput, D. & McKeon, F. (1998) Mol. Cell2, 305–316. [[PubMed]
12. Yang, A., Schweitzer, R., Sun, D., Kaghad, M., Walker, N., Bronson, R. T., Tabin, C., Sharpe, A., Caput, D., Crum, C. & McKeon, F. (1999) Nature398, 714–718. [[PubMed]
13. Signoretti, S., Waltregny, D., Dilks, J., Isaac, B., Lin, D., Garraway, L., Yang, A., Montironi, R., McKeon, F. & Loda, M. (2000) Am. J. Pathol.157, 1769–1775.
14. Mills, A. A., Zheng, B., Wang, X. J., Vogel, H., Roop, D. R. & Bradley, A. (1999) Nature398, 708–713. [[PubMed]
15. Gounari, F., Signoretti, S., Bronson, R., Klein, L., Sellers, W. R., Kum, J., Siermann, A., Taketo, M. M., von Boehmer, H. & Khazaie, K. (2002) Oncogene21, 4099–4107. [[PubMed]
16. van Bokhoven, H. & McKeon, F. (2002) Trends Mol. Med.8, 133–139. [[PubMed]
17. Kurita, T., Medina, R. T., Mills, A. A. & Cunha, G. R. (2004) Development (Cambridge, U.K.)131, 4955–4964. [[PubMed]
18. Bhatia-Gaur, R., Donjacour, A. A., Sciavolino, P. J., Kim, M., Desai, N., Young, P., Norton, C. R., Gridley, T., Cardiff, R. D., Cunha, G. R., et al. (1999) Genes Dev.13, 966–977.
19. Bry, L., Falk, P., Huttner, K., Ouellette, A., Midtvedt, T. & Gordon, J. I. (1994) Proc. Natl. Acad. Sci. USA91, 10335–10339.
20. Suh, E. & Traber, P. G. (1996) Mol. Cell. Biol.16, 619–625.
21. Moskaluk, C. A., Zhang, H., Powell, S. M., Cerilli, L. A., Hampton, G. M. & Frierson, H. F., Jr. (2003) Mod. Pathol.16, 913–919. [[PubMed]
22. Moll, R., Wu, X. R., Lin, J. H. & Sun, T. T. (1995) Am. J. Pathol.147, 1383–1397.
23. Urist, M. J., Di Como, C. J., Lu, M. L., Charytonowicz, E., Verbel, D., Crum, C. P., Ince, T. A., McKeon, F. D. & Cordon-Cardo, C. (2002) Am. J. Pathol.161, 1199–1206.
24. Erman, A., Jezernik, K., Stiblar-Martincic, D., Romih, R. & Veranic, P. (2001) Histochem. Cell Biol.115, 309–316. [[PubMed]
25. Ayres, P. H., Shinohara, Y. & Frith, C. H. (1985) J. Urol.133, 506–512. [[PubMed]
26. Jost, S. P. (1985) J. Anat.143, 39–43.
27. Mackillop, W. J., Bizarri, J. P. & Ward, G. K. (1985) Cancer Res.45, 4360–4365. [[PubMed]
28. Varley, C. L., Stahlschmidt, J., Smith, B., Stower, M. & Southgate, J. (2004) Am. J. Pathol.164, 1789–1798.