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Hedgehog/Wnt feedback supports regenerative proliferation of epithelial stem cells in bladder


Epithelial integrity in metazoan organs is maintained through the regulated proliferation and differentiation of organ-specific stem and progenitor cells. Although the epithelia of organs such as the intestine regenerate constantly and thus remain continuously proliferative1, other organs, such as the mammalian urinary bladder, shift from near-quiescence to a highly proliferative state in response to epithelial injury2,3,4. The cellular and molecular mechanisms underlying this injury-induced mode of regenerative response are poorly defined. Here we show in mice that the proliferative response to bacterial infection or chemical injury within the bladder is regulated by signal feedback between basal cells of the urothelium and the stromal cells that underlie them. We demonstrate that these basal cells include stem cells capable of regenerating all cell types within the urothelium, and are marked by expression of the secreted protein signal Sonic hedgehog (Shh). On injury, Shh expression in these basal cells increases and elicits increased stromal expression of Wnt protein signals, which in turn stimulate the proliferation of both urothelial and stromal cells. The heightened activity of this signal feedback circuit and the associated increase in cell proliferation appear to be required for restoration of urothelial function and, in the case of bacterial injury, may help clear and prevent further spread of infection. Our findings provide a conceptual framework for injury-induced epithelial regeneration in endodermal organs, and may provide a basis for understanding the roles of signalling pathways in cancer growth and metastasis.

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Figure 1: Injury-induced proliferation and Hedgehog signalling in the bladder.
Figure 2: Shh-expressing basal cells repopulate the urothelium and form organoids in vitro.
Figure 3: Gli1 mediates injury-induced proliferation, restoration of urothelial integrity and reduction of infectious spread.
Figure 4: Hedgehog-induced expression of stromal Wnt signals mediates urothelial and stromal proliferation.


  1. 1

    van der Flier, L. G. & Clevers, H. Stem cells, self-renewal, and differentiation in the intestinal epithelium. Annu. Rev. Physiol. 71, 241–260 (2009)

    CAS  Article  Google Scholar 

  2. 2

    Hung, C. S., Dodson, K. W. & Hultgren, S. J. A murine model of urinary tract infection. Nature Protocols 4, 1230–1243 (2009)

    CAS  Article  Google Scholar 

  3. 3

    Mysorekar, I. U. & Hultgren, S. J. Mechanisms of uropathogenic Escherichia coli persistence and eradication from the urinary tract. Proc. Natl Acad. Sci. USA 103, 14170–14175 (2006)

    ADS  CAS  Article  Google Scholar 

  4. 4

    Mysorekar, I. U., Isaacson-Schmid, M., Walker, J. N., Mills, J. C. & Hultgren, S. J. Bone morphogenetic protein 4 signaling regulates epithelial renewal in the urinary tract in response to uropathogenic infection. Cell Host Microbe 5, 463–475 (2009)

    CAS  Article  Google Scholar 

  5. 5

    Hicks, R. M. The mammalian urinary bladder: an accommodating organ. Biol. Rev. Camb. Philos. Soc. 50, 215–246 (1975)

    CAS  Article  Google Scholar 

  6. 6

    Kurzrock, E. A., Lieu, D. K., Degraffenried, L. A., Chan, C. W. & Isseroff, R. R. Label-retaining cells of the bladder: candidate urothelial stem cells. Am. J. Physiol. Renal Physiol. 294, F1415–F1421 (2008)

    CAS  Article  Google Scholar 

  7. 7

    Hooton, T. M. & Stamm, W. E. Diagnosis and treatment of uncomplicated urinary tract infection. Infect. Dis. Clin. North Am. 11, 551–581 (1997)

    CAS  Article  Google Scholar 

  8. 8

    Klumpp, D. J. et al. Uropathogenic Escherichia coli induces extrinsic and intrinsic cascades to initiate urothelial apoptosis. Infect. Immun. 74, 5106–5113 (2006)

    CAS  Article  Google Scholar 

  9. 9

    Podlasek, C. A., Barnett, D. H., Clemens, J. Q., Bak, P. M. & Bushman, W. Prostate development requires Sonic hedgehog expressed by the urogenital sinus epithelium. Dev. Biol. 209, 28–39 (1999)

    CAS  Article  Google Scholar 

  10. 10

    Haraguchi, R. et al. Molecular analysis of coordinated bladder and urogenital organ formation by Hedgehog signaling. Development 134, 525–533 (2007)

    CAS  Article  Google Scholar 

  11. 11

    Karhadkar, S. S. et al. Hedgehog signalling in prostate regeneration, neoplasia and metastasis. Nature 431, 707–712 (2004)

    ADS  CAS  Article  Google Scholar 

  12. 12

    Bai, C. B., Auerbach, W., Lee, J. S., Stephen, D. & Joyner, A. L. Gli2, but not Gli1, is required for initial Shh signaling and ectopic activation of the Shh pathway. Development 129, 4753–4761 (2002)

    CAS  Article  Google Scholar 

  13. 13

    Lawson, D. A., Xin, L., Lukacs, R. U., Cheng, D. & Witte, O. N. Isolation and functional characterization of murine prostate stem cells. Proc. Natl Acad. Sci. USA 104, 181–186 (2007)

    ADS  CAS  Article  Google Scholar 

  14. 14

    Harfe, B. D. et al. Evidence for an expansion-based temporal Shh gradient in specifying vertebrate digit identities. Cell 118, 517–528 (2004)

    CAS  Article  Google Scholar 

  15. 15

    Muzumdar, M. D., Tasic, B., Miyamichi, K., Li, L. & Luo, L. A global double-fluorescent Cre reporter mouse. Genesis 45, 593–605 (2007)

    CAS  Article  Google Scholar 

  16. 16

    Wu, X. R. & Sun, T. T. Molecular cloning of a 47 kDa tissue-specific and differentiation-dependent urothelial cell surface glycoprotein. J. Cell Sci. 106, 31–43 (1993)

    CAS  PubMed  Google Scholar 

  17. 17

    Ahn, S. & Joyner, A. L. Dynamic changes in the response of cells to positive Hedgehog signaling during mouse limb patterning. Cell 118, 505–516 (2004)

    CAS  Article  Google Scholar 

  18. 18

    Kimura, H., Stephen, D., Joyner, A. & Curran, T. Gli1 is important for medulloblastoma formation in Ptc1+/− mice. Oncogene 24, 4026–4036 (2005)

    CAS  Article  Google Scholar 

  19. 19

    Hagberg, L. et al. Ascending, unobstructed urinary tract infection in mice caused by pyelonephritogenic Escherichia coli of human origin. Infect. Immun. 40, 273–283 (1983)

    CAS  PubMed  PubMed Central  Google Scholar 

  20. 20

    Goessling, W. et al. Genetic interaction of PGE2 and Wnt signaling regulates developmental specification of stem cells and regeneration. Cell 136, 1136–1147 (2009)

    CAS  Article  Google Scholar 

  21. 21

    Lustig, B. et al. Negative feedback loop of Wnt signaling through upregulation of conductin/axin2 in colorectal and liver tumors. Mol. Cell. Biol. 22, 1184–1193 (2002)

    CAS  Article  Google Scholar 

  22. 22

    Klein, P. S. & Melton, D. A. A molecular mechanism for the effect of lithium on development. Proc. Natl Acad. Sci. USA 93, 8455–8459 (1996)

    ADS  CAS  Article  Google Scholar 

  23. 23

    Su, L. K. et al. Multiple intestinal neoplasia caused by a mutation in the murine homolog of the APC gene. Science 256, 668–670 (1992)

    ADS  CAS  Article  Google Scholar 

  24. 24

    Fendrich, V. et al. Hedgehog signaling is required for effective regeneration of exocrine pancreas. Gastroenterology 135, 621–631 (2008)

    CAS  Article  Google Scholar 

  25. 25

    Nicolle, L. E. Uncomplicated urinary tract infection in adults including uncomplicated pyelonephritis. Urol. Clin. North Am. 35, 1–12 (2008)

    Article  Google Scholar 

  26. 26

    Foxman, B. Recurring urinary tract infection: incidence and risk factors. Am. J. Public Health 80, 331–333 (1990)

    CAS  Article  Google Scholar 

  27. 27

    Yauch, R. L. et al. A paracrine requirement for hedgehog signalling in cancer. Nature 455, 406–410 (2008)

    ADS  CAS  Article  Google Scholar 

  28. 28

    Tian, H. et al. Hedgehog signaling is restricted to the stromal compartment during pancreatic carcinogenesis. Proc. Natl Acad. Sci. USA 106, 4254–4259 (2009)

    ADS  CAS  Article  Google Scholar 

  29. 29

    Bubendorf, L. et al. Metastatic patterns of prostate cancer: an autopsy study of 1,589 patients. Hum. Pathol. 31, 578–583 (2000)

    CAS  Article  Google Scholar 

  30. 30

    Wallmeroth, A. et al. Patterns of metastasis in muscle-invasive bladder cancer (pT2–4): an autopsy study on 367 patients. Urol. Int. 62, 69–75 (1999)

    CAS  Article  Google Scholar 

  31. 31

    Kurzrock, E. A., Lieu, D. K., deGraffenried, L. A. & Isseroff, R. R. Rat urothelium: improved techniques for serial cultivation, expansion, freezing and reconstitution onto acellular matrix. J. Urol. 173, 281–285 (2005)

    Article  Google Scholar 

  32. 32

    Ibla, J. C. & Khoury, J. Methods to assess tissue permeability. Methods Mol. Biol. 341, 111–117 (2006)

    PubMed  Google Scholar 

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We thank A. Oro and J. Brooks for their critical reading of the manuscript, and the Stanford Center for Digestive Diseases for help with laser capture microdissection. This research was supported in part by grants from the Department of Defense and from the National Institutes of Health (P.A.B.) and a Pathway to Independence Award (K99/R00) to I.U.M. P.A.B. is an investigator of the Howard Hughes Medical Institute.

Author information




K.S. and P.A.B. conceived ideas and experimental design. K.S. performed the experiments. N.G. aided in immunohistochemical analysis, J.L. and J.K. helped with mouse strains, A.L. assisted with in vitro cell culture studies, L.Q. performed the genotyping of experimental mice, and I.U.M. helped analyse data. K.S. and P.A.B wrote the manuscript.

Corresponding authors

Correspondence to Kunyoo Shin or Philip A. Beachy.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-19 with legends and Supplementary Tables 1-6. (PDF 25818 kb)

Supplementary Movie 1

The movie shows an organoid grown by culturing FACS-isolated Shh-expressing cells for 7 weeks in Matrigel was imaged and a portion was reconstructed from approximately ••• of the series of confocal EGFP images (Supplementary Fig. 11g,i). This reconstruction was rotated to display the shape and lumen of the organoid. Bladder cells were isolated directly from TM-injected ShhCreER/WT; R26mTmG/WT mice (Imaris software, Bitplane Scientific Software). (MPG 3172 kb)

Supplementary Movie 2

The movie shows organoid grown from a single cell and imaged as in Supplementary Movie 1, except that cells derived from a previously grown organoid (P1 organoid; Supplementary Fig. 11h,j; Imaris software, Bitplane Scientific Software). (MPG 2850 kb)

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Shin, K., Lee, J., Guo, N. et al. Hedgehog/Wnt feedback supports regenerative proliferation of epithelial stem cells in bladder. Nature 472, 110–114 (2011).

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