Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Uncommitted precursor cells might contribute to increased incidence of embryonal rhabdomyosarcoma in heterozygous Patched1-mutant mice

Oncogene volume 30, pages 4428–4436 (2011)Cite this article

Subjects

Abstract

Embryonal rhabdomyosarcoma (ERMS) is a tumor of the skeletal muscle in children and is frequently initiated by heterozygous germline mutations in the Hedgehog (Hh) receptor Patched1 (Ptch), both in humans and mice. Using a conditional knock-out strategy in Ptchflox/+ mice, we demonstrate that early embryonic stages are more susceptible to ERMS development than later stages and that cells normally not committed to undergo myogenesis at this stage represent the major source of ERMS. We found that deletion of a single copy of the Ptch allele at E9.5 using the ubiquitously active Rosa26CreERT2 resulted in a tumor incidence of 88% but reached only 44% and 12% when the Ptch allele was inactivated at E11.5 and E13.5, respectively. Induction of the Ptch mutation at E9.5 did also significantly shorten ERMS-free survival and increased tumor multiplicity compared with tumor induction at E11.5 and E13.5. Interestingly, we observed a more that 10-fold reduction of ERMS incidence when the Ptch mutation was specifically introduced in Myf5-expressing cells, which is the myogenic factor expressed in all muscle cells at E9.5. We conclude that Myf5-negative cells are more susceptible to ERMS development than Myf5-positive embryonic precursors. As the propensity to undergo tumorigenic transformation declined with age, concomitant with the increase of stably committed muscle cells, it seems likely that the Ptch mutation favors tumor formation in progenitor cells, which have not yet acquired a muscle cell fate.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  • Asakura A, Tapscott SJ . (1998). Apoptosis of epaxial myotome in Danforth's short-tail (Sd) mice in somites that form following notochord degeneration. Dev Biol 203: 276–289.

    Article  CAS  Google Scholar 

  • Biressi S, Molinaro M, Cossu G . (2007a). Cellular heterogeneity during vertebrate skeletal muscle development. Dev Biol 308: 281–293.

    Article  CAS  Google Scholar 

  • Biressi S, Tagliafico E, Lamorte G, Monteverde S, Tenedini E, Roncaglia E et al. (2007b). Intrinsic phenotypic diversity of embryonic and fetal myoblasts is revealed by genome-wide gene expression analysis on purified cells. Dev Biol 304: 633–651.

    Article  CAS  Google Scholar 

  • Braun T, Bober E, Rudnicki MA, Jaenisch R, Arnold HH . (1994). MyoD expression marks the onset of skeletal myogenesis in Myf-5 mutant mice. Development 120: 3083–3092.

    CAS  PubMed  Google Scholar 

  • Braun T, Rudnicki MA, Arnold HH, Jaenisch R . (1992). Targeted inactivation of the muscle regulatory gene Myf-5 results in abnormal rib development and perinatal death. Cell 71: 369–382.

    Article  CAS  Google Scholar 

  • Calzada-Wack J, Kappler R, Schnitzbauer U, Richter T, Nathrath M, Rosemann M et al. (2002). Unbalanced overexpression of the mutant allele in murine Patched mutants. Carcinogenesis 23: 727–734.

    Article  CAS  Google Scholar 

  • Danielian PS, Muccino D, Rowitch DH, Michael SK, McMahon AP . (1998). Modification of gene activity in mouse embryos in utero by a tamoxifen-inducible form of Cre recombinase. Curr Biol 8: 1323–1326.

    Article  CAS  Google Scholar 

  • Diwan BA, Anderson LM, Ward JM . (1997). Proliferative lesions of oviduct and uterus in CD-1 mice exposed prenatally to tamoxifen. Carcinogenesis 18: 2009–2014.

    Article  CAS  Google Scholar 

  • Ecke I, Petry F, Rosenberger A, Tauber S, Monkemeyer S, Hess I et al. (2009). Antitumor effects of a combined 5-aza-2′deoxycytidine and valproic acid treatment on rhabdomyosarcoma and medulloblastoma in Ptch mutant mice. Cancer Res 69: 887–895.

    Article  CAS  Google Scholar 

  • Evangelista M, Tian H, de Sauvage FJ . (2006). The hedgehog signaling pathway in cancer. Clin Cancer Res 12: 5924–5928.

    Article  CAS  Google Scholar 

  • Gensch N, Borchardt T, Schneider A, Riethmacher D, Braun T . (2008). Different autonomous myogenic cell populations revealed by ablation of Myf5-expressing cells during mouse embryogenesis. Development 135: 1597–1604.

    Article  CAS  Google Scholar 

  • Goodrich LV, Johnson RL, Milenkovic L, McMahon JA, Scott MP . (1996). Conservation of the hedgehog/patched signaling pathway from flies to mice: induction of a mouse patched gene by Hedgehog. Genes Dev 10: 301–312.

    Article  CAS  Google Scholar 

  • Gorlin RJ . (1987). Nevoid basal-cell carcinoma syndrome. Medicine (Baltimore) 66: 98–113.

    Article  CAS  Google Scholar 

  • Gustafsson MK, Pan H, Pinney DF, Liu Y, Lewandowski A, Epstein DJ et al. (2002). Myf5 is a direct target of long-range Shh signaling and Gli regulation for muscle specification. Genes Dev 16: 114–126.

    Article  CAS  Google Scholar 

  • Hahn H, Christiansen J, Wicking C, Zaphiropoulos PG, Chidambaram A, Gerrard B et al. (1996). A mammalian patched homolog is expressed in target tissues of sonic hedgehog and maps to a region associated with developmental abnormalities. J Biol Chem 271: 12125–12128.

    Article  CAS  Google Scholar 

  • Hahn H, Nitzki F, Schorban T, Hemmerlein B, Threadgill D, Rosemann M . (2004). Genetic mapping of a Ptch1-associated rhabdomyosarcoma susceptibility locus on mouse chromosome 2. Genomics 84: 853–858.

    Article  CAS  Google Scholar 

  • Hahn H, Wojnowski L, Zimmer AM, Hall J, Miller G, Zimmer A . (1998). Rhabdomyosarcomas and radiation hypersensitivity in a mouse model of Gorlin syndrome. Nature Med 4: 619–622.

    Article  CAS  Google Scholar 

  • Hameyer D, Loonstra A, Eshkind L, Schmitt S, Antunes C, Groen A et al. (2007). Toxicity of ligand-dependant Cre-recombinases and generation of a conditional Cre-deleter mouse allowing mosaic recombination in peripheral tissues. Physiol Genomics 31: 32–41.

    Article  CAS  Google Scholar 

  • Hooper JE, Scott MP . (2005). Communicating with Hedgehogs. Nat Rev Mol Cell Biol 6: 306–317.

    Article  CAS  Google Scholar 

  • Kappler R, Bauer R, Calzada-Wack J, Rosemann M, Hemmerlein B, Hahn H . (2004). Profiling the molecular difference between Patched- and p53-dependent rhabdomyosarcoma. Oncogene 23: 8785–8795.

    Article  CAS  Google Scholar 

  • Keller C, Arenkiel BR, Coffin CM, El-Bardeesy N, DePinho RA, Capecchi MR . (2004a). Alveolar rhabdomyosarcomas in conditional Pax3: Fkhr mice: cooperativity of Ink4a/ARF and Trp53 loss of function. Genes Dev 18: 2614–2626.

    Article  CAS  Google Scholar 

  • Keller C, Hansen MS, Coffin CM, Capecchi MR . (2004b). Pax3: Fkhr interferes with embryonic Pax3 and Pax7 function: implications for alveolar rhabdomyosarcoma cell of origin. Genes Dev 18: 2608–2613.

    Article  CAS  Google Scholar 

  • Kruger M, Mennerich D, Fees S, Schafer R, Mundlos S, Braun T . (2001). Sonic hedgehog is a survival factor for hypaxial muscles during mouse development. Development 128: 743–752.

    CAS  PubMed  Google Scholar 

  • Langenau DM, Keefe MD, Storer NY, Guyon JR, Kutok JL, Le X et al. (2007). Effects of RAS on the genesis of embryonal rhabdomyosarcoma. Genes Dev 21: 1382–1395.

    Article  CAS  Google Scholar 

  • McDermott A, Gustafsson M, Elsam T, Hui CC, Emerson Jr CP, Borycki AG . (2005). Gli2 and Gli3 have redundant and context-dependent function in skeletal muscle formation. Development 132: 345–357.

    Article  CAS  Google Scholar 

  • Merlino G, Helman LJ . (1999). Rhabdomyosarcoma--working out the pathways. Oncogene 18: 5340–5348.

    Article  CAS  Google Scholar 

  • Merlino G, Khanna C . (2007). Fishing for the origins of cancer. Genes Dev 21: 1275–1279.

    Article  CAS  Google Scholar 

  • Nagao K, Togawa N, Fujii K, Uchikawa H, Kohno Y, Yamada M et al. (2005). Detecting tissue-specific alternative splicing and disease-associated aberrant splicing of the PTCH gene with exon junction microarrays. Hum Mol Genet 14: 3379–3388.

    Article  CAS  Google Scholar 

  • Resende TP, Ferreira M, Teillet MA, Tavares AT, Andrade RP, Palmeirim I . (2010). Sonic hedgehog in temporal control of somite formation. Proc Natl Acad Sci USA 107: 12907–12912.

    Article  CAS  Google Scholar 

  • Rivera OJ, Cho MJ, Arenkiel BR, Milenkovic L, Capecchi MR, Scott MP et al. (2006). Medulloblastoma but not rhabdomyosarcoma in a conditional mouse model of Patched1 haploinsufficiency. Abstract no. 2742; 97th AACR Annual Meeting; 1–5 April 2006; Washington, DC.

  • Tallquist MD, Weismann KE, Hellstrom M, Soriano P . (2000). Early myotome specification regulates PDGFA expression and axial skeleton development. Development 127: 5059–5070.

    CAS  PubMed  Google Scholar 

  • Taniguchi E, Cho MJ, Arenkiel BR, Hansen MS, Rivera OJ, McCleish AT et al. (2009). Bortezomib reverses a post-translational mechanism of tumorigenesis for patched1 haploinsufficiency in medulloblastoma. Pediatr Blood Cancer 53: 136–144.

    Article  Google Scholar 

  • Tostar U, Malm CJ, Meis-Kindblom JM, Kindblom LG, Toftgard R, Unden AB . (2006). Deregulation of the hedgehog signalling pathway: a possible role for the PTCH and SUFU genes in human rhabdomyoma and rhabdomyosarcoma development. J Pathol 208: 17–25.

    Article  CAS  Google Scholar 

  • Uhmann A, Dittmann K, Nitzki F, Dressel R, Koleva M, Frommhold A et al. (2007). The Hedgehog receptor Patched controls lymphoid lineage commitment. Blood 110: 1814–1823.

    Article  CAS  Google Scholar 

  • Uhmann A, Ferch U, Bauer R, Tauber S, Arziman Z, Chen C et al. (2005). A model for PTCH1/Ptch1-associated tumors comprising mutational inactivation and gene silencing. Int J Oncol 27: 1567–1575.

    CAS  PubMed  Google Scholar 

  • Varjosalo M, Taipale J . (2008). Hedgehog: functions and mechanisms. Genes Dev 22: 2454–2472.

    Article  CAS  Google Scholar 

  • Yang ZJ, Ellis T, Markant SL, Read TA, Kessler JD, Bourboulas M et al. (2008). Medulloblastoma can be initiated by deletion of Patched in lineage-restricted progenitors or stem cells. Cancer Cell 14: 135–145.

    Article  CAS  Google Scholar 

  • Zibat A, Missiaglia E, Rosenberger A, Pritchard-Jones K, Shipley J, Hahn H et al. (2010). Activation of the hedgehog pathway confers a poor prognosis in embryonal and fusion gene-negative alveolar rhabdomyosarcoma. Oncogene 29: 6323–6330.

    Article  CAS  Google Scholar 

  • Zibat A, Uhmann A, Nitzki F, Wijgerde M, Frommhold A, Heller T et al. (2009). Time-point and dosage of gene inactivation determine the tumor spectrum in conditional Ptch knockouts. Carcinogenesis 30: 918–926.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Susan Peter and Stefan Wolf for excellent animal care and Simone Fulda for critical comments on the manuscript. This work was supported by the grant 2003.112.3 of the Wilhelm Sander Stiftung to HH.

Author information

Authors and Affiliations

  1. Department of Human Genetics, Universitätsmedizin Göttingen, Göttingen, Germany

    F Nitzki, A Zibat, A Frommhold & H Hahn

  2. Department of Cardiac Development and Remodelling, Max-Planck-Institute for Heart- und Lung Research, Bad Nauheim, Germany

    A Schneider & T Braun

  3. Department of Neuropathology, Universitätsmedizin Göttingen, Göttingen, Germany

    W Schulz-Schaeffer

Authors
  1. F Nitzki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A Zibat
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Frommhold
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Schneider
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. W Schulz-Schaeffer
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. T Braun
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H Hahn
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to H Hahn.

Ethics declarations

Competing interests

The authors do not have a conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nitzki, F., Zibat, A., Frommhold, A. et al. Uncommitted precursor cells might contribute to increased incidence of embryonal rhabdomyosarcoma in heterozygous Patched1-mutant mice. Oncogene 30, 4428–4436 (2011). https://doi.org/10.1038/onc.2011.157

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2011.157

Keywords

This article is cited by

Search

Advanced search

Quick links