Article | Published:

Androgen-induced TOP2B-mediated double-strand breaks and prostate cancer gene rearrangements

Nature Genetics volume 42, pages 668675 (2010) | Download Citation

This article has been updated

Abstract

DNA double-strand breaks (DSBs) can lead to the development of genomic rearrangements, which are hallmarks of cancer. Fusions between TMPRSS2, encoding the transmembrane serine protease isoform 2, and ERG, encoding the v-ets erythroblastosis virus E26 oncogene homolog, are among the most common oncogenic rearrangements observed in human cancer. We show that androgen signaling promotes co-recruitment of androgen receptor and topoisomerase II beta (TOP2B) to sites of TMPRSS2-ERG genomic breakpoints, triggering recombinogenic TOP2B-mediated DSBs. Furthermore, androgen stimulation resulted in de novo production of TMPRSS2-ERG fusion transcripts in a process that required TOP2B and components of the DSB repair machinery. Finally, unlike normal prostate epithelium, prostatic intraepithelial neoplasia cells showed strong coexpression of androgen receptor and TOP2B. These findings implicate androgen-induced TOP2B-mediated DSBs in generating TMPRSS2-ERG rearrangements.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Change history

  • 18 July 2010

    In the version of this article initially published online, there were four sentences (in the Results on pages 2 and 3, in the legend to Figure 3 and in the Online Methods) containing minor errors. These errors have been corrected for the print, PDF and HTML versions of this article.

Accessions

Gene Expression Omnibus

References

  1. 1.

    Chromosomal translocations: revisited yet again. Blood 112, 2183–2189 (2008).

  2. 2.

    , & The impact of translocations and gene fusions on cancer causation. Nat. Rev. Cancer 7, 233–245 (2007).

  3. 3.

    et al. DNA double-strand breaks, chromosomal rearrangements, and genomic instability. Mutat. Res. 404, 125–128 (1998).

  4. 4.

    & Frequent chromosomal translocations induced by DNA double-strand breaks. Nature 405, 697–700 (2000).

  5. 5.

    , & Recurrent gene fusions in prostate cancer. Nat. Rev. Cancer 8, 497–511 (2008).

  6. 6.

    et al. Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science 310, 644–648 (2005).

  7. 7.

    et al. Cancer statistics, 2009. CA Cancer J. Clin. 59, 225–249 (2009).

  8. 8.

    , & Balanced chromosome aberrations in leukemias following chemotherapy with DNA-topoisomerase II inhibitors. J. Clin. Oncol. 16, 1897–1898 (1998).

  9. 9.

    , & The secondary leukemias: challenges and research directions. J. Natl. Cancer Inst. 88, 407–418 (1996).

  10. 10.

    & Chromatin structural elements and chromosomal translocations in leukemia. DNA Repair (Amst.) 5, 1282–1297 (2006).

  11. 11.

    , , , & Dietary bioflavonoids induce cleavage in the MLL gene and may contribute to infant leukemia. Proc. Natl. Acad. Sci. USA 97, 4790–4795 (2000).

  12. 12.

    et al. A topoisomerase IIβ-mediated dsDNA break required for regulated transcription. Science 312, 1798–1802 (2006).

  13. 13.

    et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. USA 102, 15545–15550 (2005).

  14. 14.

    , , & Gene Set Enrichment Analysis Made Simple (The Berkeley Electronic Press, 2009).

  15. 15.

    , & Newly replicated DNA is associated with DNA topoisomerase II in cultured rat prostatic adenocarcinoma cells. Nature 322, 187–189 (1986).

  16. 16.

    et al. Copy number analysis indicates monoclonal origin of lethal metastatic prostate cancer. Nat. Med. 15, 559–565 (2009).

  17. 17.

    et al. Distinct genomic aberrations associated with ERG rearranged prostate cancer. Genes Chromosom. Cancer 48, 366–380 (2009).

  18. 18.

    et al. Multiple genomic alterations on 21q22 predict various TMPRSS2/ERG fusion transcripts in human prostate cancers. Genes Chromosom. Cancer 46, 972–980 (2007).

  19. 19.

    et al. A hierarchical network of transcription factors governs androgen receptor-dependent prostate cancer growth. Mol. Cell 27, 380–392 (2007).

  20. 20.

    , , & Capturing chromosome conformation. Science 295, 1306–1311 (2002).

  21. 21.

    , , & Gamma-H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic Acids Res. 36, 5678–5694 (2008).

  22. 22.

    , & Roles of ATM and NBS1 in chromatin structure modulation and DNA double-strand break repair. Nat. Cell Biol. 9, 683–690 (2007).

  23. 23.

    Ataxia-telangiectasia: from a rare disorder to a paradigm for cell signalling and cancer. Nat. Rev. Mol. Cell Biol. 9, 759–769 (2008).

  24. 24.

    et al. Complex patterns of ETS gene alteration arise during cancer development in the human prostate. Oncogene 27, 1993–2003 (2008).

  25. 25.

    et al. TMPRSS2-ERG fusion prostate cancer: an early molecular event associated with invasion. Am. J. Surg. Pathol. 31, 882–888 (2007).

  26. 26.

    et al. Nuclear receptor-induced chromosomal proximity and DNA breaks underlie specific translocations in cancer. Cell 139, 1069–1083 (2009).

  27. 27.

    , , & Prooxidant-antioxidant shift induced by androgen treatment of human prostate carcinoma cells. J. Natl. Cancer Inst. 89, 40–48 (1997).

  28. 28.

    et al. Inflammation in prostate carcinogenesis. Nat. Rev. Cancer 7, 256–269 (2007).

  29. 29.

    et al. Activation of topoisomerase II-mediated excision of chromosomal DNA loops during oxidative stress. Genes Dev. 13, 1553–1560 (1999).

  30. 30.

    et al. Roles of DNA topoisomerase II isozymes in chemotherapy and secondary malignancies. Proc. Natl. Acad. Sci. USA 104, 11014–11019 (2007).

  31. 31.

    , & A model for tumor cell killing by topoisomerase poisons. Cancer Cells 2, 23–27 (1990).

  32. 32.

    , & Prostate cancer. N. Engl. J. Med. 349, 366–381 (2003).

  33. 33.

    , & Role of programmed (apoptotic) cell death during the progression and therapy for prostate cancer. Prostate 28, 251–265 (1996).

  34. 34.

    Does prostate cancer co-opt the developmental program? Differentiation 76, 736–744 (2008).

  35. 35.

    , & Is the Achilles' heel for prostate cancer therapy a gain of function in androgen receptor signaling? J. Clin. Endocrinol. Metab. 88, 2972–2982 (2003).

  36. 36.

    & Genetic changes and DNA damage responses in the prostate. Prostate 68, 902–918 (2008).

  37. 37.

    et al. Aberrant ERG expression cooperates with loss of PTEN to promote cancer progression in the prostate. Nat. Genet. 41, 619–624 (2009).

  38. 38.

    et al. Cooperativity of TMPRSS2-ERG with PI3-kinase pathway activation in prostate oncogenesis. Nat. Genet. 41, 524–526 (2009).

  39. 39.

    et al. A causal role for ERG in neoplastic transformation of prostate epithelium. Proc. Natl. Acad. Sci. USA 105, 2105–2110 (2008).

  40. 40.

    et al. Two unique novel prostate-specific and androgen-regulated fusion partners of ETV4 in prostate cancer. Cancer Res. 68, 3094–3098 (2008).

  41. 41.

    et al. Induced chromosomal proximity and gene fusions in prostate cancer. Science 326, 1230 (2009).

  42. 42.

    & Genome instability: a mechanistic view of its causes and consequences. Nat. Rev. Genet. 9, 204–217 (2008).

  43. 43.

    et al. High-mobility group chromatin proteins 1 and 2 functionally interact with steroid hormone receptors to enhance their DNA binding in vitro and transcriptional activity in mammalian cells. Mol. Cell. Biol. 18, 4471–4487 (1998).

  44. 44.

    , & Spatial and temporal recruitment of androgen receptor and its coactivators involves chromosomal looping and polymerase tracking. Mol. Cell 19, 631–642 (2005).

  45. 45.

    , & 3C technology: analyzing the spatial organization of genomic loci in vivo. Methods Enzymol. 375, 493–507 (2004).

  46. 46.

    , & Eukaryotic topoisomerase II preferentially cleaves alternating purine-pyrimidine repeats. Nucleic Acids Res. 18, 1–11 (1990).

  47. 47.

    et al. Human papillomavirus E6 and E7 oncoproteins alter cell cycle progression but not radiosensitivity of carcinoma cells treated with low-dose-rate radiation. Int. J. Radiat. Oncol. Biol. Phys. 37, 145–154 (1997).

  48. 48.

    et al. Nuclear MYC protein overexpression is an early alteration in human prostate carcinogenesis. Mod. Pathol. 21, 1156–1167 (2008).

  49. 49.

    , , & A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19, 185–193 (2003).

  50. 50.

    et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80 (2004).

  51. 51.

    et al. Interaction and functional interference of glucocorticoid receptor and SOCS1. J. Biol. Chem. 283, 22089–22096 (2008).

  52. 52.

    et al. Association of guanine nucleotide-exchange protein BIG1 in HepG2 cell nuclei with nucleolin, U3 snoRNA, and fibrillarin. Proc. Natl. Acad. Sci. USA 105, 3357–3361 (2008).

Download references

Acknowledgements

We thank D. Coffey for helpful comments and C. Heaphy, H. Zhang and L. Dasko-Vincent from the SKCCC Cell Imaging Core Facility for technical support. We also thank the Brady Urological Research Institute Prostate Specimen Repository for providing TMA sections. This work was supported by funding from the NIH/NCI, Department of Defense PCRP, Prostate Cancer Foundation, Maryland Cigarette Restitution Fund and the Patrick C. Walsh Prostate Cancer Research Fund/Dr. and Mrs. Peter S. Bing Scholarship.

Author information

Author notes

    • Martin J Aryee
    •  & Antoun Toubaji

    These authors contributed equally to this work.

Affiliations

  1. Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, Maryland, USA.

    • Michael C Haffner
    • , Martin J Aryee
    • , David M Esopi
    • , William B Isaacs
    • , Alan K Meeker
    • , George Netto
    • , Angelo M De Marzo
    • , William G Nelson
    •  & Srinivasan Yegnasubramanian
  2. Department of Biostatistics, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland, USA.

    • Martin J Aryee
  3. Department of Pathology, Johns Hopkins University, Baltimore, Maryland, USA.

    • Antoun Toubaji
    • , Roula Albadine
    • , Bora Gurel
    • , G Steven Bova
    • , Alan K Meeker
    • , George Netto
    • , Angelo M De Marzo
    •  & William G Nelson
  4. Brady Urological Institute, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.

    • William B Isaacs
    • , Alan K Meeker
    • , George Netto
    • , Angelo M De Marzo
    •  & William G Nelson
  5. Center for Cancer Genomics, Wake Forest University Health Sciences, Winston-Salem, North Carolina, USA.

    • Wennuan Liu
    •  & Jianfeng Xu

Authors

  1. Search for Michael C Haffner in:

  2. Search for Martin J Aryee in:

  3. Search for Antoun Toubaji in:

  4. Search for David M Esopi in:

  5. Search for Roula Albadine in:

  6. Search for Bora Gurel in:

  7. Search for William B Isaacs in:

  8. Search for G Steven Bova in:

  9. Search for Wennuan Liu in:

  10. Search for Jianfeng Xu in:

  11. Search for Alan K Meeker in:

  12. Search for George Netto in:

  13. Search for Angelo M De Marzo in:

  14. Search for William G Nelson in:

  15. Search for Srinivasan Yegnasubramanian in:

Contributions

M.C.H. executed and analyzed all experiments and assisted in writing the manuscript. M.J.A. analyzed microarray data and assisted with statistical analysis of data. A.T., R.A., B.G., A.K.M., G.N. and A.M.D.M. assisted with execution and analysis of FISH, immunostaining and pathology experiments. D.M.E. assisted with execution of experiments. W.B.I., G.S.B., W.L. and J.X. contributed to analysis of microarray data in determination of prostate cancer TMPRSS2-ERG genomic breakpoints. W.G.N. assisted in experimental design and analysis and contributed to writing the manuscript. S.Y. conceived the study together with W.G.N., assisted in experimental design, execution and analysis and wrote the manuscript. All authors assisted in editing the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to William G Nelson or Srinivasan Yegnasubramanian.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–19 and Supplementary Tables 1–4.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ng.613

Further reading