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GOLPH3 modulates mTOR signalling and rapamycin sensitivity in cancer

Nature volume 459, pages 1085–1090 (2009)Cite this article

Abstract

Genome-wide copy number analyses of human cancers identified a frequent 5p13 amplification in several solid tumour types, including lung (56%), ovarian (38%), breast (32%), prostate (37%) and melanoma (32%). Here, using integrative analysis of a genomic profile of the region, we identify a Golgi protein, GOLPH3, as a candidate targeted for amplification. Gain- and loss-of-function studies in vitro and in vivo validated GOLPH3 as a potent oncogene. Physically, GOLPH3 localizes to the trans-Golgi network and interacts with components of the retromer complex, which in yeast has been linked to target of rapamycin (TOR) signalling. Mechanistically, GOLPH3 regulates cell size, enhances growth-factor-induced mTOR (also known as FRAP1) signalling in human cancer cells, and alters the response to an mTOR inhibitor in vivo. Thus, genomic and genetic, biological, functional and biochemical data in yeast and humans establishes GOLPH3 as a new oncogene that is commonly targeted for amplification in human cancer, and is capable of modulating the response to rapamycin, a cancer drug in clinical use.

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Figure 1: Genomic characterization of 5p13 amplification.
Figure 2: Functional validation of GOLPH3.
Figure 3: GOLPH3 interacts with VPS35 and correlates with S6K phosphorylation.
Figure 4: GOLPH3 modulates cell size and phosphorylation status of mTOR substrates.
Figure 5: The in vivo GOLPH3 growth advantage is abrogated by treatment with rapamycin.

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References

  1. Manning, B. D. & Cantley, L. C. AKT/PKB signaling: navigating downstream. Cell 129, 1261–1274 (2007)

    Article  CAS  Google Scholar 

  2. Wullschleger, S., Loewith, R. & Hall, M. N. TOR signaling in growth and metabolism. Cell 124, 471–484 (2006)

    Article  CAS  Google Scholar 

  3. Shima, H. et al. Disruption of the p70s6k/p85s6k gene reveals a small mouse phenotype and a new functional S6 kinase. EMBO J. 17, 6649–6659 (1998)

    Article  CAS  Google Scholar 

  4. Montagne, J. et al. Drosophila S6 kinase: a regulator of cell size. Science 285, 2126–2129 (1999)

    Article  CAS  Google Scholar 

  5. Oldham, S., Montagne, J., Radimerski, T., Thomas, G. & Hafen, E. Genetic and biochemical characterization of dTOR, the Drosophila homolog of the target of rapamycin. Genes Dev. 14, 2689–2694 (2000)

    Article  CAS  Google Scholar 

  6. Zhang, H., Stallock, J. P., Ng, J. C., Reinhard, C. & Neufeld, T. P. Regulation of cellular growth by the Drosophila target of rapamycin dTOR. Genes Dev. 14, 2712–2724 (2000)

    Article  CAS  Google Scholar 

  7. Fingar, D. C. & Blenis, J. Target of rapamycin (TOR): an integrator of nutrient and growth factor signals and coordinator of cell growth and cell cycle progression. Oncogene 23, 3151–3171 (2004)

    Article  CAS  Google Scholar 

  8. Guertin, D. A. & Sabatini, D. M. Defining the role of mTOR in cancer. Cancer Cell 12, 9–22 (2007)

    Article  CAS  Google Scholar 

  9. Yang, Q. & Guan, K. L. Expanding mTOR signaling. Cell Res. 17, 666–681 (2007)

    Article  CAS  Google Scholar 

  10. Abraham, R. T. & Wiederrecht, G. J. Immunopharmacology of rapamycin. Annu. Rev. Immunol. 14, 483–510 (1996)

    Article  CAS  Google Scholar 

  11. Sabatini, D. M. mTOR and cancer: insights into a complex relationship. Nature Rev. Cancer 6, 729–734 (2006)

    Article  CAS  Google Scholar 

  12. Garraway, L. A. et al. Integrative genomic analyses identify MITF as a lineage survival oncogene amplified in malignant melanoma. Nature 436, 117–122 (2005)

    Article  ADS  CAS  Google Scholar 

  13. Wu, C. C. et al. GMx33: a novel family of trans-Golgi proteins identified by proteomics. Traffic 1, 963–975 (2000)

    CAS  PubMed  Google Scholar 

  14. Bell, A. W. et al. Proteomics characterization of abundant Golgi membrane proteins. J. Biol. Chem. 276, 5152–5165 (2001)

    Article  CAS  Google Scholar 

  15. Snyder, C. M., Mardones, G. A., Ladinsky, M. S. & Howell, K. E. GMx33 associates with the trans-Golgi matrix in a dynamic manner and sorts within tubules exiting the Golgi. Mol. Biol. Cell 17, 511–524 (2006)

    Article  CAS  Google Scholar 

  16. Bonifacino, J. S. & Hurley, J. H. Retromer. Curr. Opin. Cell Biol. 20, 427–436 (2008)

    Article  CAS  Google Scholar 

  17. Xie, M. W. et al. Insights into TOR function and rapamycin response: chemical genomic profiling by using a high-density cell array method. Proc. Natl Acad. Sci. USA 102, 7215–7220 (2005)

    Article  ADS  CAS  Google Scholar 

  18. Camp, R. L., Chung, G. G. & Rimm, D. L. Automated subcellular localization and quantification of protein expression in tissue microarrays. Nature Med. 8, 1323–1327 (2002)

    Article  CAS  Google Scholar 

  19. Fingar, D. C., Salama, S., Tsou, C., Harlow, E. & Blenis, J. Mammalian cell size is controlled by mTOR and its downstream targets S6K1 and 4EBP1/eIF4E. Genes Dev. 16, 1472–1487 (2002)

    Article  CAS  Google Scholar 

  20. Burnett, P. E., Barrow, R. K., Cohen, N. A., Snyder, S. H. & Sabatini, D. M. RAFT1 phosphorylation of the translational regulators p70 S6 kinase and 4E-BP1. Proc. Natl Acad. Sci. USA 95, 1432–1437 (1998)

    Article  ADS  CAS  Google Scholar 

  21. Isotani, S. et al. Immunopurified mammalian target of rapamycin phosphorylates and activates p70 S6 kinase α in vitro . J. Biol. Chem. 274, 34493–34498 (1999)

    Article  CAS  Google Scholar 

  22. Hresko, R. C. & Mueckler, M. mTOR·RICTOR is the Ser473 kinase for Akt/protein kinase B in 3T3–L1 adipocytes. J. Biol. Chem. 280, 40406–40416 (2005)

    Article  CAS  Google Scholar 

  23. Sarbassov, D. D., Guertin, D. A., Ali, S. M. & Sabatini, D. M. Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307, 1098–1101 (2005)

    Article  ADS  CAS  Google Scholar 

  24. Chiu, V. K. et al. Ras signalling on the endoplasmic reticulum and the Golgi. Nature Cell Biol. 4, 343–350 (2002)

    Article  CAS  Google Scholar 

  25. Eaton, S. Retromer retrieves wntless. Dev. Cell 14, 4–6 (2008)

    Article  CAS  Google Scholar 

  26. Korolchuk, V. I. et al. Drosophila Vps35 function is necessary for normal endocytic trafficking and actin cytoskeleton organisation. J. Cell Sci. 120, 4367–4376 (2007)

    Article  CAS  Google Scholar 

  27. Schmitz, K. R. et al. Golgi localization of glycosyltransferases requires a Vps74p oligomer. Dev. Cell 14, 523–534 (2008)

    Article  CAS  Google Scholar 

  28. Tu, L., Tai, W. C., Chen, L. & Banfield, D. K. Signal-mediated dynamic retention of glycosyltransferases in the Golgi. Science 321, 404–407 (2008)

    Article  ADS  CAS  Google Scholar 

  29. Ohtsubo, K. & Marth, J. D. Glycosylation in cellular mechanisms of health and disease. Cell 126, 855–867 (2006)

    Article  CAS  Google Scholar 

  30. Takahashi, M., Tsuda, T., Ikeda, Y., Honke, K. & Taniguchi, N. Role of N-glycans in growth factor signaling. Glycoconj. J. 20, 207–212 (2004)

    Article  CAS  Google Scholar 

  31. Maser, R. S. et al. Chromosomally unstable mouse tumours have genomic alterations similar to diverse human cancers. Nature 447, 966–971 (2007)

    Article  ADS  CAS  Google Scholar 

  32. Satyamoorthy, K. et al. Melanoma cell lines from different stages of progression and their biological and molecular analyses. Melanoma Res. 7 (suppl. 2). S35–S42 (1997)

    Article  CAS  Google Scholar 

  33. Tonon, G. et al. High-resolution genomic profiles of human lung cancer. Proc. Natl Acad. Sci. USA 102, 9625–9630 (2005)

    Article  ADS  CAS  Google Scholar 

Download references

Acknowledgements

We thank R. DePinho for critical reading of the manuscript, and L. Cantley, as well as members of the Chin laboratory, for helpful discussion. We thank H. Ying for assistance with confocal microscopy. The rabbit polyclonal antibody against human GOLPH3 was kindly provided by J. J. Bergeron of McGill University. Mouse monoclonal antibody against human GOLPH3, C19, was generated at the Dana-Farber/Harvard Cancer Center Monoclonal Antibody Core Facility. K.L.S. is at present supported by a Postdoctoral Fellowship from the American Cancer Society (PF-07-039-01-CSM), and K.L.S. and O.K. were previously supported by a National Institutes of Health (NIH) Training Grant appointment in the Department of Dermatology at Brigham and Women’s Hospital (5-T32-AR07098-31). K.-K.W. was supported by a Program of Research Excellence (SPORE) grant (P50 CA090578) and NIH grants (R01 AG2400401; R01 CA122794). The AQUA immunofluorescence study was supported by a grant from the NIH to D.L.R. (RO-1 CA 114277). This work is primarily supported by grants from the NIH to L.C. (RO1 CA93947; P50 CA93683).

Author Contribution O.K. identified GOLPH3 as an oncogene target of 5p13; K.L.S. performed oncogene validation and mechanistic studies; E.I. and A.P. performed TMA FISH analysis; H.R.W. and D.E.F. performed the HMEL anchorage-independent growth assay; S.D. and J.W. assisted with immunofluorescence assays and provided technical support; M.W. performed the co-immunoprecipitation assays; V.A. and D.L.R. performed AQUA analysis; S.C. assisted with cell culture studies; A.S. aided phospholipid and biochemical analyses; M.-C.L. and K.-K.W. performed rapamycin xenograft assays; Y.X. performed computational analyses; T.F. and J.H. provided technical support; L.C. supervised the experiments and data interpretation; K.L.S. and L.C. wrote the manuscript.

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Author notes

  1. Kenneth L. Scott and Omar Kabbarah: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Oncology,,

    Kenneth L. Scott, Omar Kabbarah, Mei-Chih Liang, Joyce Wu, Sabin Dhakal, Min Wu, Shujuan Chen, Tamar Feinberg, Joseph Huang, Kwok-Kin Wong & Lynda Chin

  2. Belfer Institute for Applied Cancer Science,,

    Elena Ivanova, Yonghong Xiao, Alexei Protopopov & Lynda Chin

  3. Department of Pediatric Oncology, Dana-Farber Cancer Institute,

    Hans R. Widlund & David E. Fisher

  4. Ludwig Center at Dana-Farber/Harvard Cancer Center, Boston, Massachusetts 02115, USA ,

    Mei-Chih Liang & Kwok-Kin Wong

  5. Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06520, USA,

    Valsamo Anagnostou & David L. Rimm

  6. Department of Systems Biology, Harvard Medical School,

    Abdel Saci

  7. Department of Dermatology, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA,

    Hans R. Widlund & Lynda Chin

  8. Department of Dermatology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA,

    David E. Fisher

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Correspondence to Lynda Chin.

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Scott, K., Kabbarah, O., Liang, MC. et al. GOLPH3 modulates mTOR signalling and rapamycin sensitivity in cancer. Nature 459, 1085–1090 (2009). https://doi.org/10.1038/nature08109

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