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Androgen receptor activation results in metabolite signatures of an aggressive prostate cancer phenotype: an NMR-based metabonomics study

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Abstract

Metabolomic studies have proven to provide a unique perspective of the cellular dysfunction developing as a result of prostate cancer (PCa) onset and progression, facilitated primarily by mass spectrometry (MS) and nuclear magnetic resonance (NMR) techniques. PCa develops as an androgen-dependent disease with the expression of the androgen receptor (AR), where patient treatment typically involves androgen ablation therapy. In response, it is theorized that PCa transforms to an androgen-hypersensitive or androgen-independent state, where treatment options are severely reduced. Under the hypothesis that AR stimulation increases the aggressivity of pre-existing PCa, NMR spectroscopy was utilized in the delineation of the metabonomic response of an androgen-dependent PCa cell line (LnCAP) as a result of AR activation. Metabolite profiles were determined after 12, 24, and 48 h of exposure to methyltrienolone (R1881), an AR agonist. Principal components analysis revealed the relative myo-inositol and phosphocholine levels were severely altered after androgen treatment. Furthermore, univariate analysis revealed multiple metabolic perturbations in response to R1881 exposure, including amino acid, choline, the phosphocholine/glycerophosphocholine ratio, and UDP-coupled sugar metabolism, which are consistent with reported measurements between normal and PCa samples. These results suggest that androgen-sensitive PCa may transform to an aggressive phenotype upon AR activation.

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References

  • Ackerstaff, E., Pfug, B. R., Nelson, J. B., & Bhujwalla, Z. M. (2001). Detection of increased choline compounds with proton nuclear magnetic resonance spectroscopy subsequent to malignant transformation of human prostatic epithelial cells. Cancer Research, 61, 3599–3603.

    PubMed  CAS  Google Scholar 

  • Albers, M. J., Butler, T. N., Rahwa, I., Bao, N., Keshari, K. R., Swanson, M. G., et al. (2009). Evaluation of the ERETIC method as an improved quantitative reference for 1H HR-MAS spectroscopy of prostate tissue. Magnetic Resonance in Medicine, 61, 525–532.

    Article  PubMed  CAS  Google Scholar 

  • Baek, S. H., Ohgi, K. A., Nelson, C. A., Welsbie, D., Chen, C., Sawyers, C. L., et al. (2006). Ligand-specific allosteric regulation of coactivator functions of androgen receptor in prostate cancer cells. Proceedings of the National Academy of Sciences of the USA, 103, 3100–3105.

    Article  PubMed  CAS  Google Scholar 

  • Bao, B. Y., Chuang, B. F., Wang, Q., Sartor, O., Balk, S. P., Brown, M., et al. (2008). Androgen receptor mediates the expression of UDP-glucuronosyltransferase 2 BI5 and BI7 genes. The Prostate, 68, 839–848.

    Article  PubMed  CAS  Google Scholar 

  • Beckonert, O., Monnerjahn, J., Bonk, U., & Leibfritz, D. (2003). Visualizing metabolic changes in breast-cancer tissue using 1H-NMR spectroscopy and self-organizing maps. NMR in Biomedicine, 16, 1–11.

    Article  PubMed  CAS  Google Scholar 

  • Butcher, N. J., Tetlow, N. L., Cheung, C., Broadhurst, G. M., & Minchin, R. F. (2007). Induction of human arylamine N-acetyltransferase type i by androgens in human prostate cancer cells. Cancer Research, 67, 85–92.

    Article  PubMed  CAS  Google Scholar 

  • Cai, C., Wang, H., Xu, Y., Chen, S., & Balk, S. P. (2009). Reactivation of androgen receptor-regulated TMPRSS2:ERG gene expression in castration-resistant prostate cancer. Cancer Research, 69, 6027–6032.

    Article  PubMed  CAS  Google Scholar 

  • Cai, J., Kandagatla, P., Singareddy, R., Kropinski, A., Sheng, S., Cher, M. L., et al. (2010). Androgens induce functional CXCR4 through ERG factor expression in TMRPRSS2-ERG fusion-positive prostate cancer cells. Translational Oncology, 3, 195–203.

    PubMed  Google Scholar 

  • Chen, M., Tanner, M., Levine, A. C., Levina, E., Ohouo, P., & Buttyan, R. (2009). Androgenic regulation of hedgehog signaling pathway components in prostate cancer cells. Cell Cycle, 8, 149–157.

    Article  PubMed  CAS  Google Scholar 

  • Cheng, L. L., Wu, C., Smith, M. R., & Gonzalez, R. G. (2001). Non-destructive quantitation of spermine in human prostate tissue samples using HRMAS 1H NMR spectroscopy at 9.4 T. FEBS Letters, 494(1–2), 112–116.

    Article  PubMed  CAS  Google Scholar 

  • Cheng, L. L., Burns, M. A., Taylor, J. L., He, W., Halperin, E. F., McDougal, W. S., et al. (2005). Metabolic characterization of human prostate cancer with tissue magnetic resonance spectroscopy. Cancer Research, 65, 3030–3034.

    PubMed  CAS  Google Scholar 

  • Chipuk, J. E., Cornelius, S. C., Pultz, N. J., Jorgensen, J. S., Bonham, M. J., Kim, S. J., et al. (2002). The androgen receptor represses transforming growth factor-β signaling through interaction with Smad3. The Journal of Biological Chemistry, 277, 1240–1248.

    Article  PubMed  CAS  Google Scholar 

  • Church, D. R., Lee, E., Thompson, T. A., Basu, H. S., Ripple, M. O., Ariazi, R. A., et al. (2005). Induction of AP-I activity by androgen activation of the androgen receptor in LNCaP human prostate carcinoma cells. The Prostate, 63, 155–168.

    Article  PubMed  CAS  Google Scholar 

  • Costello, L. C., Franklin, R. B., & Narayan, P. (1999). Citrate in the diagnosis of prostate cancer. The Prostate, 38, 237–245.

    Article  PubMed  CAS  Google Scholar 

  • Damber, J. E., & Aus, G. (2008). Prostate cancer. Lancet, 37, 1710–1721.

    Article  Google Scholar 

  • Dieterle, F., Ross, A., Schlotterbeck, G., & Senn, H. (2006). Probabilistic quotient normalization as robust method to account for dilution of complex biological mixtures. Application in 1H NMR metabonomics. Analytical Chemistry, 78, 4281–4290.

    Article  PubMed  CAS  Google Scholar 

  • Feldman, B. J., & Feldman, D. (2001). The development of androgen-independent prostate cancer. Nature Reviews Cancer, 1, 34–45.

    Article  PubMed  CAS  Google Scholar 

  • Flammand, V., Zhao, H., & Peehl, D. M. (2010). Targeting monoamine oxidase A in advanced prostate cancer. Journal of Cancer Research and Clinical Oncology, 136, 1761–1771.

    Article  Google Scholar 

  • Gannon, P. O., Godin-Ethier, J., Hassler, M., Delvoye, N., Aversa, M., Poisson, A. O., et al. (2010). Androgen-regulated expression of arginase 1, arginase 2 and interleukin-8 in human prostate cancer. PLoS ONE, 5, doi:10.1371/journal.pone.0012107.

  • Gavrielides, M. V., Gonzalez-Guerrico, A. M., Riobo, N. A., & Kazanietz, M. G. (2006). Androgens regulate protein kinase Cδ transcription and modulate its apoptotic function in prostate cancer cells. Cancer Research, 66, 11792–11801.

    Article  PubMed  CAS  Google Scholar 

  • Hobisch, A., Eder, I. E., Putz, T., Horninger, W., Bartsch, G., Klocker, H., et al. (1998). Interleukin-6 regulates prostate-specific protein expression in prostate carcinoma cells by activation of the androgen receptor. Cancer Research, 58, 4640–4645.

    PubMed  CAS  Google Scholar 

  • Jackson, J. E. (1991). A user’s guide to principal components. New York: Wiley.

    Book  Google Scholar 

  • Jennbacken, K., Gustavsson, H., Tešan, T., Horn, M., Vallbo, C., Welén, K., et al. (2009). The prostatic environment suppresses growth of androgen-independent prostate cancer xenografts: An effect influenced by testosterone. The Prostate, 69, 1164–1175.

    Article  PubMed  CAS  Google Scholar 

  • Kimura, K., Markowski, M., Bowen, C., & Gelmann, E. P. (2001). Androgen blocks apoptosis of hormone-dependent prostate cancer cells. Cancer Research, 61, 5611–5618.

    PubMed  CAS  Google Scholar 

  • Lee, Y. C., Cheng, C. J., Huang, M., Bilen, M. A., Ye, X., Navone, N. M., et al. (2010). Androgen depletion up-regulates cadherin-11 expression in prostate cancer. Journal of Pathology, 221, 68–76.

    Article  PubMed  CAS  Google Scholar 

  • Levin, Y. S., Albers, M. J., Butler, T. N., Spielman, D., Peehl, D. M., & Kurhanewicz, J. (2009). Methods for metabolic evaluation of prostate cancer cells using proton and 13C HR-MAS spectroscopy and [3-13C] pyruvate as a metabolic substrate. Magnetic Resonance in Medicine, 62, 1091–1098.

    Article  PubMed  CAS  Google Scholar 

  • Loberg, R. D., McGregor, N., Ying, C., Sargent, E., & Pienta, K. J. (2007). In vivo evaluation of AT-101 (R-(-)-gossypol acetic acid) in androgen-independent growth of VCaP prostate cancer cells in combination with surgical castration. Neoplasia, 9, 1030–1037.

    Article  PubMed  CAS  Google Scholar 

  • Makkonen, H., Kauhanen, M., Jääskeläinen, T., & Palvimo, J. J. (2011). Androgen receptor amplification is reflected in the transcriptional responses of vertebral-cancer of the prostate cells. Molecular and Cellular Endocrinology, 331, 57–65.

    Article  PubMed  CAS  Google Scholar 

  • Mani, R. S., Tomlins, S. A., Callahan, K., Ghosh, A., Nyati, M. K., Varambally, S., et al. (2009). Induced chromosomal proximity and gene fusions in prostate cancer. Science, 326, 1230.

    Article  PubMed  CAS  Google Scholar 

  • Massie, C. E., Lynch, A., Ramos-Montoya, A., Boren, J., Stark, R., Fazli, L., et al. (2011). The androgen receptor fuels prostate cancer by regulating central metabolism and biosynthesis. The EMBO Journal, 1–15, doi:10.1038/emboj.2011.158.

  • Migliaccio, A., Castoria, G., Di Domenico, M., de Falco, A., Bilancio, A., Lombardi, M., et al. (2000). Steroid-induced androgen receptor-oestradiol receptor β-Src complex triggers prostate cancer cell proliferation. The EMBO Journal, 19, 5406–5417.

    Article  PubMed  CAS  Google Scholar 

  • Nicholson, J. K., Focall, P. J. D., Spraul, M., Farrant, R. D., & Lindon, J. C. (1995). 750 MHz 1H and 1H–13C NMR spectroscopy of human blood plasma. Analytical Chemistry, 67, 793–811.

    Article  PubMed  CAS  Google Scholar 

  • Park, S. Y., Kim, Y. J., Gao, A. C., Mohler, J. L., Onate, S. A., Hidalgo, A. A., et al. (2006). Hypoxia increases androgen receptor activity in prostate cancer cells. Cancer Research, 66, 5121–5129.

    Article  PubMed  CAS  Google Scholar 

  • Putluri, N., Shojaie, A., Vasu, V. T., Nalluri, S., Vareed, S. K., Putluir, V., et al. (2011). Metabolomic profiling reveals a role for androgen in activating amino acid metabolism and methylation in prostate cancer cells. PLoS ONE, 6, doi:10.1371/journal.pone.0021417.

  • Qi, H., Fillion, C., Labrie, Y., Grenier, J., Fournier, A., Berger, L., et al. (2002). AlbZIP, a novel bZIP gene located on chromosome 1q21.3 that is highly expressed in prostate tumors and of which the expression is up-regulated by androgens in LNCaP human prostate cancer cells. Cancer Research, 62, 721–733.

    PubMed  CAS  Google Scholar 

  • Rantalainen, M., Cloarec, O., Beckonert, O., Wilson, I. D., Jackson, D., Tonge, R., et al. (2006). Statistically integrate metabonomic-proteomic studies on a human prostate cancer xenograft model in mice. Journal of Proteome Research, 5, 2642–2655.

    Article  PubMed  CAS  Google Scholar 

  • Ross, B. D. (1991). Biochemical considerations in 1H spectroscopy. Glutamate and glutamine; Myo-inositol and related metabolites. NMR in Biomedicine, 4, 59–63.

    Article  PubMed  CAS  Google Scholar 

  • Serkova, N. J., Gamito, E. J., Jones, R. H., O’Donnell, C., Brown, J. L., Green, S., et al. (2008). The metabolites citrate, myo-inositol, and spermine are potential age-independent markers of prostate cancer in human expressed prostatic secretions. The Prostate, 68, 620–628.

    Article  PubMed  CAS  Google Scholar 

  • Sitter, B., Sonnewald, U., Spraul, M., Fjösne, H. E., & Gribbestad, I. S. (2002). High-resolution magic angle spinning MRS of breast cancer tissue. NMR in Biomedicine, 15, 327–337.

    Article  PubMed  CAS  Google Scholar 

  • Song, K., Wang, H., Krebs, T. L., Wang, B., Kelley, T. J., & Danielpour, D. (2010). DHT selectively reverses Smad3-mediated/TGF-2-induced responses through transcriptional down-regulation of Smad3 in prostate epithelial cells. Molecular Endocrinology, 24, 2019–2029.

    Article  PubMed  CAS  Google Scholar 

  • Sreekumar, A., Poisson, L. M., Rajendiran, T. M., Khan, A. P., Cao, Q., Yu, J., et al. (2009). Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression. Nature, 457, 910–915.

    Article  PubMed  CAS  Google Scholar 

  • Swanson, M. G., Vigneron, D. B., Tabatabai, Z. L., Males, R. G., Schmitt, L., Carroll, P. R., et al. (2003). Proton HR-MAS spectroscopy and quantitative pathologic analysis of MRI/3D-MRSI-targeted postsurgical prostate tissues. Magnetic Resonance in Medicine, 50, 944–954.

    Article  PubMed  CAS  Google Scholar 

  • Swanson, M. G., Zektzer, A. S., Tabatabia, Z. L., Simko, J., Jarso, S., Keshari, K. R., et al. (2006). Quantitative analysis of prostate metabolites using 1H HR-MAS spectroscopy. Magnetic Resonance in Medicine, 55, 1257–1264.

    Article  PubMed  CAS  Google Scholar 

  • Swanson, M. G., Keshari, K. R., Tabatabai, Z. L., Simko, J. P., Shinohara, K., Carroll, P. R., et al. (2008). Quantification of choline- and ethanolamine-containing metabolites in human prostate tissues using 1H HR-MAS total correlation spectroscopy. Magnetic Resonance in Medicine, 60, 33–40.

    Article  PubMed  CAS  Google Scholar 

  • Teahan, O., Bevan, C. L., Waxman, J., & Keun, H. C. (2010). Metabolic signatures of malignant progression in prostate epithelial cells. International Journal of Biochemistry and Cell Biology,. doi:10.1016/j.biocel.2010.07.003.

    PubMed  Google Scholar 

  • Tessem, M. B., Swanson, M. G., Keshari, K. R., Albers, M. J., Joun, D., Tabatabai, Z. L., et al. (2008). Evaluation of lactate and alanine as metabolic biomarkers of prostate cancer using 1H HR-MAS spectroscopy of biopsy tissues. Magnetic Resonance in Medicine, 60, 510–516.

    Article  PubMed  CAS  Google Scholar 

  • Tiechert, F., Verschoyle, R. D., Greaves, P., Edwards, R. E., Teahan, O., Jones, D. J. L., et al. (2008). Metabolic profiling of transgenic adenocarcinoma of mouse prostate (TRAMP) tissue by 1H-NMR analysis: Evidence for unusual phospholipid metabolism. The Prostate, 68, 1035–1047.

    Article  Google Scholar 

  • Tomlins, A. M., Foxall, P. J. D., Lindon, J. C., Lynch, M. J., Spraul, M., Everett, J. R., et al. (1998). High resolution magic angle spinning 1H nuclear magnetic resonance analysis of intact prostatic hyperplastic and tumour tissues. Analytical Communications, 35, 113–115.

    Article  CAS  Google Scholar 

  • Tomlins, S. A., Rhodes, D. R., Perner, S., Dhanasekaran, S. M., Mehra, R., Sun, X. W., et al. (2005). Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science, 310, 644–648.

    Article  PubMed  CAS  Google Scholar 

  • Tomlins, S. A., Laxman, B., Varambally, S., Cao, X., Yu, J., Helgeson, B. E., et al. (2008). Role of the TMPRSS2-ERG gene fusion in prostate cancer. Neoplasia, 10, 177–188.

    Article  PubMed  CAS  Google Scholar 

  • Van Asten, J. J. A., Cuijpers, V., van de Hulsbergen-Kaa, C., Soede-Huijbregts, C., Witjes, J. A., Verhofstad, A., et al. (2008). High resolution magic angle spinning NMR spectroscopy for metabolic assessment of cancer presence and Gleason score in human prostate needle biopsies. Magnetic Resonance Materials in Physics Biology and Medicine, 21, 435–442.

    Article  Google Scholar 

  • Veselkov, K. A., London, J. C., Ebbels, T. M. D., Crockford, D., Volynkin, V. V., Holmes, E., et al. (2009). Recursive segment-wise peak alignment of biological 1H NMR spectra for improved metabolic biomarker recovery. Analytical Chemistry, 81, 56–66.

    Article  PubMed  CAS  Google Scholar 

  • Wang, Q., Li, W., Zhang, Y., Yuan, X., Xu, K., Yu, J., et al. (2009). Androgen receptor regulates a distinct transcription program in androgen-independent prostate cancer. Cell, 138, 245–256.

    Article  PubMed  CAS  Google Scholar 

  • Wong, J. W. H., Cagney, G., & Cartwright, H. M. (2005a). SpecAlign-processing and alignment of mass spectra datasets. Bioinformatics, 21, 2088–2090.

    Article  PubMed  CAS  Google Scholar 

  • Wong, J. W. H., Durante, C., & Cartwright, H. M. (2005b). Application of fast fourier transform cross-correlation for the alignment of large chromatographic and spectral datasets. Analytical Chemistry, 77, 5655–5661.

    Article  PubMed  CAS  Google Scholar 

  • Wu, C., Taylor, J. L., He, W., Zepeda, A. G., Halperin, E. F., Bielecki, A., et al. (2003). Proton high-resolution magic angle spinning NMR analysis of fresh and previously frozen tissue of human prostate. Magnetic Resonance in Medicine, 50, 1307–1311.

    Article  PubMed  Google Scholar 

  • Yu, J., Yu, J., Mani, R. S., Cao, Q., Brenner, C. J., Cao, X., et al. (2010). An integrated network of androgen receptor, polycomb, and TMPRSS2-ERG gene fusions in prostate cancer progression. Cancer Cell, 17, 443–454.

    Article  PubMed  CAS  Google Scholar 

  • Zhu, H., Mazor, M., Kawano, Y., Walker, M. M., Leung, H. Y., Armstrong, K., et al. (2004). Analysis of Wnt gene expression in prostate cancer: Mutual inhibition of WNT11 and the androgen receptor. Cancer Research, 2004(64), 7918–7926.

    Article  Google Scholar 

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Acknowledgments

We thank Chenomx Inc. (Edmonton, Canada) for providing the Chenomx software. This study was supported by funds from NIH (RR023597 to A.R.) and partly by GM095640 to A.R.).

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Authors and Affiliations

  1. Biophysics and Department of Chemistry, University of Michigan, 930 North University Avenue, Ann Arbor, MI, 48109-1055, USA

    Neil MacKinnon & Ayyalusamy Ramamoorthy

  2. Michigan Center for Translational Pathology, Department of Pathology, University of Michigan, Medical School, Ann Arbor, MI, USA

    Amjad P. Khan, Arul M. Chinnaiyan & Thekkelnaycke M. Rajendiran

  3. Department of Urology, Comprehensive Cancer Center and Howard Hughes Medical Institute, University of Michigan, Medical School, Ann Arbor, MI, USA

    Arul M. Chinnaiyan

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  1. Neil MacKinnon
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  2. Amjad P. Khan
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  3. Arul M. Chinnaiyan
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  4. Thekkelnaycke M. Rajendiran
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  5. Ayyalusamy Ramamoorthy
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Correspondence to Ayyalusamy Ramamoorthy.

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Thekkelnaycke M. Rajendiran and Ayyalusamy Ramamoorthy share senior authorship for this article. Ayyalusamy Ramamooorthy is the corresponding author.

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MacKinnon, N., Khan, A.P., Chinnaiyan, A.M. et al. Androgen receptor activation results in metabolite signatures of an aggressive prostate cancer phenotype: an NMR-based metabonomics study. Metabolomics 8, 1026–1036 (2012). https://doi.org/10.1007/s11306-012-0398-4

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