Abstract
HR-MAS NMR spectroscopy was employed to monitor the metabolic profiles of Modena BioBank breast samples over 1 year of freezing at −80 °C. The study includes 22 adult female patients living in Modena and its hinterland, who underwent total mastectomy or quadrantectomy in 2011–2012. Variations occur, especially affecting phosphocholine (PC) and choline. This is not a trivial finding, since many studies base the distinction between neoplastic and healthy tissues or the assessment of tumor grade on the analysis of choline metabolites. Despite the changes observed, we established that the diagnostic power of the HR-MAS NMR spectra of frozen samples is preserved, at least as far as the distinction between neoplastic and healthy samples is concerned. Lactate (Lac), PC, phosphoethanolamine (PE), taurine (Tau), myo-inositol (Myo) and glucose (Glc) are biomarkers that can be used to distinguish healthy from neoplastic tissues, whereas some metabolite ratios, such as Lac + PE + Tau/Glc + Myo, seem to have even higher discrimination potential.
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Abbreviations
- Asc:
-
Ascorbate
- TGgl :
-
Bonded glycerol
- CPMG:
-
Carr–Purcell–Meiboom–Gill
- Cho:
-
Choline
- ChoCC:
-
Choline containing compounds
- E:
-
Ethanolamine
- Glc:
-
Glucose
- GPC:
-
Glycerophosphocholine
- Gly:
-
Glycine
- HR-MAS:
-
High-Resolution Magic Angle Spinning
- Lac:
-
Lactate
- Myo:
-
Myo-inositol
- NMR:
-
Nuclear Magnetic Resonance
- PLS-DA:
-
Partial Least Squares-Discriminant Analysis
- PC:
-
Phosphocholine
- PE:
-
Phosphoethanolamine
- PCA:
-
Principal Component Analysis
- ROCCET:
-
Receiver Operating Characteristic Curve Explorer & Tester
- Tau:
-
Taurine
References
Ackerstaff, E., Glunde, K., & Bhujwalla, Z. M. (2003). Choline phospholipid metabolism: A target in cancer cells? Journal of Cellular Biochemistry, 90, 525–533.
Aoyama, C., Liao, H., & Ishidate, K. (2004). Structure and function of choline kinase isoforms in mammalian cells. Progress in Lipid Research, 43, 266–281.
Bathen, F. T., Geurts, B., Sitter, B., Fjøsne, H. E., Lundgren, S., Buydens, L. M., et al. (2013). Feasibility of MR metabolomics for immediate analysis of resection margins during breast cancer surgery. PLoS ONE, 8, e61578.
Bathen, F. T., Sitter, B., Sjøbakk, T. E., Tessem, M. B., & Gribbestad, I. S. (2010). Magnetic resonance metabolomics of intact tissue: A biotechnological tool in cancer diagnostics and treatment evaluation. Cancer Research, 70, 6692–6696.
Bax, A. (1985). A spatially selective composite 90° radiofrequency pulse. Journal of Magnetic Resonance, 65, 142–145.
Brizel, D. M., Schroeder, T., Scher, R. L., Walenta, S., Clough, R. W., Dewhirst, M. W., et al. (2001). Elevated tumor lactate concentrations predict for an increased risk of metastases in head-and-neck cancer. International Journal of Radiation Oncology Biology Physics, 51, 349–353.
Cacciatore, S., Hu, X., Viertler, C., Kap, M., Bernhardt, G. A., Mischinger, H. J., et al. (2013). Effects of intra- and post-operative ischemia on the metabolic profile of clinical liver tissue specimens monitored by NMR. Journal of Proteome Research, 12, 5723–5729.
Cao, M. D., Sitter, B., Bathen, T. F., Bofin, A., Lønning, P. E., Lundgren, S., et al. (2012). Predicting long-term survival and treatment response in breast cancer patients receiving neoadjuvant chemotherapy by MR metabolic profiling. NMR in Biomedicine, 25, 369–378.
Cheng, L. L., Chang, I. W., Smith, B. L., & Gonzalez, R. G. (1998). Evaluating human breast ductal carcinomas with high-resolution magic-angle spinning proton magnetic resonance spectroscopy. Journal of Magnetic Resonance, 135, 194–202.
Choi, J. S., Baek, H. M., Kim, S., Kim, J. M., Youk, H. J., Moon, H. J., et al. (2012). HR-MAS MR spectroscopy of breast cancer tissue obtained with core needle biopsy: Correlation with prognostic factors. PLoS ONE, 7, e51712.
Elston, C. W., & Ellis, I. O. (1991). Pathologic prognostic factors in breast cancer. The value of histological grades in breast cancer. Experience from a large study with long-term follow-up. Histopathology, 19, 403–410.
Esmaeili, M., Moestue, S. A., Hamans, B. C., Veltien, A., Kristian, A., Engebraten, O., et al. (2014). In vivo 31P magnetic resonance spectroscopic imaging (MRSI) for metabolic profiling of human breast cancer xenografts. Journal of Magnetic Resonance Imaging. doi:10.1002/jmri.24588.
Federico, M., Artioli, M. E., Braghiroli, B., Cirilli, C., Iachetta, F., Luminari, S., et al. (2013). Regione Emilia Romagna. I tumori nelle province di Modena e Parma; Rapporto novembre 2013. Registro tumori della Provincia di Modena and Registro tumori della Provincia di Parma, 32, 6–7.
Ferlay, J., Soerjomataram, I., Ervik, M., Dikshit, R., Eser, S., Mathers, C., et al. (2013). GLOBOCAN 2012v1.0, Cancer Incidence and mortality Worldwide: IARC CancerBase No 11. Lyon, France: International Agency for Research on Cancer
Ferreira, A. K., Meneguelo, R., Pereira, A., Mendonca, O., Filho, R., Chierice, G. O., et al. (2013). Synthetic phosphoethanolamine induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells through the mitochondrial pathway. Biomedicine & Pharmacotherapy, 67, 481–487.
Glunde, K., Bhujwalla, Z. M., & Ronen, S. M. (2011). Choline metabolism in malignant transformation. Nature Reviews Cancer, 11, 835–848.
Jain, M., Nilsson, R., Sharma, S., Madhusudhan, N., Kitami, T., Souza, A. L., et al. (2012). Metabolite profiling identifies a key role for glycine in rapid cancer cell proliferation. Science, 336, 1040–1044.
Jiang, L., Greenwood, T. R., Artemov, D., Raman, V., Winnard, P. T., Heeren, R. M. A., et al. (2012). Localized hypoxia results in spatially heterogeneous metabolic signatures in breast tumor models. Neoplasia, 14, 732–741.
Jordan, K. W., He, W., Halpern, E. F., Wu, C. L., & Cheng, L. L. (2007). Evaluation of tissue metabolites with high resolution magic angle spinning MR spectroscopy human prostate samples after 3-year storage at −80 °C. Biomarker Insights, 2, 147–154.
Kinross, J. M., Holmes, E., Darzi, A. W., & Nicholson, J. K. (2011). Metabolic phenotyping for monitoring surgical patients. Lancet, 377, 1817–1819.
Li, M., Song, Y., Cho, N., Chang, J. M., Koo, H. R., Yi, A., et al. (2011). An HR-MAS MR metabolomics study on breast tissues obtained with core needle biopsy. PLoS ONE, 6, e25563.
Meiboom, S., & Gill, D. (1958). Modified spin-echo method for measuring nuclear relaxation time. Review of Scientific Instruments, 20, 688–691.
Moestue, S., Sitter, B., Bathern, T. F., Tessem, M. B., & Gribbestad, I. S. (2011). HR MAS MR spectroscopy in metabolic characterization of cancer. Current Topics in Medicinal Chemistry, 11, 2–26.
Opstad, K. S., Bell, B. A., Griffiths, J. R., & Howe, F. A. (2008). An investigation of human brain tumour lipids by high-resolution magic angle spinning 1H MRS and histological analysis. NMR in Biomedicine, 21, 1138–1147.
Peeling, J., & Sutherland, G. (1992). High-resolution 1H NMR spectroscopy studies of extracts of human cerebral neoplasms. Magnetic Resonance in Medicine, 24, 123–136.
Piotto, M., Moussallieh, F. M., Neuville, A., Bellocq, J. P., Elbayed, K., & Namer, I. J. (2012). Towards real-time metabolic profiling of a biopsy specimen during a surgical operation by 1H high resolution magic angle spinning nuclear magnetic resonance: A case report. Journal of Medical Case Reports, 6, 22.
Podo, F. (1999). Tumour phospholipid metabolism. NMR in Biomedicine, 12, 413–439.
Quaglia, A., Lillini, R., Crocetti, E., Buzzoni, C., Vercelli, M., & AIRTUM Working Group. (2013). Incidence and mortality trends for four major cancers in the elderly and middle-aged adults: An international comparison. Surgical Oncology, 22, e31–e38.
Quennet, V., Yaromina, A., Zips, D., Rosner, A., Walenta, S., Baumann, M., et al. (2006). Tumor lactate content predicts for response to fractionated irradiation of human squamous cell carcinomas in nude mice. Radiotherapy and Oncology, 81, 130–135.
Rossi, S., Crocetti, E., Capocaccia, R., Gatta, G., & AIRTUM Working Group. (2013). Estimates of cancer burden in Italy. Tumori, 99, 416–424.
Sitter, B., Lundgren, S., Bathen, T. F., Halgunset, J., Fjosne, H. E., & Gribbestad, I. S. (2006). Comparison of HR MAS MR spectroscopic profiles of breast cancer tissue with clinical parameters. NMR in Biomedicine, 19, 30–40.
Sitter, B., Sonnewald, U., Spraul, M., Fjosne, H. E., & Gribbestad, I. S. (2002). High-resolution magic angle spinning MRS of breast cancer tissue. NMR in Biomedicine, 15, 327–337.
Sobin, L. H., Gospodarowicz, M. K., & Wittekind, Ch. (2009). TNM classification of malignant tumors (7th ed.). Oxford: Wiley-Blackwell.
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.
Tavassoli, F. A., & Devilee, P. (Eds.). (2003). Word Health Organization: Tumor of the breast and female genital organ. Oxford: Oxford University Press.
Tripathi, P., Kamarajan, P., Somashekar, B. S., MacKinnon, N., Chinnaiyan, A. M., Kapila, Y. L., et al. (2012). Delineating metabolic signatures of head and neck squamous cell carcinoma: Phospholipase A2, a potential therapeutic target. International Journal of Biochemistry & Cell Biology, 44, 1852–1861.
Tripathi, P., Somashekar, B. S., Ponnusamy, M., Gursky, A., Dailey, S., Kunju, P., et al. (2013). HR-MAS NMR tissue metabolomic signatures cross-validated by mass spectrometry distinguish bladder cancer from benign disease. Journal of Proteome Research, 12, 3519–3528.
Vander Heiden, M. G., Cantley, L. C., & Thompson, C. B. (2009). Understanding the Warburg effect: The metabolic requirements of cell proliferation. Science, 324, 1029–1033.
Walenta, S., Wetterling, M., Lehrke, M., Schwickert, G., Sundfør, K., Rofstad, E. K., et al. (2000). High lactate levels predict likelihood of metastases, tumor recurrence, and restricted patient survival in human cervical cancers. Cancer Research, 60, 916–921.
Warburg, O. (1956). On the origin of cancer cells. Science, 123, 309–314.
Xia, J., Broadhurst, D., Wilson, M., & Wishart, D. S. (2012a). Translational biomarker discovery in clinical metabolomics: An introductory tutorial. Metabolomics, 9, 280–299.
Xia, J., Mandal, R., Sinelnikov, I., Broadhurst, D., & Wishart, D. S. (2012b). MetaboAnalyst 2.0—a comprehensive server for metabolomic data analysis. Nucleic Acids Research, 40, W127–W133.
Xia, J., Psychogios, N., Young, N., & Wishart, D. S. (2009). MetaboAnalyst: A web server for metabolomic data analysis and interpretation. Nucleic Acids Research, 37, W652–W660.
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Righi, V., Schenetti, L., Maiorana, A. et al. Assessment of freezing effects and diagnostic potential of BioBank healthy and neoplastic breast tissues through HR-MAS NMR spectroscopy. Metabolomics 11, 487–498 (2015). https://doi.org/10.1007/s11306-014-0709-z
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DOI: https://doi.org/10.1007/s11306-014-0709-z