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Monitoring liver alterations during hepatic tumorigenesis by NMR profiling and pattern recognition

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Abstract

Human hepatocellular carcinoma (HCC) is the most recurrent malignancy of the liver and represents one of the main causes of cancer death worldwide. Furthermore, the liver is the most frequent site of metastatic colonization, and hepatic metastases are far more common than primary cancers in Western countries. A possible way of investigating liver diseases is to study the tissue metabolic profiles. High-resolution nuclear magnetic resonance (NMR) spectroscopy of hepatic tissue extracts was combined with pattern-recognition and visualization techniques to uncover metabolic differences among analyzed tissue types. Extracts were from primary HCC, chronic hepatitis-C-virus related cirrhotic tissues, hepatic metastases from colorectal carcinomas, and non-cirrhotic normal liver tissues adjacent to metastases as controls. We identified all metabolites present in the NMR spectra, and after statistical evaluation of all spectral classes, we were able to define the metabolic changes underlying the different liver conditions and diseases. In particular, the lactate and the glucose tissue signals were found to primarily discriminate the different histological samples. We followed the biochemical changes of human hepatic lesions through primary (HCC) and secondary (metastases from colorectal carcinoma) liver tumors, cirrhotic tissues, and non-cirrhotic histologically-confirmed normal liver tissues adjacent to metastases, achieving a metabolic differentiation of the various pathological states based upon the variation of the intracellular lactate/glucose ratio. It is suggested that such a signal pattern may act as a potential marker for assessing pathological hepatic lesions.

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References

  • Atalay, G., Biganzoli, L., Renard, F., et al. (2003). Clinical outcome of breast cancer patients with liver metastases alone in the anthracycline-taxane era: A retrospective analysis of two prospective, randomized metastatic breast cancer trials. European Journal of Cancer, 39, 2439–2449.

    Article  CAS  PubMed  Google Scholar 

  • Bax, A., & Davis, D. G. (1985). MLEV-17-based two-dimensional homo-nuclear magnetization transfer spectroscopy. Journal of Magnetic Resonance, 65, 355–360.

    CAS  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Chen, Y. K., Hsieh, D. S., Liao, C. S., et al. (2005). Utility of FDG-PET for investigating unexplained serum AFP elevation in patients with suspected hepatocellular carcinoma recurrence. Anticancer Research, 25, 4719–4725.

    PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Chhieng, D. C. (2004). Fine needle aspiration biopsy of liver—an update. World Journal of Surgical Oncology, 2. http://www.wjso.com/content/2/1/5.

  • Cho, S. G., Kim, M. Y., Kim, H. J., et al. (2003). Chronic hepatitis: In vivo proton MR spectroscopic evaluation of the liver and correlation with histopathologic findings. Radiology, 226, 288–289.

    Article  Google Scholar 

  • Christofori, G. (2006). New signals from the invasive front. Nature, 441, 444–450.

    Article  CAS  PubMed  Google Scholar 

  • Cline, G. W., Jucker, B. M., Trajanoski, Z., Rennings, A. J., & Shulman, G. I. (1998). A novel 13C NMR method to assess intracellular glucose concentration in muscle, in vivo. American Journal of Physiology, 274, 381–389.

    Google Scholar 

  • Curado, M. P., Edwards, B., Shin, H. R., et al. (2007). Cancer incidence in five continents (Vol. IX). Lyon: International Agency for Research on Cancer (IARC) Scientific Publications, No. 160.

  • Eriksson, L., Johansson, E., Kettaneh-Wold, N., Trygg, J., Wikström, C., & Wold, S. (2006). Multi- and megavariate data analysis. Basic principles and applications. Umeå: Umetrics AB.

    Google Scholar 

  • Eriksson, L., Johansson, E., Kettaneh-Wold, N., & Wold, S. (1999). Introduction to multi and megavariate data analysis using projection methods (PCA and PLS-DA). Malmö: UMETRICS Academy Publisher.

    Google Scholar 

  • Fan, T. W. M. (1996). Metabolite profiling by one- and two-dimensional NMR analysis of complex mixtures. Progress in Nuclear Magnetic Resonance Spectroscopy, 28, 161–219.

    CAS  Google Scholar 

  • Foley, L. M., Towner, R. A., & Painter, D. M. (2001). In vivo image-guided 1H-magnetic resonance spectroscopy of the serial development of hepatocarcinogenesis in an experimental animal model. Biochimica et Biophysica Acta, 1526, 230–236.

    CAS  PubMed  Google Scholar 

  • Gatenby, R. A., & Gillies, R. J. (2004). Why do cancers have high aerobic glycolysis? Nature Reviews Cancer, 4, 891–899.

    Article  CAS  PubMed  Google Scholar 

  • Griesinger, C., Otting, G., Wüthrich, K., & Ernst, R. R. (1988). Clean TOCSY for proton spin system identification in macromolecules. Journal of the American Chemical Society, 110, 7870–7872.

    Article  CAS  Google Scholar 

  • Ho, C. L., Chen, S., Yeung, D. W. C., & Cheng, T. K. C. (2007). Dual-tracer PET/CT imaging in evaluation of metastatic hepatocellular carcinoma. Journal of Nuclear Medicine, 48, 902–909.

    Article  CAS  PubMed  Google Scholar 

  • Hockel, M., & Vaupel, P. (2001). Tumor hypoxia: Definitions and current clinical, biologic, and molecular aspects. Journal of the National Cancer Institute, 93, 266–276.

    Article  CAS  PubMed  Google Scholar 

  • Hwang, T.-L., & Shaka, A. J. (1995). Water suppression that works: Excitation sculpting using arbitrary waveforms and pulse field gradients. Journal of Magnetic Resonance, 112, 275–279.

    CAS  Google Scholar 

  • Jonas, S., Bechstein, W. O., Steinüller, T., et al. (2001). Vascular invasion and histopathologic grading determine outcome after liver transplantation for hepatocellular carcinoma in cirrhosis. Hepatology, 33, 1080–1086.

    Article  CAS  PubMed  Google Scholar 

  • Kashiwagi, T. (2004). FDG-PET and hepatocellular carcinoma. Journal of Gastroenterology, 39, 1017–1018.

    Article  PubMed  Google Scholar 

  • Kay, L. E., Keifer, P., & Saarinen, T. (1992). Pure absorption gradient enhanced heteronuclear single quantum correlation spectroscopy with improved sensitivity. Journal of Magnetic Resonance, 114, 10663–10665.

    CAS  Google Scholar 

  • Le Belle, J. E., Harris, N. G., Williams, S. R., & Bhakoo, K. K. (2002). A comparison of cell and tissue extraction techniques using high-resolution 1H-NMR spectroscopy. NMR in Biomedicine, 15, 37–44.

    Article  CAS  PubMed  Google Scholar 

  • Lin, C. Y., Wu, H., Tjeerdema, R. S., & Viant, M. R. (2007). Evaluation of metabolite extraction strategies from tissue samples using NMR metabolomics. Metabolomics, 3, 55–67.

    Article  CAS  Google Scholar 

  • Lindon, J. C., Holmes, E., & Nicholson, J. K. (2001). Pattern recognition methods and applications in biomedical magnetic resonance. Progress in Nuclear Magnetic Resonance Spectroscopy, 39, 1–40.

    Article  CAS  Google Scholar 

  • Lindon, J. C., Nicholson, J. K., Holmes, E., et al. (2005). Standard metabolic reporting structures working group. Summary recommendations for standardization and reporting of metabolic analyses. Nature Biotechnology, 23, 833–838.

    Article  CAS  PubMed  Google Scholar 

  • London, R. E. (1988). 13C labelling in studies of metabolic regulation. Progress in Nuclear Magnetic Resonance Spectroscopy, 20, 337–383.

    Article  CAS  Google Scholar 

  • Mountford, C. E., Doran, S., Lean, C. L., & Russell, P. (2004). Proton MRS can determine the pathology of human cancers with a high level of accuracy. Chemical Reviews, 104, 3677–3704.

    Article  CAS  PubMed  Google Scholar 

  • Mueller, G. C., Hussein, H. K., Carlos, R. C., Nghiem, H. V., & Francis, I. R. (2003). Effectiveness of MR imaging in characterizing small hepatic lesions: Routine versus export interpretation. American Journal of Roentgenology, 180, 673–680.

    PubMed  Google Scholar 

  • Nicholson, J. K., Connelly, J., Lindon, J. C., & Holmes, E. (2002). Metabonomics: A platform for studying drug toxicity and gene function. Nature Reviews Drug Discovery, 1, 153–162.

    Article  CAS  PubMed  Google Scholar 

  • Nicholson, J. K., Lindon, J. C., & Holmes, E. (1999). Metabonomics: Understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica, 29, 1181–1189.

    Article  CAS  PubMed  Google Scholar 

  • Okazumi, S., Isono, K., Enomoto, K., et al. (1992). Evaluation of liver tumors using fluorine-18-fluorodeoxyglucose PET: Characterization of tumor and assessment of effect of treatment. Journal of Nuclear Medicine, 33, 333–339.

    CAS  PubMed  Google Scholar 

  • Palmer, A. G., I. I. I., Cavanagh, J., Wright, P. E., & Rance, M. (1991). Sensitivity improvement in proton detected heteronuclear correlation experiments. Journal of Magnetic Resonance, 93, 151–170.

    CAS  Google Scholar 

  • Park, J.-W., Kim, J. H., Kim, S. K., et al. (2008). A prospective evaluation of 18F-FDG and 11C-acetate PET/CT for detection of primary and metastatic hepatocellular carcinoma. Journal of Nuclear Medicine, 49, 1912–1921.

    Article  PubMed  Google Scholar 

  • Patterson, S. A., Khalil, H. I., & Panicek, D. M. (2006). MRI evaluation of small hepatic lesions in women with breast cancer. American Journal of Roentgenology, 187, 307–312.

    Article  PubMed  Google Scholar 

  • Quennet, V., Yaromina, A., Zips, D., et al. (2006). Tumor lactate content predicts for response to fractionated irradiation of human squamous cell carcinomas in nude mice. Radiotherapy & Oncology, 81, 130–135.

    Article  CAS  Google Scholar 

  • Seenu, V., Kumar, M., Sharma, U., Datta Gupta, S., Metha, S. N., & Jagannathan, N. R. (2005). Potential of magnetic resonance spectroscopy to detect metastasis in axillary lymph nodes in breast cancer. Magnetic Resonance Imaging, 23, 1005–1010.

    Article  PubMed  Google Scholar 

  • Soper, R., Himmelreich, U., Painter, D., et al. (2002). Pathology of hepatocellular carcinoma and its precursors using proton magnetic resonance spectroscopy and a statistical classification strategy. Pathology, 34, 417–422.

    Article  PubMed  Google Scholar 

  • Thomas, M., & Zhu, A. (2005). Hepatocellular carcinoma: The need for progress. Journal of Clinical Oncology, 23, 2892–2899.

    Article  PubMed  Google Scholar 

  • Torizuka, T., Tamaki, N., Inokuma, T., et al. (1995). In vivo assessment of glucose metabolism in hepatocellular carcinoma with FDG-PET. Journal of Nuclear Medicine, 36, 1811–1817.

    CAS  PubMed  Google Scholar 

  • Trygg, J., & Wold, S. (2003). O2-PLS, a two-block (X-Y) latent variable regression (LVR) method with an integral OSC filter. Journal of Chemometrics, 17, 53–64.

    Article  CAS  Google Scholar 

  • Usenius, J. P., Kauppinen, R. A., Vainio, P. A., et al. (1994). Quantitative metabolite patterns of human brain tumors: Detection by 1H NMR spectroscopy in vivo and in vitro. Journal of Computer Assisted Tomography, 18, 705–713.

    Article  CAS  PubMed  Google Scholar 

  • Viant, M. R. (2003). Improved methods for the acquisition and interpretation of NMR metabolomic data. Biochemical and Biophysical Research Communications, 310, 943–948.

    Article  CAS  PubMed  Google Scholar 

  • Walenta, S., Schroeder, T., & Mueller-Klieser, W. (2004). Lactate in solid malignant tumors: Potential basis of a metabolic classification in clinical oncology. Current Medicinal Chemistry, 11, 2195–2204.

    CAS  PubMed  Google Scholar 

  • Warburg, O. (1956). On the origin of cancer cells. Science, 123, 309–314.

    Article  CAS  PubMed  Google Scholar 

  • Whitefead, T. L., & Kieber-Emmons, T. (2005). Applying in vitro NMR spectroscopy and 1H NMR metabonomics to breast cancer characterization and detection. Progress in Nuclear Magnetic Resonance Spectroscopy, 47, 165–174.

    Article  Google Scholar 

  • Wiklund, S., Johansson, E., Sjöström, L., et al. (2008). Visualization of GC/TOF-MS-based metabolomics data for identification of biochemically interesting compounds using OPLS class models. Analytical Chemistry, 80, 115–122.

    Article  CAS  PubMed  Google Scholar 

  • Yang, Y., Li, C., Nie, X., et al. (2007). Metabonomic studies of human hepatocellular carcinoma using high-resolution magic-angle spinning 1H NMR spectroscopy in conjunction with multivariate data analysis. Journal of Proteome Research, 6, 2605–2614.

    Article  CAS  PubMed  Google Scholar 

  • Zu, X. L., & Guppy, M. (2004). Cancer metabolism: Facts, fantasy, and fiction. Biochemical and Biophysical Research Communications, 313, 459–465.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We are grateful to patients for their important contribution to this study. We thank Dr. Lorenza Broccardo (S-IN Soluzioni Informatiche, Vicenza, Italy) for helpful discussions, and Emilio P. Castelluccio (ICB-CNR, Pozzuoli) for computer maintenance. Partial financial support was received from the Italian Ministry of Health “Progetto Ricerca Finalizzata”, Italian Association for Cancer Research (AIRC), Sardinia Regional Government “Regione Autonoma della Sardegna”, and Consiglio Nazionale delle Ricerche.

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Authors and Affiliations

  1. Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, 80078, Pozzuoli, NA, Italy

    Debora Paris, Dominique Melck & Andrea Motta

  2. S-IN Soluzioni Informatiche, Via G. Salvemini 9, 36100, Vicenza, Italy

    Matteo Stocchero

  3. Dipartimento di Scienze Biomediche, Università degli Studi di Foggia, 71100, Foggia, Italy

    Oceania D’Apolito & Gaetano Corso

  4. Istituto Nazionale per lo Studio e la Cura dei Tumori, Fondazione G. Pascale, 80131, Naples, Italy

    Rosa Calemma, Giuseppe Castello & Francesco Izzo

  5. Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, 07100, Li Punti-Sassari, SS, Italy

    Giuseppe Palmieri

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  1. Debora Paris
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  2. Dominique Melck
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  4. Oceania D’Apolito
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  6. Giuseppe Castello
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  7. Francesco Izzo
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  9. Gaetano Corso
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  10. Andrea Motta
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Correspondence to Andrea Motta.

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Debora Paris, Dominique Melck and Matteo Stocchero contributed equally to this work.

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Paris, D., Melck, D., Stocchero, M. et al. Monitoring liver alterations during hepatic tumorigenesis by NMR profiling and pattern recognition. Metabolomics 6, 405–416 (2010). https://doi.org/10.1007/s11306-010-0209-8

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