Abstract
Introduction
Ultrasound examination coupled with fine-needle aspiration (FNA) cytology is the gold standard for the diagnosis of thyroid cancer. However, about 10–40% of these analyses cannot be conclusive on the malignancy of the lesions and lead to surgery. The cytological indeterminate FNA biopsies are mainly constituted of follicular—patterned lesions, which are benign in 80% of the cases.
Objectives
The development of a FNAB classification approach based on the metabolic phenotype of the lesions, complementary to cytology and other molecular tests in order to limit the number of patients undergoing unnecessary thyroidectomy.
Methods
We explored the potential of a NMR-based metabolomics approach to improve the quality of the diagnosis from FNABs, using thyroid tissues collected post-surgically.
Results
The NMR-detected metabolites were used to produce a robust OPLSDA model to discriminate between benign and malignant tumours. Malignancy was correlated with amino acids such as tyrosine, serine, alanine, leucine and phenylalanine and anti-correlated with myo-inositol, scyllo-inositol and citrate. Diagnosis accuracy was of 84.8% when only indeterminate lesions were considered.
Conclusion
These results on model FNAB indicate that there is a clear interest in exploring the possibility to export NMR metabolomics to pre-surgical diagnostics.
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Abbreviations
- FNA:
-
Fine-needle aspiration
- FNAB:
-
Fine-needle aspiration biopsy
- FA:
-
Follicular adenoma
- FTC:
-
Follicular thyroid carcinoma
- PTC:
-
Papillary thyroid carcinoma
- MTC:
-
Medullary thyroid carcinoma
- TCV-PTC:
-
Tll cell variant of papillary thyroid carcinoma
- CV-PTC:
-
Classic variant of papillary thyroid carcinoma
- FV-PTC:
-
Follicular variant of papillary thyroid carcinoma
- HRMAS:
-
High-resolution magic angle spinning
- NMR:
-
Nuclear magnetic resonance
- PCA:
-
Principal component analysis
- OPLS-DA:
-
Orthogonalized projections to the latent structures discriminant analysis
- CV-ANOVA:
-
Cross-validation analysis of variance
- Tg:
-
Thyroglobulin
- TCA:
-
Tricarboxylic acid
- VIP:
-
Variable importance in the prediction
References
Alexander, E. K., Kennedy, G. C., Baloch, Z. W., Cibas, E. S., Chudova, D., Diggans, J., et al. (2012). Preoperative diagnosis of benign thyroid nodules with indeterminate cytology. New England Journal of Medicine, 367(8), 705–715. https://doi.org/10.1056/NEJMoa1203208.
Bauer, D. E., Hatzivassiliou, G., Zhao, F., Andreadis, C., & Thompson, C. B. (2005). ATP citrate lyase is an important component of cell growth and transformation. Oncogene, 24(41), 6314–6322. https://doi.org/10.1038/sj.onc.1208773.
Boroughs, L. K., & DeBerardinis, R. J. (2015). Metabolic pathways promoting cancer cell survival and growth. Nature Cell Biology, 17(4), 351–359. https://doi.org/10.1038/ncb3124.
Cavanagh, J., & Rance, M. (1990). Sensitivity improvement in isotropic mixing (Tocsy) experiments. Journal of Magnetic Resonance, 88(1), 72–85. https://doi.org/10.1016/0022-2364(90)90109-M.
Cibas, E. S., & Ali, S. Z. (2009). The bethesda system for reporting thyroid cytopathology. Thyroid, 19(11), 1159–1165. https://doi.org/10.1089/thy.2009.0274.
Deja, S., Dawiskiba, T., Balcerzak, W., Orczyk-Pawilowicz, M., Glod, M., Pawelka, D., et al. (2013). Follicular adenomas exhibit a unique metabolic profile. (1)H NMR studies of thyroid lesions. PLoS ONE, 8(12), e84637. https://doi.org/10.1371/journal.pone.0084637.
Delgado-Goni, T., Miniotis, M. F., Wantuch, S., Parkes, H. G., Marais, R., Workman, P., et al. (2016). The BRAF inhibitor vemurafenib activates mitochondrial metabolism and inhibits hyperpolarized pyruvate-lactate exchange in BRAF-mutant human melanoma cells. Molecular Cancer Therapeutics, 15, 2987–2999. https://doi.org/10.1158/1535-7163.MCT-16-0068.
Duggal, R., Rajwanshi, A., Gupta, N., & Vasishta, R. K. (2011). Interobserver variability amongst cytopathologists and histopathologists in the diagnosis of neoplastic follicular patterned lesions of thyroid. Diagnostic Cytopathology, 39(4), 235–241. https://doi.org/10.1002/dc.21363.
Elsheikh, T. M., Asa, S. L., Chan, J. K., DeLellis, R. A., Heffess, C. S., LiVolsi, V. A., et al. (2008). Interobserver and intraobserver variation among experts in the diagnosis of thyroid follicular lesions with borderline nuclear features of papillary carcinoma. American Journal of Clinical Pathology, 130(5), 736–744. https://doi.org/10.1309/AJCPKP2QUVN4RCCP.
Franc, B., de la Salmoniere, P., Lange, F., Hoang, C., Louvel, A., de Roquancourt, A., et al. (2003). Interobserver and intraobserver reproducibility in the histopathology of follicular thyroid carcinoma. Human Pathology, 34(11), 1092–1100.
Gharib, H., Papini, E., Paschke, R., Duick, D. S., Valcavi, R., Hegedus, L., et al. (2010). American Association of Clinical Endocrinologists, Associazione Medici Endocrinologi, and European Thyroid Association Medical Guidelines for Clinical Practice for the Diagnosis and Management of Thyroid Nodules. Endocrine Practice, 16(Suppl 1), 1–43. https://doi.org/10.4158/10024.GL.
Gonzalez, H. E., Martinez, J. R., Vargas-Salas, S., Solar, A., Veliz, L., Cruz, F., et al. (2017). A 10-gene classifier for indeterminate thyroid nodules: Development and multicenter accuracy Study. Thyroid, 27(8), 1058–1067. https://doi.org/10.1089/thy.2017.0067.
Guo, S., Qiu, L., Wang, Y., Qin, X., Liu, H., He, M., et al. (2014). Tissue imaging and serum lipidomic profiling for screening potential biomarkers of thyroid tumors by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry. Analytical and Bioanalytical Chemistry, 406(18), 4357–4370. https://doi.org/10.1007/s00216-014-7846-0.
Hatzivassiliou, G., Zhao, F., Bauer, D. E., Andreadis, C., Shaw, A. N., Dhanak, D., et al. (2005). ATP citrate lyase inhibition can suppress tumor cell growth. Cancer Cell, 8(4), 311–321. https://doi.org/10.1016/j.ccr.2005.09.008.
Hodak, S. P., & Rosenthal, D. S., American Thyroid Association Clinical Affairs (2013). Information for clinicians: Commercially available molecular diagnosis testing in the evaluation of thyroid nodule fine-needle aspiration specimens. Thyroid, 23(2), 131–134. https://doi.org/10.1089/thy.2012.0320. C.
Hotelling, H. (1931). The generalization of Student’s ratio. Annals of Mathematical Statistics, 2, 360–378.
Jozwiak, P., Ciesielski, P., Zaczek, A., Lipinska, A., Pomorski, L., Wieczorek, M., et al. (2017). Expression of hypoxia inducible factor 1alpha and 2alpha and its association with vitamin C level in thyroid lesions. Journal of Biomedical Science, 24, 83. https://doi.org/10.1186/s12929-017-0388-y.
Kakudo, K., Kameyama, K., Miyauchi, A., & Nakamura, H. (2014). Introducing the reporting system for thyroid fine-needle aspiration cytology according to the new guidelines of the Japan Thyroid Association. Endocrine Journal, 61(16), 539–552.
Labourier, E., Shifrin, A., Busseniers, A. E., Lupo, M. A., Manganelli, M. L., Andruss, B., et al. (2015). Molecular testing for miRNA, mRNA, and DNA on fine-needle aspiration improves the preoperative diagnosis of thyroid nodules with indeterminate cytology. The Journal of Clinical Endocrinology and Metabolism, 100(7), 2743–2750. https://doi.org/10.1210/jc.2015-1158.
Lloyd, R. V., Osamura, R. Y., Klöppel, G., & Rosai, J. (2017). WHO classification of tumours of endocrine organs. Lyon: IARC.
Lu, J., Hu, S., Miccoli, P., Zeng, Q., Liu, S., Ran, L., et al. (2016). Non-invasive diagnosis of papillary thyroid microcarcinoma: A NMR-based metabolomics approach. Oncotarget, 7(49), 81768–81777. https://doi.org/10.18632/oncotarget.13178.
Matesa, N., Knezevic-Obad, A., Ostovic, K. T., Kardum-Skelin, I., Moslavac, S., Vasilj, A., et al. (2012). Croatian Society for Clinical Cytology guidelines for thyroid cytology. Liječnički vjesnik, 134(7–8), 203–207.
Meiser, J., & Vazquez, A. (2016). Give it or take it: The flux of one-carbon in cancer cells. The FEBS Journal, 283(20), 3695–3704. https://doi.org/10.1111/febs.13731.
Metallo, C. M., Gameiro, P. A., Bell, E. L., Mattaini, K. R., Yang, J., Hiller, K., et al. (2011). Reductive glutamine metabolism by IDH1 mediates lipogenesis under hypoxia. Nature, 481(7381), 380–384. https://doi.org/10.1038/nature10602.
Miccoli, P., Torregrossa, L., Shintu, L., Magalhaes, A., Chandran, J., Tintaru, A., et al. (2012). Metabolomics approach to thyroid nodules: A high-resolution magic-angle spinning nuclear magnetic resonance-based study. Surgery, 152(6), 1118–1124 doi. https://doi.org/10.1016/J.Surg.2012.08.037.
Nardi, F., Basolo, F., Crescenzi, A., Fadda, G., Frasoldati, A., Orlandi, F., et al. (2014). Italian consensus for the classification and reporting of thyroid cytology. Journal of Endocrinological Investigation, 37(6), 593–599. https://doi.org/10.1007/s40618-014-0062-0.
Netea-Maier, R. T., Hunsucker, S. W., Hoevenaars, B. M., Helmke, S. M., Slootweg, P. J., Hermus, A. R., et al. (2008). Discovery and validation of protein abundance differences between follicular thyroid neoplasms. Cancer Research, 68(5), 1572–1580. https://doi.org/10.1158/0008-5472.CAN-07-5020.
Nikiforov, Y. E., Carty, S. E., Chiosea, S. I., Coyne, C., Duvvuri, U., Ferris, R. L., et al. (2015). Impact of the multi-gene ThyroSeq next-generation sequencing assay on cancer diagnosis in thyroid nodules with atypia of undetermined significance/follicular lesion of undetermined significance cytology. Thyroid, 25(11), 1217–1223. https://doi.org/10.1089/thy.2015.0305.
Nordio, M., & Pajalich, R. (2013). Combined treatment with Myo-inositol and selenium ensures euthyroidism in subclinical hypothyroidism patients with autoimmune thyroiditis. J Thyroid Res. https://doi.org/10.1155/2013/424163.
Panebianco, F., Mazzanti, C., Tomei, S., Aretini, P., Franceschi, S., Lessi, F., et al. (2015). The combination of four molecular markers improves thyroid cancer cytologic diagnosis and patient management. BMC Cancer, 15(1), 918. https://doi.org/10.1186/s12885-015-1917-2.
Philippe, I., & Hubert, L. (2016). The reduced concentration of citrate in cancer cells: An indicator of cancer aggressiveness and a possible therapeutic target. Drug Resistance Updates, 29, 47–53. https://doi.org/10.1016/j.drup.2016.09.003.
Poma, A. M., Giannini, R., Piaggi, P., Ugolini, C., Materazzi, G., Miccoli, P., et al. (2018). A six-gene panel to label follicular adenoma, low- and high-risk follicular thyroid carcinoma. Endocrine Connections, 7(1), 124–132. https://doi.org/10.1530/EC-17-0261.
Rinaldi, S., Plummer, M., Biessy, C., Tsilidis, K. K., Ostergaard, J. N., Overvad, K., et al. (2014). Thyroid-stimulating hormone, thyroglobulin, and thyroid hormones and risk of differentiated thyroid carcinoma: The EPIC study. Journal of the National Cancer Institute, 106(6), dju097. https://doi.org/10.1093/jnci/dju097.
Ryoo, I., Kwon, H., Kim, S. C., Jung, S. C., Yeom, J. A., Shin, H. S., et al. (2016). Metabolomic analysis of percutaneous fine-needle aspiration specimens of thyroid nodules: Potential application for the preoperative diagnosis of thyroid cancer. Scientific Reports, 6, 30075. https://doi.org/10.1038/srep30075.
Savorani, F., Tomasi, G., & Engelsen, S. B. (2010). icoshift: A versatile tool for the rapid alignment of 1D NMR spectra. Journal of Magnetic Resonance, 202(2), 190–202. https://doi.org/10.1016/j.jmr.2009.11.012.
Schleucher, J., Schwendinger, M., Sattler, M., Schmidt, P., Schedletzky, O., Glaser, S. J., et al. (1994). A general enhancement scheme in heteronuclear multidimensional NMR employing pulsed field gradients. Journal of Biomolecular NMR, 4(2), 301–306.
Tian, Y., Nie, X., Xu, S., Li, Y., Huang, T., Tang, H., et al. (2015). Integrative metabonomics as potential method for diagnosis of thyroid malignancy. Scientific Reports, 5, 14869. https://doi.org/10.1038/srep14869.
Torregrossa, L., Shintu, L., Chandran, J. N., Tintaru, A., Ugolini, C., Magalhaes, A., et al. (2012). Toward the reliable diagnosis of indeterminate thyroid lesions: A HRMAS NMR-based metabolomics case of study. Journal of Proteome Research, 11(6), 3317–3325 doi. https://doi.org/10.1021/Pr300105e.
Unfer, V., Facchinetti, F., Orru, B., Giordani, B., & Nestler, J. (2017). Myo-inositol effects in women with PCOS: A meta-analysis of randomized controlled trials. Endocrine Connections, 6(8), 647–658. https://doi.org/10.1530/EC-17-0243.
Weber, F., Teresi, R. E., Broelsch, C. E., Frilling, A., & Eng, C. (2006). A limited set of human MicroRNA is deregulated in follicular thyroid carcinoma. The Journal of Clinical Endocrinology and Metabolism, 91(9), 3584–3591. https://doi.org/10.1210/jc.2006-0693.
Wishart, D. S., Jewison, T., Guo, A. C., Wilson, M., Knox, C., Liu, Y., et al. (2013). HMDB 3.0–the human metabolome database in 2013. Nucleic Acids Research, 41(D1), D801–D807. https://doi.org/10.1093/nar/gks1065.
Wojtowicz, W., Zabek, A., Deja, S., Dawiskiba, T., Pawelka, D., Glod, M., et al. (2017). Serum and urine (1)H NMR-based metabolomics in the diagnosis of selected thyroid diseases. Scientific Reports, 7(1), 9108. https://doi.org/10.1038/s41598-017-09203-3.
Xu, Y. N., Zheng, X. J., Qiu, Y. P., Jia, W., Wang, J. D., & Yin, S. K. (2015). Distinct metabolomic profiles of papillary thyroid carcinoma and benign thyroid adenoma. Journal of Proteome Research, 14(8), 3315–3321. https://doi.org/10.1021/acs.jproteome.5b00351.
Yao, Z., Yin, P., Su, D., Peng, Z., Zhou, L., Ma, L., et al. (2011). Serum metabolic profiling and features of papillary thyroid carcinoma and nodular goiter. Molecular Biosystems, 7(9), 2608–2614. https://doi.org/10.1039/c1mb05029j.
Acknowledgements
This study was funded by Agence nationale de la recherche (Grant No. ANR-2011- JS08-014-01).
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PM, FB and SC designed research. LR, AS performed research. AS and LT performed the cytological and histological analyses. LS analysed the data. LS and LT wrote the paper. All authors read and approved the manuscript.
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All procedures performed in studies involving human participants were in accordance with the 1964 Helsinki declaration and its later amendments and approved by the Local Ethics Committee of the Azienda Ospedaliero Universitaria Pisana.
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Rezig, L., Servadio, A., Torregrossa, L. et al. Diagnosis of post-surgical fine-needle aspiration biopsies of thyroid lesions with indeterminate cytology using HRMAS NMR-based metabolomics. Metabolomics 14, 141 (2018). https://doi.org/10.1007/s11306-018-1437-6
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DOI: https://doi.org/10.1007/s11306-018-1437-6