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High-throughput mutation profiling improves diagnostic stratification of sporadic medullary thyroid carcinomas

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Abstract

Sporadic medullary thyroid carcinoma (MTC) harbors RET gene somatic mutations in up to 50 % of cases, and RAS family gene mutations occur in about 10 %. A timely and comprehensive characterization of molecular alterations is needed to improve MTC diagnostic stratification and design-tailored therapeutic approaches. Twenty surgically resected sporadic MTCs, previously analyzed for RET mutations by Sanger sequencing using DNA from formalin-fixed paraffin-embedded samples, were investigated for intragenic mutations in 50 cancer-associated genes applying a multigene Ion AmpliSeq next-generation sequencing (NGS) technology. Thirteen (65 %) MTCs harbored a RET mutation; 10 were detected at both Sanger and NGS sequencing, while 3 undetected by Sanger were revealed by NGS. One of the 13 RET-mutated cases also showed an F354L germline mutation in STK11. Of the seven RET wild-type MTCs, four cases (57.1 %) harbored a RAS mutation: three in HRAS (all Q61R) and one in KRAS (G12R). The three remaining MTCs (15 %) resulted as wild-type for all the 50 cancer-related genes. Follow-up was available in all but one RET-mutated case. At the end of follow-up, 7 of 12 (58 %) RET-mutated patients had relapsed, while the 4 RAS-mutated MTC patients were disease-free. Two of the three patients with MTC wild-type for all 50 genes relapsed during the follow-up period. Detection of mutations by NGS has the potential to improve the diagnostic stratification of sporadic MTC.

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References

  1. Ciampi R, Mian C, Fugazzola L, Cosci B, Romei C, Barollo S, Cirello V, Bottici V, Marconcini G, Rosa PM, Borrello MG, Basolo F, Ugolini C, Materazzi G, Pinchera A, Elisei R (2013) Evidence of a low prevalence of RAS mutations in a large medullary thyroid cancer series. Thyroid 23:50–57. doi:10.1089/thy.2012.0207

    Article  CAS  PubMed  Google Scholar 

  2. Mian C, Pennelli G, Barollo S, Cavedon E, Nacamulli D, Vianello F, Negro I, Pozza G, Boschin IM, Pelizzo MR, Rugge M, Mantero F, Girelli ME, Opocher G (2011) Combined RET and Ki-67 assessment in sporadic medullary thyroid carcinoma: a useful tool for patient risk stratification. Eur J Endocrinol 164:971–976. doi:10.1530/EJE-11-0079

    Article  CAS  PubMed  Google Scholar 

  3. Siegel R, Naishadham D, Jemal A (2012) Cancer statistics, 2012. CA Cancer J Clin 62:10–29. doi:10.3322/caac.20138

    Article  PubMed  Google Scholar 

  4. Roman S, Lin R, Sosa JA (2006) Prognosis of medullary thyroid carcinoma: demographic, clinical, and pathologic predictors of survival in 1252 cases. Cancer 107:2134–2142. doi:10.1002/cncr.22244

    Article  PubMed  Google Scholar 

  5. de Groot JW, Links TP, Plukker JT, Lips CJ, Hofstra RM (2006) RET as a diagnostic and therapeutic target in sporadic and hereditary endocrine tumors. Endocr Rev 27:535–560. doi:10.1210/er.2006-0017

    Article  PubMed  Google Scholar 

  6. Moura MM, Cavaco BM, Pinto AE, Leite V (2011) High prevalence of RAS mutations in RET-negative sporadic medullary thyroid carcinomas. J Clin Endocrinol Metab 96:E863–E868. doi:10.1210/jc.2010-1921

    Article  CAS  PubMed  Google Scholar 

  7. Agrawal N, Jiao Y, Sausen M, Leary R, Bettegowda C, Roberts NJ, Bhan S, Ho AS, Khan Z, Bishop J, Westra WH, Wood LD, Hruban RH, Tufano RP, Robinson B, Dralle H, Toledo SP, Toledo RA, Morris LG, Ghossein RA, Fagin JA, Chan TA, Velculescu VE, Vogelstein B, Kinzler KW, Papadopoulos N, Nelkin BD, Ball DW (2013) Exomic sequencing of medullary thyroid cancer reveals dominant and mutually exclusive oncogenic mutations in RET and RAS. J Clin Endocrinol Metab 98:E364–E369. doi:10.1210/jc.2012-2703

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Elisei R, Cosci B, Romei C, Bottici V, Renzini G, Molinaro E, Agate L, Vivaldi A, Faviana P, Basolo F, Miccoli P, Berti P, Pacini F, Pinchera A (2008) Prognostic significance of somatic RET oncogene mutations in sporadic medullary thyroid cancer: a 10-year follow-up study. J Clin Endocrinol Metab 93:682–687. doi:10.1210/jc.2007-1714

    Article  CAS  PubMed  Google Scholar 

  9. Barollo S, Pezzani R, Cristiani A, Bertazza L, Rubin B, Bulfone A, Pelizzo MR, Torresan F, Mantero F, Pennelli G, Moro S, Mian C (2013) Functional significance of the novel H-RAS gene mutation M72I in a patient with medullary thyroid cancer. Exp Clin Endocrinol Diabetes 121:546–550. doi:10.1055/s-0033-1351299

    Article  CAS  PubMed  Google Scholar 

  10. Nikiforova MN, Wald AI, Roy S, Durso MB, Nikiforov YE (2013) Targeted next-generation sequencing panel (ThyroSeq) for detection of mutations in thyroid cancer. J Clin Endocrinol Metab 98:E1852–E1860. doi:10.1210/jc.2013-2292

    Article  CAS  PubMed  Google Scholar 

  11. Metzker ML (2010) Sequencing technologies—the next generation. Nat Rev Genet 11:31–46. doi:10.1038/nrg2626

    Article  CAS  PubMed  Google Scholar 

  12. Thomas RK, Nickerson E, Simons JF, Janne PA, Tengs T, Yuza Y, Garraway LA, LaFramboise T, Lee JC, Shah K, O'Neill K, Sasaki H, Lindeman N, Wong KK, Borras AM, Gutmann EJ, Dragnev KH, DeBiasi R, Chen TH, Glatt KA, Greulich H, Desany B, Lubeski CK, Brockman W, Alvarez P, Hutchison SK, Leamon JH, Ronan MT, Turenchalk GS, Egholm M, Sellers WR, Rothberg JM, Meyerson M (2006) Sensitive mutation detection in heterogeneous cancer specimens by massively parallel picoliter reactor sequencing. Nat Med 12:852–855. doi:10.1038/nm1437

    Article  CAS  PubMed  Google Scholar 

  13. Meldrum C, Doyle MA, Tothill RW (2011) Next-generation sequencing for cancer diagnostics: a practical perspective. Clin Biochem Rev 32:177–195

    PubMed Central  PubMed  Google Scholar 

  14. Luchini C, Capelli P, Fassan M, Simbolo M, Mafficini A, Pedica F, Ruzzenente A, Guglielmi A, Corbo V, Scarpa A (2014) Next-generation histopathologic diagnosis: a lesson from a hepatic carcinosarcoma. J Clin Oncol. doi:10.1200/JCO.2012.47.5855

    PubMed  Google Scholar 

  15. Scarpa A, Sikora K, Fassan M, Rachiglio AM, Cappellesso R, Antonello D, Amato E, Mafficini A, Lambiase M, Esposito C, Bria E, Simonato F, Scardoni M, Turri G, Chilosi M, Tortora G, Fassina A, Normanno N (2013) Molecular typing of lung adenocarcinoma on cytological samples using a multigene next generation sequencing panel. PLoS One 8:e80478. doi:10.1371/journal.pone.0080478

    Article  PubMed Central  PubMed  Google Scholar 

  16. Fassan M, Simbolo M, Bria E, Mafficini A, Pilotto S, Capelli P, Bencivenga M, Pecori S, Luchini C, Neves D, Turri G, Vicentini C, Montagna L, Tomezzoli A, Tortora G, Chilosi M, De Manzoni G, Scarpa A (2013) High-throughput mutation profiling identifies novel molecular dysregulation in high-grade intraepithelial neoplasia and early gastric cancers. Gastric Cancer. doi:10.1007/s10120-013-0315-1

    PubMed  Google Scholar 

  17. Simbolo M, Gottardi M, Corbo V, Fassan M, Mafficini A, Malpeli G, Lawlor RT, Scarpa A (2013) DNA qualification workflow for next generation sequencing of histopathological samples. PLoS One 8:e62692. doi:10.1371/journal.pone.0062692

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Zamo A, Bertolaso A, van Raaij AW, Mancini F, Scardoni M, Montresor M, Menestrina F, van Krieken JH, Chilosi M, Groenen PJ, Scarpa A (2012) Application of microfluidic technology to the BIOMED-2 protocol for detection of B-cell clonality. J Mol Diagn 14:30–37. doi:10.1016/j.jmoldx.2011.07.007

    Article  CAS  PubMed  Google Scholar 

  19. Chau NG, Haddad RI (2013) Vandetanib for the treatment of medullary thyroid cancer. Clin Cancer Res: Off J Am Assoc Cancer Res 19:524–529. doi:10.1158/1078-0432.CCR-12-2353

    Article  CAS  Google Scholar 

  20. Elisei R, Schlumberger MJ, Muller SP, Schoffski P, Brose MS, Shah MH, Licitra L, Jarzab B, Medvedev V, Kreissl MC, Niederle B, Cohen EE, Wirth LJ, Ali H, Hessel C, Yaron Y, Ball D, Nelkin B, Sherman SI (2013) Cabozantinib in progressive medullary thyroid cancer. J Clin Oncol 31:3639–3646. doi:10.1200/JCO.2012.48.4659

    Article  CAS  PubMed  Google Scholar 

  21. Haddad RI (2013) How to incorporate new tyrosine kinase inhibitors in the treatment of patients with medullary thyroid cancer. J Clin Oncol 31:3618–3620. doi:10.1200/JCO.2013.51.5098

    Article  CAS  PubMed  Google Scholar 

  22. Viola D, Cappagli V, Elisei R (2013) Cabozantinib (XL184) for the treatment of locally advanced or metastatic progressive medullary thyroid cancer. Future Oncol 9:1083–1092. doi:10.2217/fon.13.128

    Article  CAS  PubMed  Google Scholar 

  23. Wells SA Jr, Robinson BG, Gagel RF, Dralle H, Fagin JA, Santoro M, Baudin E, Elisei R, Jarzab B, Vasselli JR, Read J, Langmuir P, Ryan AJ, Schlumberger MJ (2012) Vandetanib in patients with locally advanced or metastatic medullary thyroid cancer: a randomized, double-blind phase III trial. J Clin Oncol 30:134–141. doi:10.1200/JCO.2011.35.5040

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  24. Forcet C, Etienne-Manneville S, Gaude H, Fournier L, Debilly S, Salmi M, Baas A, Olschwang S, Clevers H, Billaud M (2005) Functional analysis of Peutz-Jeghers mutations reveals that the LKB1 C-terminal region exerts a crucial role in regulating both the AMPK pathway and the cell polarity. Hum Mol Genet 14:1283–1292. doi:10.1093/hmg/ddi139

    Article  CAS  PubMed  Google Scholar 

  25. Boardman LA, Thibodeau SN, Schaid DJ, Lindor NM, McDonnell SK, Burgart LJ, Ahlquist DA, Podratz KC, Pittelkow M, Hartmann LC (1998) Increased risk for cancer in patients with the Peutz-Jeghers syndrome. Ann Intern Med 128:896–899

    Article  CAS  PubMed  Google Scholar 

  26. Spigelman AD, Murday V, Phillips RK (1989) Cancer and the Peutz-Jeghers syndrome. Gut 30:1588–1590

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Yamamoto M, Hoshino H, Onizuka T, Ichikawa M, Kawakubo A, Hayakawa S (1992) Thyroid papillary adenocarcinoma in a woman with Peutz-Jeghers syndrome. Intern Med 31:1117–1119

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work has been supported by the Italian Cancer Genome Project grant from the Italian Ministry of Research (FIRB—RBAP10AHJB).

Conflict of interests

The authors have no competing interests to declare.

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Authors and Affiliations

  1. ARC-Net Research Centre, University and Hospital Trust of Verona, 37134, Verona, Italy

    Michele Simbolo, Matteo Fassan, Andrea Mafficini, Diogo Neves & Aldo Scarpa

  2. Endocrinology Unit, Department of Medicine, University of Padua, 35121, Padua, Italy

    Caterina Mian, Susi Barollo & Elisabetta Cavedon

  3. Department of Pathology and Diagnostics, University and Hospital Trust of Verona, 37134, Verona, Italy

    Matteo Fassan, Maria Scardoni & Aldo Scarpa

  4. Surgical Pathology & Cytopathology Unit, Department of Medicine, University of Padua, 35121, Padua, Italy

    Gianmaria Pennelli & Massimo Rugge

  5. Surgery Unit, Department of Surgical Oncology and Gastroenterology Sciences, University of Padua, 35121, Padua, Italy

    Maria Rosa Pelizzo

  6. Department of Clinical Sciences & Community Health, University of Milan and Endocrine Unit, Fondazione IRCCS CàGranda, Milan, 20122, Italy

    Laura Fugazzola

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  1. Michele Simbolo
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  2. Caterina Mian
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Corresponding author

Correspondence to Matteo Fassan.

Additional information

Michele Simbolo and Caterina Mian shared first authorship.

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Simbolo, M., Mian, C., Barollo, S. et al. High-throughput mutation profiling improves diagnostic stratification of sporadic medullary thyroid carcinomas. Virchows Arch 465, 73–78 (2014). https://doi.org/10.1007/s00428-014-1589-3

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  • DOI: https://doi.org/10.1007/s00428-014-1589-3

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