Identification of BRAF V600E mutation in odontogenic tumors by high-performance MALDI-TOF analysis

Abstract

Odontogenic tumors are rare lesions with unknown etiopathogenesis. Most of them are benign, but local aggressiveness, infiltrative potential, and high recurrence rate characterize some entities. The MAP-kinase pathway activation can represent a primary critical event in odontogenic tumorigenesis. Especially, the BRAF V600E mutation has been involved in 80–90% of ameloblastic lesions, offering a biological rationale for developing new targeted therapies. The study aims to evaluate the BRAF V600E mutation in odontogenic lesions, comparing three different detection methods and focusing on the Sequenom MassARRAY System. 81 surgical samples of odontogenic lesions were subjected to immunohistochemical analysis, Sanger Sequencing, and Matrix-Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry (Sequenom). The BRAF V600E mutation was revealed only in ameloblastoma samples. Moreover, the presence of BRAF V600E was significantly associated with the mandibular site (ρ = 0.627; P value <0.001) and the unicystic histotype (ρ = 0.299, P value <0.001). However, any significant difference of 10-years disease-free survival time was not revealed. Finally, Sequenom showed to be a 100% sensitive and 98.1% specific, suggesting its high-performance diagnostic accuracy. These results suggest the MAP-kinase pathway could contribute to ameloblastic tumorigenesis. Moreover, they could indicate the anatomical specificity of the driving mutations of mandibular ameloblastomas, providing a biological rational for developing new targeted therapies. Finally, the high diagnostic accuracy of Sequenom was confirmed.

References

[1] Mascitti M, . Odontogenic tumours: a 25-year epidemiological study in the Marche region of Italy. Eur. Arch. Otorhinolaryngol., 2020, 277: 527-538.

[2] Barnes, L., UniversitèatsSpital Zèurich. Departement Pathologie., International Academy of Pathology., World Health Organization. & International Agency for Research on Cancer. Pathology and Genetics of Head and Neck Tumours (IARC Press, 2005).

[3] Rubini C, . Odontogenic tumors: a retrospective clinicopathological study from two Italian centers. Pathologica, 2017, 109: 35-46.

[4] El-Naggar, A. K. et al. WHO classification of Head and Neck Tumours (International Agency for Research on Cancer, 2017).

[5] Hayward JR. Recurrent ameloblastoma 30 years after surgical treatment. J. Oral. Surg., 1973, 31: 368-370.

[6] Al-Moraissi EA, . What surgical treatment has the lowest recurrence rate following the management of keratocystic odontogenic tumor?: a large systematic review and meta-analysis. J. Craniomaxillofac Surg., 2017, 45: 131-144.

[7] Chae MP, Smoll NR, Hunter-Smith DJ, Rozen WM. Establishing the natural history and growth rate of ameloblastoma with implications for management: systematic review and meta-analysis. PLoS ONE, 2015, 10: e0117241.

[8] Lo Muzio L, . p63 expression correlates with pathological features and biological behaviour of odontogenic tumours. Histopathology, 2006, 49: 211-214.

[9] Diniz MG, Gomes CC, de Sousa SF, Xavier GM, Gomez RS. Oncogenic signalling pathways in benign odontogenic cysts and tumours. Oral. Oncol., 2017, 72: 165-173.

[10] Brown NA, Betz BL. Ameloblastoma: a review of recent molecular pathogenetic discoveries. Biomark. Cancer, 2015, 7: 19-24.

[11] Diniz MG, . DNA damage response activation and cell cycle dysregulation in infiltrative ameloblastomas: a proposed model for ameloblastoma tumor evolution. Exp. Mol. Pathol., 2017, 102: 391-395.

[12] You Z, Liu SP, Du J, Wu YH, Zhang SZ. Advancements in MAPK signaling pathways and MAPK-targeted therapies for ameloblastoma: a review. J. Oral. Pathol. Med, 2019, 48: 201-205.

[13] Brunner P, Bihl M, Jundt G, Baumhoer D, Hoeller S. BRAF p.V600E mutations are not unique to ameloblastoma and are shared by other odontogenic tumors with ameloblastic morphology. Oral. Oncol., 2015, 51: e77-e78.

[14] Fregnani ER, . BRAF-V600E expression correlates with ameloblastoma aggressiveness. Histopathology, 2017, 70: 473-484.

[15] Pereira NB, . BRAFV600E mutation in the diagnosis of unicystic ameloblastoma. J. Oral. Pathol. Med., 2016, 45: 780-785.

[16] Brown NA, . Activating FGFR2-RAS-BRAF mutations in ameloblastoma. Clin. Cancer Res., 2014, 20: 5517-5526.

[17] Diniz MG, . Assessment of BRAFV600E and SMOF412E mutations in epithelial odontogenic tumours. Tumour Biol., 2015, 36: 5649-5653.

[18] Soltani M, . Genetic study of the BRAF gene reveals new variants and high frequency of the V600E mutation among Iranian ameloblastoma patients. J. Oral. Pathol. Med., 2018, 47: 86-90.

[19] Sweeney RT, . Identification of recurrent SMO and BRAF mutations in ameloblastomas. Nat. Genet., 2014, 46: 722-725.

[20] Yukimori A, . Genetic basis of calcifying cystic odontogenic tumors. PLoS ONE, 2017, 12: e0180224.

[21] do Canto AM, . Immunohistochemical analysis of BRAF V600E mutation in ameloblastomas. Clin. Oral. Investig., 2019, 23: 779-784.

[22] Davanian H, . Ameloblastoma RNA profiling uncovers a distinct non-coding RNA signature. Oncotarget, 2017, 8: 4530-4542.

[23] Amm HM, MacDougall M. Molecular signaling in benign odontogenic neoplasia pathogenesis. Curr. Oral. Health Rep., 2016, 3: 82-92.

[24] Gultekin SE, . The landscape of genetic alterations in ameloblastomas relates to clinical features. Virchows Arch., 2018, 472: 807-814.

[25] Bologna-Molina R, . Detection of MAPK/ERK pathway proteins and KRAS mutations in adenomatoid odontogenic tumors. Oral. Dis., 2019, 25: 481-487.

[26] Heikinheimo K, . The mutational profile of unicystic ameloblastoma. J. Dent. Res., 2019, 98: 54-60.

[27] Zhang R, . The BRAF p.V600E mutation is a common event in ameloblastomas but is absent in odontogenic keratocysts. Oral. Surg. Oral. Med Oral. Pathol. Oral. Radiol., 2020, 129: 229-235.

[28] Xia RH, . Ameloblastoma with mucous cells: a clinicopathological, BRAF mutation, and MAML2 rearrangement study. Oral. Dis., 2020, 26: 805-814.

[29] Agaimy A. The expanding family of SMARCB1(INI1)-deficient neoplasia: implications of phenotypic, biological, and molecular heterogeneity. Adv. Anat. Pathol., 2014, 21: 394-410.

[30] Kondo S, . Discovery of novel molecular characteristics and cellular biological properties in ameloblastoma. Cancer Med., 2020, 9: 2904-2917.

[31] Fumagalli D, . A rapid, sensitive, reproducible and cost-effective method for mutation profiling of colon cancer and metastatic lymph nodes. BMC Cancer, 2010, 10

[32] Perkins G, . Multi-purpose utility of circulating plasma DNA testing in patients with advanced cancers. PLoS ONE, 2012, 7: e47020.

[33] Stemke-Hale K, . Frequency of mutations and polymorphisms in borderline ovarian tumors of known cancer genes. Mod. Pathol., 2013, 26: 544-552.

[34] Kurppa KJ, . High frequency of BRAF V600E mutations in ameloblastoma. J. Pathol., 2014, 232: 492-498.

[35] Duarte-Andrade FF, . The importance of BRAF-V600E mutation to ameloblastoma metabolism. J. Oral. Pathol. Med., 2019, 48: 307-314.

[36] Seki-Soda M, Sano T, Ito K, Yokoo S, Oyama T. An immunohistochemical and genetic study of BRAF(V600E) mutation in Japanese patients with ameloblastoma. Pathol. Int., 2020, 70: 224-230.

[37] Oh KY, . High prevalence of BRAF V600E mutations in Korean patients with ameloblastoma: Clinicopathological significance and correlation with epithelial-mesenchymal transition. J. Oral. Pathol. Med., 2019, 48: 413-420.

[38] Derakhshan S, . High frequency of BRAF V600E mutation in Iranian population ameloblastomas. Med. Oral. Patol. Oral. Cir. Bucal, 2020, 25: e502-e507.

[39] Guan P, . Mutational signatures in mandibular ameloblastoma correlate with smoking. J. Dent. Res., 2019, 98: 652-658.

[40] Kreppel M, Zoller J. Ameloblastoma-clinical, radiological, and therapeutic findings. Oral. Dis., 2018, 24: 63-66.

[41] Oh, K. Y., Cho, S. D., Yoon, H. J., Lee, J. I. & Hong, S. D. Discrepancy between immunohistochemistry and sequencing for BRAF V600E in odontogenic tumours: comparative analysis of two VE1 antibodies. J. Oral Pathol. Med. https://doi.org/10.1111/jop.13108 (2020).

[42] Lapthanasupkul, P., Laosuk, T., Ruangvejvorachai, P., Aittiwarapoj, A. & Kitkumthorn, N. Frequency of BRAF V600E mutation in a group of Thai patients with ameloblastomas. Oral Surg Oral Med Oral Pathol Oral Radiol. https://doi.org/10.1016/j.oooo.2020.06.002 (2020).

[43] Heikinheimo K, Kurppa KJ, Elenius K. Novel targets for the treatment of ameloblastoma. J. Dent. Res., 2015, 94: 237-240.

[44] Gonzalez-Gonzalez R, . Current concepts in ameloblastoma-targeted therapies in B-raf proto-oncogene serine/threonine kinase V600E mutation: systematic review. World J. Clin. Oncol., 2020, 11: 31-42.

[45] Tan S, Pollack JR, Kaplan MJ, Colevas AD, West RB. BRAF inhibitor therapy of primary ameloblastoma. Oral. Surg. Oral. Med. Oral. Pathol. Oral. Radio., 2016, 122: 518-519.

[46] Niu Z, . Study on clinical and biological characteristics of ameloblastic carcinoma. Orphanet J. Rare Dis., 2020, 15

[47] Bologna-Molina R, . Primordial odontogenic tumor: an immunohistochemical profile. Med Oral. Patol. Oral. Cir. Bucal, 2017, 22: e314-e323.

[48] Coura BP, . Assessing pathogenic mutations in dental follicles as an attempt to identify early events in odontogenic tumours tumourigenesis. Arch. Oral. Biol., 2020, 113: 104523.

[49] Cha YH, . Frequent oncogenic BRAF V600E mutation in odontogenic keratocyst. Oral. Oncol., 2017, 74: 62-67.

[50] Franca JA, . Absence of BRAFV600E mutation in odontogenic keratocysts. J. Oral. Pathol. Med., 2018, 47: 186-191.

[51] Jain KS, . Absence of BRAFV600E immunohistochemical expression in sporadic odontogenic keratocyst, syndromic odontogenic keratocyst and orthokeratinized odontogenic cyst. J. Oral. Pathol. Med., 2020, 49: 1061-1067.

[52] Guimarães LM, . The molecular pathology of odontogenic tumors: expanding the spectrum of MAPK pathway driven tumors. Front. Oral. Health, 2021, 2: 740788.

[53] Bello I, . FAK, paxillin, and PI3K in ameloblastoma and adenomatoid odontogenic tumor. Clin. Oral. Investig., 2021, 25: 1559-1567.

[54] Scheper MA, . Expression and alterations of the PTEN / AKT / mTOR pathway in ameloblastomas. Oral. Dis., 2008, 14: 561-568.

[55] Kumamoto H, OOya K. Immunohistochemical detection of phosphorylated Akt, PI3K, and PTEN in ameloblastic tumors. Oral. Dis., 2007, 13: 461-467.

[56] Capper D, . Assessment of BRAF V600E mutation status by immunohistochemistry with a mutation-specific monoclonal antibody. Acta Neuropathol., 2011, 122: 11-19.

[57] Rotellini M, Maggiore G, Trovati M, Saraceno MS, Franchi A. Metastasizing maxillary ameloblastoma: report of a case with molecular characterization. J. Oral. Maxillofac. Res., 2016, 7: e5.

[58] Chang TH, . LGR5(+) epithelial tumor stem-like cells generate a 3D-organoid model for ameloblastoma. Cell Death Dis., 2020, 11

[59] Kaye FJ, Ivey AM, Drane WE, Mendenhall WM, Allan RW. Clinical and radiographic response with combined BRAF-targeted therapy in stage 4 ameloblastoma. J. Natl Cancer Inst., 2015, 107: 378.

[60] Faden, D. L. & Algazi, A. Durable treatment of ameloblastoma with single agent BRAFi Re: clinical and radiographic response with combined BRAF-targeted therapy in stage 4 ameloblastoma. J. Natl Cancer Inst. 109, https://doi.org/10.1093/jnci/djw190 (2017).

[61] Fernandes GS, Girardi DM, Bernardes JPG, Fonseca FP, Fregnani ER. Clinical benefit and radiological response with BRAF inhibitor in a patient with recurrent ameloblastoma harboring V600E mutation. BMC Cancer, 2018, 18

[62] Broudic-Guibert M, . Persistent response to vemurafenib in metastatic ameloblastoma with BRAF mutation: a case report. J. Med Case Rep., 2019, 13

[63] Brunet M, Khalifa E, Italiano A. Enabling precision medicine for rare head and neck tumors: the example of BRAF/MEK targeting in patients with metastatic ameloblastoma. Front. Oncol., 2019, 9: 1204.