BRAFV600E mutation in the diagnosis of unicystic ameloblastoma.

Unicystic ameloblastoma, an odontogenic neoplasm, presents clinical and radiographic similarities with dentigerous and radicular cysts, non-neoplastic lesions. It is not always possible to reach a final diagnosis with the incisional biopsy, leading to inappropriate treatment. The BRAFV600E activating mutation has been reported in a high proportion of ameloblastomas. The purpose of the study was to assess the utility of the detection of the BRAFV600E mutation in the differential diagnosis of unicystic ameloblastoma with dentigerous and radicular cysts. Twenty-six archival samples were included, comprising eight unicystic ameloblastomas (UAs), nine dentigerous and nine radicular cysts. The mutation was assessed in all samples by anti-BRAFV600E (clone VE1) immunohistochemistry (IHC) and by TaqMan mutation detection qPCR assay. Sanger sequencing was further carried out when samples showed conflicting results in the IHC and qPCR. Although all UAs (8/8) showed positive uniform BRAFV600E staining along the epithelial lining length, the mutation was not confirmed by qPCR and Sanger sequencing in three samples. Positive staining for the BRAFV600E protein was observed in one dentigerous cyst, but it was not confirmed by the molecular methods. Furthermore, 2/9 dentigerous cysts and 2/9 radicular cysts showed non-specific immunostaining of the epithelium or plasma cells. None of the dentigerous or radicular cysts cases presented the BRAFV600E mutation in the qPCR assay. The BRAFV600E antibody (clone VE1) IHC may show non-specific staining, but molecular assays may be useful for the diagnosis of unicystic ameloblastoma, in conjunction with clinical, radiological and histopathological features.

Pereira NB ，Pereira KM ，Coura BP ，Diniz MG ，de Castro WH ，Gomes CC ，Gomez RS ... -  《-》

Assessment of BRAFV600E and SMOF412E mutations in epithelial odontogenic tumours.

Diniz MG ，Gomes CC ，Guimarães BV ，Castro WH ，Lacerda JC ，Cardoso SV ，de Faria PR ，Dias FL ，Eisenberg AL ，Loyola AM ，Gomez RS ... -  《-》

Diagnostic dilemma of unicystic ameloblastoma: novel parameters to differentiate unicystic ameloblastoma from common odontogenic cysts.

Diagnostic criteria that have been specified for unicystic ameloblastomas (UAs) are not always helpful to differentiate these cystic tumors from common odontogenic cysts. The aim of this study therefore was to identify additional histopathological features (other than the features considered for the diagnosis of UA at present) that would be helpful to differentiate UA from odontogenic cysts. One hundred histopathologically confirmed unicystic ameloblastomas and 20 cases each of radicular, inflamed dentigerous and non-inflamed dentigerous cysts were selected. Histopathological features of the UAs that are not used as diagnostic criteria at present were identified. Hyperplastic arcading epithelial proliferations with stellate-reticulum-like and vacuolated cells were always seen associated with inflammation in odontogenic cysts, while in UA plexiform-like areas were also seen without inflammation (P < 0.001). In addition, a spiky rete pattern was observed in non-inflamed UA while this pattern was observed only in inflamed odontogenic cysts. Furthermore, spiky retes together with subepithelial hyalinization were usually observed in UAs while only subepithelial hyalinization was observed in non-inflamed dentigerous cysts. Combinations of histopathological features were identified to differentiate non-inflamed UA from common odontogenic cysts. However, presence of inflammatory changes in UA precludes the use of features identified in the present study for diagnostic purposes.

Gunawardhana KS ，Jayasooriya PR ，Tilakaratne WM 《-》

The diagnostic utility of BRAF VE1 mutation-specific immunohistochemistry in ameloblastoma.

Ameloblastoma is a benign, locally aggressive odontogenic neoplasm with variable solid and cystic morphology. On account of its histologic variety, diagnostically challenging cases can bear resemblance to odontogenic keratocyst/keratocystic odontogenic tumor (KCOT) or dentigerous cyst (DC). BRAFV600E mutation has been reported to be specific for and frequent in ameloblastoma, and this study evaluated the usefulness of immunohistochemistry (IHC) using the BRAF VE1 mutant-specific antibody as a diagnostic adjunct in this setting. We investigated 46 ameloblastomas, 30 KCOTs, and 30 DCs. BRAF VE1 IHC was performed on all cases and allele-specific polymerase chain reaction (AS-PCR) for BRAFV600E mutation was performed on 30 ameloblastomas and any IHC-positive KCOT/DC. BRAF VE1 IHC was positive in 31/37 (83.8%) mandibular ameloblastomas but not in any maxillary ameloblastomas (0/9), KCOT (0/30), or DC (0/30). Equivocal staining was seen in 1/37 (3.3%) mandibular ameloblastomas. Of the 30 ameloblastomas subjected to AS-PCR, BRAFV600E mutation was identified in 19/23 (82.6%) mandibular ameloblastomas and 0/7 (0.0%) maxillary ameloblastomas. BRAFV600E mutant ameloblastomas were positive by IHC in 18/19 (94.7%) cases and equivocal in 1/19 (5.3%) cases. All 11 (100.0%) BRAF-wild type ameloblastomas were negative by IHC. BRAF VE1 is an excellent tool for the diagnosis of mandibular ameloblastoma but of limited utility in the maxilla, where it less commonly occurs and where BRAFV600E mutation is considerably less frequent.

Mendez LD ，Wolsefer NS ，Asa SL ，Wasman J ，Yoest JM ，Stojanov IJ ... -  《-》

BRAF p.V600E status in epithelial areas of ameloblastoma with different histological aspects: Implications to the clinical practice.

BRAF p.V600E is reported in up to 80% of ameloblastomas. Despite the high frequency, the presence of this mutation in different histopathological areas of the tumour has not been investigated. This information has an important role in the use of BRAF p.V600E assessment as an auxiliary tool in the differential diagnosis between unicystic ameloblastoma and other odontogenic cystic lesions, especially when only incisional biopsies are available. Therefore, the purpose of the present study was to investigate BRAF p.V600E heterogeneity in unicystic ameloblastoma. Five cases of ameloblastoma and two dentigerous cysts were analysed. The regions exhibiting different microscopic characteristics were selected from each ameloblastoma case and manually dissected. TaqMan allele-specific qPCR or Sanger sequencing was performed to determine BRAF p.V600E status. We screened the mutation in a small cohort of UA and no molecular heterogeneity was found. Four cases of ameloblastoma (80%) exhibited BRAF p.V600E in all different areas evaluated. One case did not harbour the mutation in any microscopic region analysed. The BRAF mutation was absent in the dentigerous cysts. Ameloblastomas appear to exhibit a homogeneous profile regarding the BRAF p.V600E no matter what histological feature is observed under light microscopy, suggesting that this molecular test may contribute to establish the correct diagnosis in cases microscopically resembling other odontogenic lesions.

Sant'Ana MSP ，Dos Santos Costa SF ，da Silva MP ，Martins-Chaves RR ，Pereira TDSF ，de Oliveira EM ，Martínez Pedraza R ，de Castro WH ，Gomes CC ，Gomez RS ，Fonseca FP ... -  《-》