<i>BRAF</i> Mutation Analysis of Fine-Needle Aspiration Biopsies of Papillary Thyroid Carcinoma: Impact on Diagnosis and Prognosis

Abstract

Objective: The BRAF V600E mutation has been associated with aggressive disease in papillary thyroid carcinoma (PTC). Molecular testing has been proposed as a useful adjunct to cytology in the diagnosis of malignancy and for tailoring clinical management. The aims of our study were to evaluate the BRAF mutational status using archived fine-needle aspiration biopsy (FNAB) material from patients with long-term follow-up and to correlate it with the original cytology diagnosis, clinicopathological stage at surgery, and prognosis. Study Design: FNAB material from 52 cases of PTC, with a mean follow-up of 8.4 years, was used in this study. DNA was extracted from archival cytology slides. Mutation analysis was performed by standard sequencing and locked nucleic acid-PCR/sequencing. Results: The BRAF V600E mutation was present in 46% of cases, but it was absent in all FNABs diagnosed originally as atypical and in 14 of 17 suspicious cases. Recurrence was significantly more frequent (p = 0.006) in cases with BRAF mutations and 54% of these cases presented with stage 2 or higher. Conclusion: The BRAF V600E mutation is associated with a higher pathological stage at surgery and a higher rate of recurrence. BRAF mutation analysis did not provide a significant increase in the accuracy of thyroid FNABs diagnosed as suspicious or atypical in our institution.

Next-generation sequencing-based multi-gene mutation profiling of solid tumors using fine needle aspiration samples: promises and challenges for routine clinical diagnostics

“…[17][18][19][20]23 Detection of BRAF mutation in such cases strongly favors a diagnosis of papillary thyroid carcinoma. 12,[24][25][26][27] Second, the presence of mutation can help stratify patients into less aggressive or more aggressive groups. 12,25,28,29 Third, a cytological specimen may be the sole diagnostic material available for a patient's tumor.…”

“…12,[24][25][26][27] Second, the presence of mutation can help stratify patients into less aggressive or more aggressive groups. 12,25,28,29 Third, a cytological specimen may be the sole diagnostic material available for a patient's tumor. For example, only a third of the lung carcinomas are resectable at the time of initial presentation.…”

“…For example, only a third of the lung carcinomas are resectable at the time of initial presentation. 12 If the lung mass is large, locally advanced or metastatic, or is located in an inaccessible site, excisional biopsy or resection cannot be performed. 1,6 We also believe that FNA samples may be more representative of the tumor due to sampling of multiple areas of tumor by this technique compared with the unidirectional core needle biopsy.…”

“…1 Additionally, use of COLD-PCR/direct sequencing for EGFR and KRAS detection in lung aspirates and locked nucleic acid-based PCR/sequencing assay in thyroid aspirates has improved the sensitivity of mutation detection. 10,12 Massively parallel or next-generation sequencing (NGS) technology is increasingly being used for mutational analysis of tumors for both clinical and research applications. The advent of affordable bench top NGS platforms such as Ion Personal Genome Machine (PGM) has facilitated multi-gene mutational profiling using only nanograms (ng) of DNA.…”

Assessment of cellularity, genomic DNA yields, and technical platforms for BRAF mutational testing in thyroid fine‐needle aspirate samples

“…12 BRAF mutations are fairly specific for PTC, not being found in benign thyroid lesions or other types of thyroid cancers. 13 In addition, BRAF mutations have consistently been associated with more aggressive BRAF mutational analysis has traditionally been performed on formalin-fixed paraffin-embedded surgical tissue. Recently, several studies have been published on the feasibility of successfully testing for BRAF mutations using cytologic materials.…”