BRAFV600E Mutation In Papillary Thyroid Carcinoma Research Articles

Application of Ultrasound Shear Wave Elastography Combined With BRAFV600E Mutation for the Diagnosis of Papillary Thyroid Carcinoma.

This prospective study aimed to investigate the combination of shear wave elastography (SWE) and BRAFV600E mutation testing for the diagnosis of papillary thyroid carcinoma (PTC). One hundred thyroid nodules with an American College of Radiology thyroid imaging reporting and data system classification of 4 or 5 were subjected to SWE measurement, BRAFV600E genotyping, fine-needle aspiration, and surgery. Nonparametric statistical tests were used to compare the differences in the elastic parameters and BRAF genotypes between benign and malignant thyroid nodules of PTC, and receiver operating characteristic curve analysis was conducted to compare the diagnostic efficacy. In addition, the correlations between elastic parameters and BRAFV600E mutation in PTC were analyzed using binary logistic regression. The SWE, BRAFV600E, and their combination exhibited sensitivities of 72.9%, 81.3%, and 85.4%, respectively, and specificities of 66.7%, 90.3%, and 86.5%, respectively, in the diagnosis of PTC ( P < 0.05). The SWE, BRAFV600E, and their combination exhibited sensitivities of 50.0%, 24.1% and 56.3%, respectively, and specificities of 89.7%, 87.5% and 82.8%, respectively, in the diagnosis of central cervical lymph node metastasis ( P < 0.05). The combined use of SWE and BRAFV600E detection had the largest area under the curve, indicating that this combination is more effective in diagnosing PTC and lymph node metastasis in the central region than either method alone. Furthermore, Emax was positively associated with the BRAFV600E genotype. In conclusion, the combination of SWE and BRAFV600E genotype detection can improve the diagnostic efficacy for PTC. Emax can predict the BRAFV600E mutation status.

Iodine nutritional status is not a direct factor in the prevalence of the BRAFV600E mutation in papillary thyroid cancer.

Papillary thyroid carcinoma (PTC) accounts for approximately 85%-90% of all thyroid cancers. Of the iodine-metabolizing genes, BRAFV600E is a highly specific target for PTC and may have a reciprocal causative relationship with iodide-metabolizing genes. In this study, we performed a data analysis of selected quantitative studies to determine the relationship between iodine nutritional status and the prevalence of the BRAF600E mutation in patients with PTC. Five studies were selected for meta-analysis based on the selection criteria. A total of 2,068 patients were divided into three groups: low (urinary iodine concentration [UIC] < 100 µg/L), adequate (UIC 100-200 µg/L), and high (UIC ≥ 200 µg/L). The results were obtained using RevMan software, and the pooled odds ratios (ORs) were calculated using Mantel-Haenszel statistics with a 95% confidence interval (CI). The OR for the prevalence of the BRAFV600E mutation between the high and adequate groups was 1.25 (95% CI 0.64-2.43, p = 0.51), and the OR between the low and adequate groups was 0.98 (95% CI 0.42-2.31, p = 0.96). The BRAFV600E mutation risk did not change significantly at different levels of iodine nutrition (p = 0.33) in statistical analyses. We conducted preliminary research on dietary iodine intake and the BRAFV600E mutation in PTC. The results suggested that abnormal iodine intake might not directly influence the prevalence of the BRAFV600E mutation in these patients. Further research into the associations between dietary iodine intake and the BRAFV600E mutation in PTC, including the underlying mechanisms, is required.

MRI-Based Texture Analysis for Preoperative Prediction of BRAF V600E Mutation in Papillary Thyroid Carcinoma

BRAF V600E mutation can compensate for the low detection rate by fine-needle aspiration (FNA) and is related to aggressiveness and lymph node metastasis. This study aimed to investigate the relationship between texture analysis features based on magnetic resonance imaging (MRI) and mutations. Retrospective analysis was performed on patients with postoperative pathology confirmed papillary thyroid carcinoma (PTC) from 2017 to 2021. One thousand one hundred and thirty-two texture features were extracted from T2-weighted imaging (T2WI) and contrast-enhanced T1-weighted imaging (CE-T1WI) separately by outlining the tumor volume of interest (VOI). Univariate, minimum redundancy maximum relevance (mRMR), and multivariate analyses were used for feature selection to construct 3 models (T2WI, CE-T1WI, and combined model) to predict mutation. The reproducibility between observers was evaluated by intraclass correlation coefficient (ICC). Receiver operating characteristic (ROC) analysis was used to assess the performance of models. The diagnostic performance of the optimal cut-off value of models were calculated and validated by 10-fold cross-validation. A total of 80 PTCs (22 BRAF V600E wild-type and 58 BRAF V600E mutant) were included in our study. Good interobserver agreement was found on texture features we selected (all ICCs >0.75). The area under the ROC curves (AUCs) for the T2WI model, CE-T1WI model, and combined model were 0.83 (95% CI: 0.75-0.91), 0.83 (95% CI: 0.73-0.90), and 0.88 (95% CI: 0.81-0.94), respectively. The accuracy, sensitivity, specificity, PPV, and NPV were 0.776, 0.679, 0.905, 0.905, and 0.679 for the T2WI model at a cut-off value of 0.674; 0.755, 0.750, 0.762, 0.808, and 0.696 for the CE-T1WI model at a cut-off value of 0.573; 0.816, 0.893, 0.714, 0.806, and 0.833 for the combined model at a cut-off value of 0.420. MRI-based texture analysis could be a potential method for predicting BRAF V600E mutation in PTC preoperatively.

BRAFV600E Mutation in Papillary Carcinoma Thyroid

Background: Thyroid malignancies comprise 1 % of total human malignancies, Papillary Thyroid Carcinoma (PTC) is the most common malignant tumour of thyroid cancers. It is estimated that 80 % of thyroid malignancies are comprised of papillary carcinoma thyroid, which shows increased incidence in women. The most important etiology of PTC is mutation of RET/RAS/BRAF which activates MAPK signaling pathway. The BRAF V600E activating point mutation appears to be highly specific for PTC.&#x0D; Methods: It is cross sectional study of 56 cases studied in the Government Medical college, Thiruvananthapuram, Kerala. Consecutive cases of histo-pathologically proven cases were selected and Immunohistochemical (IHC) studies for BRAFV600E were conducted, and results were recorded.&#x0D; Result: Our study showed female preponderance and equal distribution of cases above and below 45 years of age. Size of the primary tumour was more than 1cm for majority of cases. In our study out of 56 cases studied 45 patients showed BRAFV600E expression comprising of 80% positivity. A 44% of the total cases studied showed lymph node metastasis.&#x0D; Conclusion: We conclude in our study that BRAFV600E mutation is most common mutation in primary PTC patients. BRAFV600E expression most seen in elderly patients and shows strong association with lymph node metastasis. IHC assay is a reliable method to detect BRAFV600E mutation in PTC and can be used as alternative to molecular methods on routine practice

BRAFV600E Mutation-Responsive miRNA-222-3p Promotes Metastasis of Papillary Thyroid Cancer Cells via Snail-Induced EMT.

BRAF mutation accounts for 50% of the PTC (papillary thyroid carcinoma) and is closely associated with high-risk clinicopathological characteristics. Increasing evidence implied that dysregulation of miRNA participated in carcinogenesis and progression of cancer. Clinical data showed the significant up-regulation of miR-222-3p in PTC; however, the role of miR-222-3p and possible relationship with BRAF mutation remained unclear. Here, we identified significant up-regulation of miR-222-3p in PTC tissues harboring BRAFV600E mutation compared with BRAF wild type (BRAFWT ) from collected PTC clinical samples. External validation performed with The Cancer Genome Atlas (TCGA) databases was consistent with the above result. Exogenous expression of BRAFV600E oncoprotein increased the expression of miR-222-3p in B-CPAP and TPC-1 cells. The treatment of BRAFV600E and MEK inhibitor, PLX4720 and PD0325901, decreased the expression of miR-222-3p in B-CPAP but not in TPC-1. Inhibition of miR-222-3p significantly suppressed the migration of B-CPAP and induced a mesenchymal-epithelial transition (MET) phenotype via the Snail transcription factor. Immunohistochemistry (IHC) analysis demonstrated the up-regulation of Snail correlated with lymph node metastasis and BRAFV600E mutation in PTC. Besides, in situ hybridization (ISH) and IHC analysis of PTC clinical samples confirmed the correlation between the expression of miR-222-3p and Snail. These results showed miR-222-3p conduced more aggressive clinical manifestation of PTC by promoting Snail-induced EMT.

An affordable immunohistochemical approach to estimate the prevalence of BRAFV600E in large cohort studies-establishing the baseline rate of BRAF mutation in an institutional series of papillary thyroid carcinoma from Thailand.

Papillary thyroid carcinoma (PTC) accounts for the majority of diagnoses of thyroid carcinoma. BRAFV600E mutation is the most common genetic alteration in PTC, which has diagnostic and prognostic significance. The rate of BRAFV600E mutation in PTC from Thailand has not been reported. Our purpose was to estimate the prevalence of BRAF mutation in a large institutional series using an affordable approach, which combined mutation-specific immunohistochemistry (IHC) with VE1 antibody and tissue microarray (TMA). A total of 476 PTC cases plotted on TMA were employed for determining the mutation status in this study. The cancer tissue of initial 100 cases (pilot study) were analyzed for BRAFV600E mutation by using both direct sequencing and VE1 immunostaining. For the subsequent PTC cases, VE1 IHC was used as an alternative to direct sequencing for the detection of mutation. Univariate and multivariate analyses were done to determine the association of clinicopathological variables with BRAFV600E mutation. In the pilot study, VE1 IHC showed excellent analytical performance (κ=0.884) for detecting BRAFV600E mutation in PTC TMA as compared to direct sequencing. The prevalence of BRAFV600E in the whole cohort was 60.9% by using VE1 IHC. The mutation was commonly seen in tall cell (92.9%) and classic (70.2%) variants of PTC. Multivariate analysis (P<0.05) showed association of BRAFV600E with histological type of tumor, extrathyroidal extension, and absence of Hashimoto's thyroiditis. In conclusion, BRAFV600E mutation was detected in 60.9% of Thai PTC and it was associated with several aggressive clinicopathological variables of thyroid cancer. VE1 IHC proved as a reliable method able to replace direct sequencing for detection of the mutation. A combination of mutation-specific IHC and TMA allows conducting large cohort studies more labor-saving and cost-efficiently.

Inhibition of BRAF Sensitizes Thyroid Carcinoma to Immunotherapy by Enhancing tsMHCII-mediated Immune Recognition.

Multiple mechanisms play roles in restricting the ability of T-cells to recognize and eliminate tumor cells. To identify immune escape mechanisms involved in papillary thyroid carcinoma (PTC) to optimize immunotherapy. iTRAQ analysis was conducted to identify proteins differentially expressed in PTC samples with or without BRAFV600E mutation. Molecular mechanisms regulating tumor cell evasion were investigated by in vitro modulations of BRAF/MAPK and related pathways. The pathological significance of identified tumor-specific major histocompatibility complex class II (tsMHCII) molecules in mediating tumor cell immune escape and targeted immune therapy was further evaluated in a transgenic mouse model of spontaneous thyroid cancer. Proteomic analysis showed that tsMHCII level was significantly lower in BRAFV600E-associated PTCs and negatively correlated with BRAF mutation status. Constitutive activation of BRAF decreased tsMHCII surface expression on tumor cells, which inhibited activation of CD4+ T-cells and led to immune escape. Pathway analysis indicated that the transforming growth factor (TGF)-β1/SMAD3-mediated repression of tsMHCII, which could be reversed by BRAF inhibition (BRAFi). Targeting this pathway with a combined therapy of BRAF inhibitor PLX4032 and anti-PD-1 antibody efficiently blocked tumor growth by increasing CD4+ T-cell infiltration in a transgenic PTC mouse model. Our results suggest that BRAFV600E mutation in PTC impairs the expression of tsMHCII through the TGF-β1/SMAD3 pathway to enhance immune escape. Combined treatment with PLX4032 and anti-PD-1 antibody promotes recognition and elimination of PTC by the immune system in a pre-clinical mouse model, and therefore offers an effective therapeutic strategy for patients with advanced PTC.

Elevated BANCR expression levels have different effects on papillary thyroid carcinoma progression depending on the presence of the BRAFV600E mutation

IntroductionThe role of BRAF-activated non-protein coding RNA (BANCR) in papillary thyroid carcinoma (PTC) is controversial, its clinical significance is unclear and no study has correlated the presence of the BRAFV600E mutation in PTC with BANCR expression. MethodsBANCR levels in PTC and matched nonmalignant thyroid epithelial tissues from 85 patients were determined using quantitative RT-PCR. BRAFV600E was detected by mutant allele-specific PCR amplification. The results were correlated with clinicopathological characteristics of the patients. ResultsThe presence of BRAFV600E associates with lower relative BANCR expression (RBE) in PTC (p = 0.008). RBE is down-regulated in BRAFV600E positive PTC, while it is unchanged or up-regulated in BRAFV600E negative PTC compared to the levels in paired nonmalignant tissue (p = 0.001). At the cut-off of 31.3%, sensitivity of fold change of BANCR for the presence of BRAFV600E is 68.0% and specificity is 67.2%. In BRAFV600E positive PTC up-regulated BANCR predicts lymph node metastasis (p = 0.001), while in BRAFV600E negative PTCs high RBE predicts thyroid capsule invasion (p = 0.028). ConclusionsDepending on the presence of BRAFV600E, elevated BANCR levels demonstrated different effects on lymphatic spreading and local PTC invasion. Therefore, BANCR could be a useful prognostic biomarker in risk stratification of PTC patients.

VE1 Immunohistochemistry Improves the Limit of Genotyping for Detecting BRAFV600E Mutation in Papillary Thyroid Cancer.

Detection of BRAFV600E is useful for making diagnosis and risk stratification of papillary thyroid carcinoma (PTC). Molecular testing, however, is not always available for routine clinical use. To assess the clinical utility and reliability of VE1 immunohistochemistry (IHC) for detecting BRAFV600E mutation in PTC, VE1 IHC was performed on the tissue microarrays of 514 patients with PTC and was compared with Sanger sequencing results. Of 514 PTC cases, 433 (84.2%) were positive for VE1 expression. Among 6 discordant cases between VE1 IHC and Sanger sequencing, 3 initial VE1-false negative cases turned out to be true false negative on repeat testing, and 3 VE1-false positive cases showed BRAFV600E mutation using digital PCR analysis. PTCs with low variant allele fraction were positive for VE1 IHC but were not detected using sequencing. VE1 IHC showed 99.3% sensitivity, 100% specificity, 100% positive predictive value, and 96.4% negative predictive value. The BRAFV600E mutation was significantly associated with older age, multifocality, extrathyroidal extension, lymph node metastasis, and advanced tumor stage. In conclusion, VE1 IHC is a reliable method for detecting BRAFV600E mutation in PTC specimens.

Analysis of clinicopathological characteristics of papillary thyroid carcinoma and BRAF V600E gene mutation

Objective: To study the correlation between the BRAF V600E mutation and the clinicopathological features of papillary thyroid carcinoma(PTC), and to explore the application value of BRAF V600E mutation in PTC. Method: The BRAF V600E gene mutation was detected in 108 PTC cases and 30 control cases by qPCR. The results were statistically analyzed by using SPSS 21.0 software. Result: The BRAF V600E mutation was absent in the control group. The BRAF V600E mutation rate was 72.20% in 108 PTC cases. Compared with patients with wild type,the mean age of patients with the mutant BRAF V600E was significantly older(P<0.05). BRAF V600E mutations were associated with PTC tissue subtypes and central area lymph node metastasis(P<0.05). BRAF V600E mutations were not correlated with tumor diameter, patient gender, multifocality, tumor site, concomitant disease, lymph node metastasis in side region of neck, capsule invasion, and TNM staging(P>0.05). Conclusion: The value of BRAF V600E mutation in the prognosis of PTC remains to be studied. BRAF V600E mutation is of value for pathological diagnosis of PTC.

CTLA-4 and PD-1 Ligand Gene Expression in Epithelial Thyroid Cancers.

The dysregulation of PD-1 ligands (PD-L1 and PD-L2) and CTLA-4 ligands (CD80 and CD86) represents a tumor strategy to escape the immune surveillance. Here, the expression of PD-L1, PD-L2, CD80, and CD86 was evaluated at the mRNA level in 94 patients affected by papillary thyroid carcinoma (PTC) and 11 patients affected by anaplastic thyroid carcinoma (ATC). Variations in the mRNAs in PTC patients were then correlated with clinicopathological features. The expression of all genes was deregulated in PTC and ATC tissues compared to normal tissues. In particular, the downregulation of CD80 was observed above all in ATC. In addition, the increased expression of CD80 associated with longer disease-free survival in PTC. Higher expression of PD-L1 associated with the classical histological variant and with the presence of BRAFV600E mutation in PTC. The increased PD-L2 expression correlated with BRAFV600E mutation and lymph node metastasis, while its lower expression correlated with the follicular PTC variant. The latter was also associated with the CD80 downregulation, which was also related to the absence of lymph node metastasis. In conclusion, we documented the overall dysregulation of PD-1 and CTLA-4 ligands in PTC and ATC tissues and a possible prognostic value for CD80 gene expression in PTC.

Investigation of BRAF V600E detection approaches in papillary thyroid carcinoma

ObjectiveThe detection of BRAF V600E mutation in papillary thyroid carcinoma (PTC) may be helpful to offer diagnostic confirmation. Additionally, such detection may provide a targeted therapeutic approach for the radioactive iodine resistant patients to predict adverse outcomes. To compare the results of immunohistochemistry (IHC) method using the anti-BRAF V600E (VE1) antibody with the Quantitative real-time polymerase chain reaction (qPCR) approach in examining BRAF V600E mutation in PTC, we investigated the sensitivity and specificity of BRAF V600E (clone VE1) mouse monoclonal antibody in detecting the BRAF V600E mutation and correlated BRAF V600E mutation with clinicopathologic features in PTC. MethodsIHC and qPCR were performed in 40 cases of paraffin-embedded PTCs tissues. The association between BRAFV600E mutation and clinicopathologic features of PTC was assessed with the χ2 test. ResultsThe concordance rate between IHC and qPCR analyses was 95% (38/40). The BRAF V600E (VE1) antibody has a sensitivity of 100% (34/34) and specificity of 66.67% (4/6) for detecting the mutation. Our study showed that there was no significant association of BRAF V600E mutation with the gender, age, tumor size and lymph node metastasis in PTCs. ConclusionWe may draw the conclusion that detection of BRAF V600E mutation by immunohistochemistry is highly sensitive and specific. Immunohistochemical detection of the mutated BRAF V600E protein in PTC may facilitate mutational analysis in the clinical setting.

V-raf murine sarcoma viral oncogene homolog B1 V600E mutation and urinary iodine concentration in papillary thyroid carcinoma

Objective To investigate the V-raf murine sarcoma viral oncogene homolog B1 (BRAF) V600E mutation and urinary iodine concentration in papillary thyroid carcinoma (PTC) and the relationship with clinicopathological features. Methods Polymerase chain reaction (PCR) and gene sequencing were performed to detect BRAF V600E mutation in 110 cases of PTC, 47 cases of nodular goiter and 26 of thyroid adenoma thyroid tissues. Simultaneously, the urinary iodine concentration was determined. The relationship between BRAF V600E mutation and urinary iodine concentration with clinicopathological features was studied. Results In 183 cases, BRAF V600E mutation was detected in 54 out of 110 PTC cases with a detection rate of 49% (χ2=50.838, P=0.000). BRAF V600E mutation was negative in nodular goiter and thyroid adenoma. BRAF V600E mutation in PTC was positively correlated with extra-thymidal extension (χ2=6.031, P=0.014), but not with gender (P=0.363), age (P=0.360), tumor number (P=0.593), tumor size (P=0.497), lymph nodal metastasis (P=0.554) and pTNM staging (P=0.785). The excessive iodine intake rate in PTC patients was 79%, obviously higher than that in nodular goiter (66%) and thyroid adenoma (58%) with significant difference (χ2=6.288, P=0.043). The urinary iodine concentration in PTC was negatively correlated with gender (P=0.187), age (P=0.138), tumor number (P=0.571), tumor size (P=0.454), extra-thymidal extension (P=0.345), lymph nodal metastasis (P=0.248) and pTNM staging (P=0.792). The BRAF V600E mutation was not associated with the concentration of iodide in the PTC (χ2=0.019, P=0.891). Conclusion BRAF V600E mutation was a common molecular event and may increase the ability of invasion in PTC. BRAF V600E mutation was a specificity marker in the diagnosis of PTC. The urinary iodine concentration in PTC was significantly higher than that in the benign thyroid nodule diseases. Key words: V-raf murine sarcoma viral oncogene homolog B1 gene; Mutation; Urinary iodine concentration; Papillary thyroid carcinoma

MUC1 expression in papillary thyroid carcinoma is associated with BRAF mutation and lymph node metastasis; the latter is the most important risk factor of relapse.

The incidence of papillary thyroid carcinoma (PTC) has increased over the past 30 years in Western countries. PTC is usually associated with a good prognosis, but there is a wide range of aggressiveness, and some patients develop distant metastasis and/or resistance to standard treatment. Early identification of these high-risk tumors is a current challenge for appropriate patient management. MUC1 expression has been studied previously in thyroid cancer, but its prognostic value remains controversial. Here, we correlated MUC1 expression in PTC with clinical and pathological features and with the presence of the BRAF(V600E) mutation. We performed a clinical and morphological analysis of 190 thyroid tumors (95 PTCs and 95 adenomas). MUC1 immunohistochemistry was carried out on a tissue microarray using different antibodies. The presence of the BRAF(V600E) mutation was investigated by pyrosequencing. MUC1 mRNA levels were assessed by quantitative reverse transcription polymerase chain reaction on a subset of PTC. MUC1 expression was observed in 49% of PTCs and was found to correlate with the presence of papillary architecture, a stromal lymphoid infiltrate, aggressive histological subtypes, extrathyroidal extension, lymph node metastasis, nuclear pseudoinclusions, lymphovascular invasion, and the presence of the BRAF(V600E) mutation (p<0.0001). MUC1 was abundant in nuclear pseudoinclusions. Multivariate analysis showed a strong association of MUC1 expression with the presence of the BRAF(V600E) mutation and lymph node metastasis (p<0.0001). Lymph node metastasis was the most important risk factor of relapse. Our study shows an association between MUC1 expression and the presence of the BRAF(V600E) mutation in PTC. Analysis of MUC1 expression could improve the risk stratification of PTCs.

Time to re-consider the meaning of BRAF V600E mutation in papillary thyroid carcinoma.

The BRAF V600E mutation, resulting from the BRAFT1799A transversion, is the most common genetic mutation in papillary thyroid carcinoma (PTC), with a mean frequency close to 50% among all cases. A large number of studies in the past decade have tried to dissect the relevance and the function of the V600E mutation in controlling oncogenesis and progression of thyroid cancer. However, several works published in the latest years have provided new evidence, in partial conflict with the previous knowledge, suggesting the need of reconsidering the meaning of the BRAF V600E mutation in PTC. In this work, we attempt to discuss some of the most recent molecular, preclinical and clinical evidence to construct a more exhaustive model of function for the BRAF V600E in development, progression and therapeutic approach of thyroid cancer.

Diagnostic Value of Immunohistochemistry for the Detection of the BRAFV600E Mutation in Papillary Thyroid Carcinoma: Comparative Analysis with Three DNA-Based Assays

The aim of this study was to compare the detection of BRAF(V600E) by immunohistochemistry (IHC) using a mutation-specific antibody with molecular biology methods for evaluation of papillary thyroid carcinoma (PTC) patients. This study concerned 198 consecutive conventional PTC patients, of which the majority were women (133/198; 67%), with a mean age of 56 years (range 19-79 years). BRAF mutation analysis was performed using DNA-based (direct sequencing, pyrosequencing, and SNaPshot) and IHC (VE1 antibody) methods. The sensitivity and specificity of IHC for BRAF(V600E) was compared with the molecular biology data. A BRAF mutational result was obtained in 194 cases. A BRAF(V600E) mutation was detected in 153/194 (79%) cases of PTC when using at least one molecular method, and in 151/194 (78%) cases with IHC. No false positive results were noted using IHC to detect the BRAF(V600E) mutation. Besides this mutation, other rare BRAF mutations (BRAF(V600K) and BRAF(K601E)), used as negative controls, were consistently negative with IHC. The sensitivity and specificity of IHC for the detection of this mutation were 98.7% and 100% respectively. The IHC test demonstrated excellent performance at a level equivalent to two DNA-based counterparts (pyrosequencing and SNaPshot). Failure to achieve a result was more frequent with the direct sequencing method than with the three other methods. IHC using the VE1 antibody is a specific and sensitive method for the detection of the BRAF(V600E) mutation in PTC. IHC may be an alternative to molecular biology approaches for the routine detection of this mutation in PTC patients.

Lack of Association ofBRAFMutation With Negative Prognostic Indicators in Papillary Thyroid Carcinoma

Papillary thyroid carcinoma (PTC) is the most common endocrine neoplasm. B-type raf kinase (BRAF) V600E mutation has been proposed as a negative prognostic indicator in PTC, and patients harboring it should receive more aggressive initial therapy. To assess the significance of BRAF V600E mutation in PTC in the largest US sample to date. We identified patients from our institution's pathology archives diagnosed as having PTC and meeting criteria for BRAF mutation testing. Medical records were analyzed for BRAF status (positive or negative) and a list of standardized clinicopathologic features. A total of 429 patients with PTC at an academic medical center. Clinicopathologic features in patients with PTC with and without BRAF mutation. Of 429 cases with PTC, 314 (73.2%) were positive for the BRAF mutation and 115 (26.8%) tested negative. BRAF mutation was significantly associated with tumor margin positivity (P = .03) and lymph node metastasis (P = .002) on univariate analysis but not on multivariate study. BRAF mutation was a predictor of male sex (odds ratio [OR], 3.2; 95% CI, 1.4-7.2), total thyroidectomy (OR, 2.6; 95% CI, 1.1-6.2), and a negative predictor of follicular variant PTC (OR, 0.1; 95% CI, 0.1-0.4). There was no significant association between BRAF positivity and tumor multicentricity, lymphovascular invasion, extranodal extension, central neck involvement, advanced stage (stage III or IV), and distant metastasis. BRAF V600E mutation has been extensively studied in relation to negative prognostic indicators in PTC, with no consistent relationship emerging. Two recent meta-analyses showed an overall association between BRAF status and aggressive disease features and called for tailoring treatment plans in patients accordingly. In this, the largest US study to date, BRAF status was not significantly associated with most clinicopathologic features suggestive of more aggressive disease.

Utilization of a MAB for BRAFV600E detection in papillary thyroid carcinoma

Identification of BRAF(V600E) in thyroid neoplasia may be useful because it is specific for malignancy, connotes a worse prognosis, and is the target of novel therapies currently under investigation. Sanger sequencing is the 'gold standard' for mutation detection but is subject to sampling error and requires resources beyond many diagnostic pathology laboratories. In this study, we compared immunohistochemistry (IHC) using a BRAF(V600E) mutation-specific MAB to Sanger sequencing on DNA from formalin-fixed paraffin-embedded tissue, in a well-characterized cohort of 101 papillary thyroid carcinoma (PTC) patients. For all cases, an IHC result was available; however, five cases failed Sanger sequencing. Of the 96 cases with molecular data, 68 (71%) were BRAF(V600E) positive by IHC and 59 (61%) were BRAF(V600E) positive by sequencing. Eleven cases were discordant. One case was negative by IHC and initially positive by sequencing. Repeat sequencing of that sample and sequencing of a macrodissected sample were negative for BRAF(V600E). Of ten cases positive by IHC but negative by sequencing on whole sections, repeat sequencing on macrodissected tissue confirmed the IHC result in seven cases (suggesting that these were false negatives of sequencing on whole sections). In three cases, repeat sequencing on recut tissue remained negative (including using massive parallel sequencing), but these cases demonstrated relatively low neoplastic cellularity. We conclude that IHC for BRAF(V600E) is more sensitive and specific than Sanger sequencing in the routine diagnostic setting and may represent the new gold standard for detection of BRAF(V600E) mutation in PTC.

The prognostic significance of the V600E B-Raf mutation in papillary thyroid carcinoma: data from an Australian population

Aims B-Raf V600E mutation in the thyroid is a specific marker of papillary thyroid carcinoma (PTC) and correlates with poorer survival and rates of recurrence.1,2 We assessed the prevalence of this mutation in a series of PTCs from an Australian population, its association with aggressive clinicopathological features, recurrence and mortality. Methods 82 patients with a median follow up of 6 years were studied. V600E mutation status was determined by PCR on DNA extracted from paraffin blocks. Clinicopathological parameters, recurrence and mortality were analysed according to V600E mutation status. Results The V600E mutation was demonstrated in 44 of 82 (54%) tumours. Up to 58% of patients with V600E positive PTCs suffered a recurrence compared to 12% of patients with V600E negative PTCs (p = 0.005). Up to 15% of patients with V600E positive PTCs died during the follow-up period compared to 5% of patients without the mutation (p = 0.19). The V600E mutation was associated with male sex (p = 0.0002) and the classic subtype (p = 0.005). Discussion The prevalence of B-Raf V600E mutation in PTCs in this study is similar to other series. We confirmed that this mutation was associated with a poorer prognosis, specifically higher rates of recurrence. B-Raf V600E mutation in the thyroid is a specific marker of papillary thyroid carcinoma (PTC) and correlates with poorer survival and rates of recurrence.1,2 We assessed the prevalence of this mutation in a series of PTCs from an Australian population, its association with aggressive clinicopathological features, recurrence and mortality. 82 patients with a median follow up of 6 years were studied. V600E mutation status was determined by PCR on DNA extracted from paraffin blocks. Clinicopathological parameters, recurrence and mortality were analysed according to V600E mutation status. The V600E mutation was demonstrated in 44 of 82 (54%) tumours. Up to 58% of patients with V600E positive PTCs suffered a recurrence compared to 12% of patients with V600E negative PTCs (p = 0.005). Up to 15% of patients with V600E positive PTCs died during the follow-up period compared to 5% of patients without the mutation (p = 0.19). The V600E mutation was associated with male sex (p = 0.0002) and the classic subtype (p = 0.005). The prevalence of B-Raf V600E mutation in PTCs in this study is similar to other series. We confirmed that this mutation was associated with a poorer prognosis, specifically higher rates of recurrence.

The prognostic significance of the V600E B-Raf mutation in papillary thyroid carcinoma: data from an Australian population

Aims B-Raf V600E mutation in the thyroid is a specific marker of papillary thyroid carcinoma (PTC) and correlates with poorer survival and rates of recurrence. 1 , 2 We assessed the prevalence of this mutation in a series of PTCs from an Australian population, its association with aggressive clinicopathological features, recurrence and mortality. Methods 82 patients with a median follow up of 6 years were studied. V600E mutation status was determined by PCR on DNA extracted from paraffin blocks. Clinicopathological parameters, recurrence and mortality were analysed according to V600E mutation status. Results The V600E mutation was demonstrated in 44 of 82 (54%) tumours. Up to 58% of patients with V600E positive PTCs suffered a recurrence compared to 12% of patients with V600E negative PTCs ( p =0.005). Up to 15% of patients with V600E positive PTCs died during the follow-up period compared to 5% of patients without the mutation ( p =0.19). The V600E mutation was associated with male sex ( p =0.0002) and the classic subtype ( p =0.005). Discussion The prevalence of B-Raf V600E mutation in PTCs in this study is similar to other series. We confirmed that this mutation was associated with a poorer prognosis, specifically higher rates of recurrence.