Int J Thyroidol.  2017 May;10(1):24-35. 10.11106/ijt.2017.10.1.24.

The Cancer Genome Atlas Validation of Ancillary Tests for Classifying Papillary Thyroid Carcinoma

• Suh YJ
• Moon HJ
• Choe JY
• Park HJ

• KMID: 2379358
• DOI: http://doi.org/10.11106/ijt.2017.10.1.24

Abstract

BACKGROUND AND OBJECTIVES
Ancillary tests such as BRAF(V600E) mutation or immunohistochemical (IHC) assays have been utilized as complements to morphological criteria in diagnosing subsets of papillary thyroid carcinoma (PTC). Utilizing results from analysis by The Cancer Genome Atlas (TCGA), we evaluated the diagnostic value and feasibility of these ancillary tests in diagnosing follicular variant PTC (FVPTC).
MATERIALS AND METHODS
Clinical data and tissue samples were analyzed from 370 PTC patients, who had undergone thyroidectomy between December 2003 and July 2014. PTC was limited to conventional PTC (CVPTC), tall cell variant PTC (TCPTC), and FVPTC. Using multivariate analyses, FVPTC cases were compared to CVPTC and TCPTC cases. Surgical specimens were pyrosequenced for BRAF(V600E) mutation or stained for IHC markers such as CD56, galectin-3, cytokeratin 19 (CK19), or CD31. For the validation, a comprehensive analysis was performed for BRAF(V600E) mutation and quantitative mRNA expressional difference in TCGA.
RESULTS
Demographic differences were not observed between 159 CVPTC, 103 TCPTC, and 108 FVPTC cases. BRAF(V600E) mutation predominated in CVPTC+TCPTC group, but not in FVPTC group (78.4% vs. 18.7%, p<0.001), as suggested by TCGA (57.4% vs. 12.1%, p<0.0001). IHC markers significantly distinguished FVPTC cases from CVPTC+TCPTC cases. CD56 exhibited more intense staining in FVPTC cases (21.1% vs. 72.0%, p<0.001). Galectin-3 and CK19 yielded limited values. CD31 correlated with lymphovascular invasion (r=0.847, p<0.001). In analysis of TCGA, mRNA differential expression of these genes revealed their corresponding upregulation or downregulation.
CONCLUSION
BRAF(V600E) mutation or TCGA-validated IHC assay could be recommended as ancillary tests for classifying PTC.

Keyword

BRAF; Immunohistochemistry marker; Thyroid neoplasm; Follicular variant papillary thyroid carcinoma; TCGA

MeSH Terms

Complement System Proteins
Down-Regulation
Galectin 3
Genome*
Humans
Keratin-19
Multivariate Analysis
RNA, Messenger
Thyroid Gland*
Thyroid Neoplasms*
Thyroidectomy
Up-Regulation
Complement System Proteins
Galectin 3
Keratin-19
RNA, Messenger

Figure

• 

  Fig. 1 Study design flow diagram. CVPTC: conventional papillary thyroid carcinoma, FVPTC: follicular variant papillary thyroid carcinoma, ICD: international classification of diseases, IHC: immunohistochemical, TCPTC: tall cell variant papillary thyroid carcinoma

• 

  Fig. 2 Immunohistochemical stains of CD56, Galetin-3, CK19, and CD31, grouped according to papillary thyroid carcinoma histology. CVPTC: conventional papillary thyroid carcinoma, FVPTC: follicular variant papillary thyroid carcinoma, TCPTC: tall cell variant papillary thyroid carcinoma. As per Bioethics and Safety Act in Korea, the slides only within the valid storage period could be suggested in this figure.

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  Fig. 3 (A-D) Boxplots of mRNA immunohistochemical markers grouped according to TCGA histology. CVPTC: conventional papillary thyroid carcinoma, FVPTC: follicular variant papillary thyroid carcinoma, RPKM: reads per kilobase per million formula, TCPTC: tall cell variant papillary thyroid carcinoma

• 

  Fig. 4 (A-D) Quantitative mRNA boxplots of immunohistochemical markers different between CVPTC+TCPTC and FVPTC, in The Cancer Genome Atlas (TCGA). CVPTC: conventional papillary thyroid carcinoma, FVPTC: follicular variant papillary thyroid carcinoma, RPKM: reads per kilobase per million formula, TCPTC: tall cell variant papillary thyroid carcinoma

• 

  Fig. 5 mRNA gene expression differences between follicular variant papillary thyroid carcinoma and normal TCGA samples (Threshold: FDR 25%). (A) Gene Set Enrichment Analysis, (B) Heat map of the top 50 features for each phenotype in gene expression data.

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