Supplementary Material for: Hyperfunctioning Papillary Thyroid Carcinoma with a <b><i>BRAF</i></b> Mutation: The First Case Report and a Literature Review

<b><i>Introduction:</i></b> Hyperfunctioning papillary thyroid carcinoma (PTC) is rare and consequently, little information on its molecular etiology is available. Although <i>BRAF</i> V600E (<i>BRAF</i> c.1799T&gt;A, p.V600E) is a prominent oncogene in PTC, its mutation has not yet been reported in hyperfunctioning PTC. <b><i>Case Presentation:</i></b> Ultrasonography detected a 26-mm nodule in the right lobe of the thyroid gland of a 48-year-old man. Thyroid function tests indicated that he was hyperthyroid with a TSH level of 0.01 mIU/L (reference range: 0.05–5.00) and a free thyroxine level of 23.2 pmol/L (reference range: 11.6–21.9). TSHR autoantibodies were &lt;0.8 IU/L (reference value: &lt;2.0 IU/L). The ^99mTc thyroid scintigram revealed a round, right-sided focus of tracer uptake by the nodule with a decreased uptake in the remainder of the gland. The patient underwent total thyroidectomy because fine-needle aspiration cytology revealed a malignancy. The histopathological diagnosis was conventional PTC. Subsequent mutational analysis of <i>BRAF</i> (exon 15), <i>TSHR</i> (exons 1–10), <i>GNAS</i> (exons 7–10), <i>EZH1</i> (exon 16), <i>KRAS</i>, <i>NRAS</i>, <i>HRAS</i> (codons 12, 13, and 61), and <i>TERT</i> promoter (C250T and C228T) identified a heterozygous point mutation in <i>BRAF</i> V600E in a tumor tissue sample. In addition, we identified a <i>TSHR</i> D727E polymorphism (<i>TSHR</i> c.2181C&gt;G, p.D727E) in both the tumor and the surrounding normal thyroid tissue. <b><i>Discussion and Conclusions:</i></b> We report a case of hyperfunctioning PTC with a <i>BRAF</i> V600E mutation for the first time. Our literature search yielded 16 cases of hyperfunctioning thyroid carcinoma in which a mutational analysis was conducted. We identified <i>TSHR</i> mutations in 13 of these cases. One case revealed a combination of <i>TSHR</i> and <i>KRAS</i> mutations; the other case revealed a <i>TSHR</i> mutation with a <i>PAX8/PPARG</i> rearrangement. These findings suggest that the concomitant activation of oncogenes (in addition to constitutive activation of the TSHR-cyclic AMP cascade) are associated with the malignant phenotype in hyperfunctioning thyroid nodules.