Biotin Interference in Assays for Thyroid Hormones, Thyrotropin and Thyroglobulin.

Background: Biotin has been reported to interfere with several commonly used laboratory assays resulting in misleading values and possible erroneous diagnosis and treatment. This report describes a prospective study of possible biotin interference in thyroid-related laboratory assays, with a comparison of different commonly used assay platforms. Materials and Methods: Thirteen adult subjects (mean age 45 ± 13 years old) were administered biotin 10 mg/day for eight days. Blood specimens were collected at three time points on day 1 and on day 8 (baseline, two, and five hours after biotin ingestion). Thyrotropin (TSH), free triiodothyronine (fT3), free thyroxine (fT4), total triiodothyronine (TT3), total thyroxine (TT4), thyroxine binding globulin (TBG), and thyroglobulin (Tg) levels were analyzed with four different platforms: Abbott Architect, Roche Cobas 6000, Siemens IMMULITE 2000, and liquid chromatography with tandem mass spectrometry (LC-MS/MS). TSH, fT3, fT4, TT3, and TT4 were measured with Abbott Architect and Roche Cobas 6000. fT3, fT4, TT3, and TT4 were also measured by LC-MS/MS. Tg was measured by Siemens IMMULITE 2000. TBG was assessed with Siemens IMMULITE 2000. Results: Significant changes in TSH, fT4, and TT3 measurements were observed after biotin exposure when the Roche Cobas 6000 platform was used. Biotin intake resulted in a falsely lower Tg level when measurements were performed with Siemens IMMULITE 2000. At the time points examined, maximal biotin interference was observed two hours after biotin exposure both on day 1 and day 8. Conclusions: A daily dose of 10 mg was shown to interfere with specific assays for TSH, fT4, TT3, and Tg. Physicians must be aware of the potential risk of erroneous test results in subjects taking biotin supplements. Altered test results for TSH and Tg can be particularly problematic in patients requiring careful titration of levothyroxine therapy such as those with thyroid cancer.

Do Molecular Profiles of Primary Versus Metastatic Radioiodine Refractory Differentiated Thyroid Cancer Differ?

Management of metastatic radioiodine refractory differentiated thyroid cancer (DTC) can be a therapeutic challenge. Generally, little is known about the paired molecular profile of the primary tumor and the metastases and whether they harbor the same genetic abnormalities. The present study compared the molecular profile of paired tumor specimens (primary tumor/metastatic sites) from patients with radioiodine refractory DTC in order to gain insight into a possible basis for resistance to radioiodine. Twelve patients with radioiodine refractory metastases were studied; median age at diagnosis of 61 years (range, 25-82). Nine patients had papillary TC (PTC), one had follicular TC (FTC), and two had Hürthle cell TC (HTC). Distant metastases were present in the lungs (n = 10), bones (n = 4), and liver (n = 1). The molecular profiling of paired tumors was performed with a panel of 592 genes for Next Generation Sequencing, RNA-sequencing, and immunohistochemistry. Digital microfluidic PCR was used to investigate TERT promoter mutations. The genetic landscape of all paired sites comprised BRAF, NRAS, HRAS, TP53, ATM, MUTYH, POLE, and NTRK genes, including BRAF and NTRK fusions. BRAF V600E was the most common point mutation in the paired specimens (5/12). TERT promoter mutation C228T was detected in one case. PD-L1 expression at metastatic sites was highly positive (95%) for one patient with HTC. All specimens were stable for microsatellite instability testing, and the tumor mutation burden was low to intermediate. Therefore, the molecular profile of DTC primary and metastatic lesions can show heterogeneity, which may help explain some altered responses to therapeutic intervention.

Cytochrome C Oxidase Subunit 4 (COX4): A Potential Therapeutic Target for the Treatment of Medullary Thyroid Cancer.

The nuclear-encoded subunit 4 of cytochrome c oxidase (COX4) plays a role in regulation of oxidative phosphorylation and contributes to cancer progression. We sought to determine the role of COX4 in differentiated (DTC) and medullary (MTC) thyroid cancers. We examined the expression of COX4 in human thyroid tumors by immunostaining and used shRNA-mediated knockdown of COX4 to evaluate its functional contributions in thyroid cancer cell lines. In human thyroid tissue, the expression of COX4 was higher in cancers than in either normal thyroid (p = 0.0001) or adenomas (p = 0.001). The level of COX4 expression correlated with tumor size (p = 0.04) and lymph-node metastases (p = 0.024) in patients with MTCs. COX4 silencing had no effects on cell signaling activation and mitochondrial respiration in DTC cell lines (FTC133 and BCPAP). In MTC-derived TT cells, COX4 silencing inhibited p70S6K/pS6 and p-ERK signaling, and was associated with decreased oxygen consumption and ATP production. Treatment with potassium cyanide had minimal effects on FTC133 and BCPAP, but inhibited mitochondrial respiration and induced apoptosis in MTC-derived TT cells. Our data demonstrated that metastatic MTCs are characterized by increased expression of COX4, and MTC-derived TT cells are vulnerable to COX4 silencing. These data suggest that COX4 can be considered as a novel molecular target for the treatment of MTC.

Detection of BRAFV600E in Liquid Biopsy from Patients with Papillary Thyroid Cancer Is Associated with Tumor Aggressiveness and Response to Therapy.

The detection of rare mutational targets in plasma (liquid biopsy) has emerged as a promising tool for the assessment of patients with cancer. We determined the presence of cell-free DNA containing the BRAFV600E mutations (cfBRAFV600E) in plasma samples from 57 patients with papillary thyroid cancer (PTC) with somatic BRAFV600E mutation-positive primary tumors using microfluidic digital PCR, and co-amplification at lower denaturation temperature (COLD) PCR. Mutant cfBRAFV600E alleles were detected in 24/57 (42.1%) of the examined patients. The presence of cfBRAFV600E was significantly associated with tumor size (p = 0.03), multifocal patterns of growth (p = 0.03), the presence of extrathyroidal gross extension (p = 0.02) and the presence of pulmonary micrometastases (p = 0.04). In patients with low-, intermediate- and high-risk PTCs, cfBRAFV600E was detected in 4/19 (21.0%), 8/22 (36.3%) and 12/16 (75.0%) of cases, respectively. Patients with detectable cfBRAFV600E were characterized by a 4.68 times higher likelihood of non-excellent response to therapy, as compared to patients without detectable cfBRAFV600E (OR (odds ratios), 4.68; 95% CI (confidence intervals)) 1.26-17.32; p = 0.02). In summary, the combination of digital polymerase chain reaction (dPCR) with COLD-PCR enables the detection of BRAFV600E in the liquid biopsy from patients with PTCs and could prove useful for the identification of patients with PTC at an increased risk for a structurally or biochemically incomplete or indeterminate response to treatment.

Microfluidic Droplet Digital PCR Is a Powerful Tool for Detection of BRAF and TERT Mutations in Papillary Thyroid Carcinomas.

We examined the utility of microfluidic digital PCR (dPCR) for detection of BRAF and TERT mutations in thyroid tumors. DNA extracted from 100 thyroid tumors (10 follicular adenomas, 10 follicular cancers, 5 medullary cancers, and 75 papillary thyroid cancer (PTC) were used for detection of BRAF and TERT mutations. Digital PCRs were performed using rare mutation SNP genotyping assays on QuantStudio 3D platform. In PTCs, BRAFV600E was detected by dPCR and Sanger sequencing in 42/75 (56%) and in 37/75 (49%), respectively. BRAFV600E was not detected in other tumors. The ratio of mutant/total BRAF alleles varied from 4.7% to 47.5%. These ratios were higher in classical PTCs (27.1%) as compared to follicular variant PTCs (9.4%) p = 0.001. In PTCs with and without metastases, the ratios of mutant/total BRAF alleles were 27.6% and 18.4%, respectively, (p = 0.03). In metastatic lesions percentages of mutant/total BRAF alleles were similar to those detected in primary tumors. TERTC228T and TERTC250T were found in two and one cases, respectively, and these tumors concomitantly harbored BRAFV600E. These tumors exhibited gross extra-thyroidal extension, metastases to lymph nodes, and pulmonary metastases (one case). Our results showed that dPCR allows quantitative assessment of druggable targets in PTCs and could be helpful in a molecular-based stratification of prognosis in patients with thyroid cancer.

Evaluation of Thyroid Hormone Replacement Dosing in Overweight and Obese Patients After a Thyroidectomy.

Background: Initiation of thyroid hormone replacement (THR) after a total thyroidectomy has traditionally relied on the weight of the patient, regardless of the patient's body mass index (BMI). Current literature suggests that THR in obese patients differs from nonobese patients. This can lead to overdosing of levothyroxine (LT4) and delay in achievement of euthyroid state. Methods: We retrospectively identified patients on THR after total thyroidectomy with a benign postoperative diagnosis. Patients who achieved euthyroidism with THR were included in the analysis. Patient demographic and THR dosing information was collected. Regression analysis was performed to identify appropriate THR dosing at varying BMIs. This study aimed to evaluate the appropriate dosing of THR in overweight and obese patients. Results: Our cohort consisted of 114 patients achieving euthyroidism while on THR. Mean age was 55 years (range 28-77 years) with 84% females. Of the 114 patients, the number of patients with a BMI less than 25, 25-29, 30-34, 35-39, and greater than 40 were 26 (23%), 33 (29%), 23 (20%), 19 (17%), and 13 (11%), respectively. Of the entire cohort, a mean of 50 weeks elapsed after surgery to achieve euthyroidism, with no significant difference between the BMI categories (p = 0.58). In obese patients (BMI >30), 35% were overdosed with LT4 on initial dosing. The cohort lost a mean of 3 kg until euthyroidism was achieved, with no significant difference in the weight loss based on BMI category (p = 0.61). Patients with a higher BMI did require a higher dose (mcg) of LT4 to achieve euthyroidism (p < 0.01), but the dose was significantly lower in relation to their weight (mcg/kg) (p < 0.01). The LT4 dose required to achieve euthyroidism based on the previously mentioned BMI categories were 1.76, 1.47, 1.42, 1.27, and 1.28 mcg/kg. Conclusion: The current weight-based dosing of THR inappropriately overdoses overweight and obese patients. A more appropriate formula for THR titration should consider both the weight and BMI of the patient.

124I Positron Emission Tomography/Computed Tomography Versus Conventional Radioiodine Imaging in Differentiated Thyroid Cancer: A Review.

Background: Studies report a wide spectrum of 124I positron emission tomography (PET)/computed tomography (CT) sensitivity and specificity in the detection of differentiated thyroid cancer (DTC) lesions. This study reviews the lesion detection rate of pretherapy 124I PET/CT in different patient populations and further analyzes the factors necessary for a better detection on 124I PET/CT. Methods: A literature search was performed using multiple different databases (MEDLINE, EMBASE, Northern Lights, and handsearching) covering 1996 to April 2018. Two reviewers reviewed and extracted study data for 124I, 123I, and 131I scans in DTC. Results: This review includes 4 retrospective and 10 prospective studies in which 495 DTC patients underwent 124I and 131I imaging; no studies made comparisons with 123I. In the reports that compared 124I PET/CT with diagnostic 131I scans, there were a total of 72 patients in whom 120 lesions were detected on 124I imaging, whereas only 52 were detected on diagnostic 131I scans. In publications that compared 124I with post-therapy 131I scans in 266 patients, 410 lesions were detected with 124I PET, whereas 390 were detected on post-therapy 131I scans. Based on 124I PET/CT in six studies, TNM staging was revised in 15-21% of patients, and disease management was altered in 5-29% of patients. Conclusions:124I PET/CT is able to identify a greater number of foci compared with diagnostic 131I scans. 124I PET may have better detection compared with post-therapy 131I scans in patients who are 131I therapy naive, have less aggressive pathology, or do not have disseminated lung metastases. Additional metastatic lesion detection by 124I PET may have a significant clinical impact in the management of patients before 131I therapy in some patients.

Conventional Radioiodine Therapy for Differentiated Thyroid Cancer.

This article presents an overview of the use of radioactive iodine (131-I) in the treatment of patients with differentiated thyroid cancer. Topics reviewed include definitions; staging; the 2 principal methods for selection of 131-I dosage; the indications for ablation, adjuvant treatment, and treatment; the recommendations for the use of 131-I contained in the guidelines of the American Thyroid Association and the Society of Nuclear Medicine and Molecular Imaging; the dosage recommendations and selection of dosage approach for 131-I by these organizations; the use of recombinant human thyrotropin for radioiodine ablation, adjuvant therapy, or treatment; and the MedStar Washington Hospital Center approach.

Use of 99mTc-sestamibi SPECT/CT when conventional imaging studies are negative for localizing suspected recurrence in differentiated thyroid cancer: a method and a lesson for clinical management.

PURPOSE: The detection of recurrent disease in differentiated thyroid cancer (DTC) patients with elevated or rising serum thyroglobulin (Tg) levels and multiple negative conventional imaging studies can be challenging, especially when 18F-FDG PET/CT scan is also negative. We report a patient and review the literature on the diagnostic use of 99mTc-sestamibi scans to identify the source of elevated or rising Tg in patients with negative conventional imaging including negative 18F-FDG PET/CT scans. PATIENT AND METHODS: A 73-year-old woman was referred for widely-invasive metastatic follicular thyroid cancer with bone metastasis to her left mandible. She had a total thyroidectomy, left mandibular resection, and 131I therapy of 145 mCi (5.4 GBq) and her subsequent unstimulated serum Tg level was 29 ng/ml (TgAb negative). At six months' follow-up, her stimulated Tg was 527 ng/ml (TSH 188 mIU/L, TgAb negative). All imaging studies performed within the prior 12 months were reported as negative for recurrence or metastasis; this included neck ultrasound, diagnostic radioiodine scan, chest CT and, 18F-FDG PET/CT. The patient was injected with 24.6 mCi (910 MBq) of 99mTc-sestamibi intravenously, and whole-body and SPECT/CT images were acquired. RESULTS: The 99mTc-sestamibi whole-body posterior image demonstrated abnormal focal uptake in the right posterior calvarium and corresponded to an occipital lytic bone lesion on the SPECT/CT. The patient underwent surgical resection of the skull metastasis, and pathology confirmed metastatic follicular thyroid cancer. Five months post-surgery, the suppressed Tg was markedly reduced and remained stable at ~3.2 ng/ml. With the knowledge of the DTC recurrence location, the two sets of 18F-FDG images were re-evaluated. The more thorough and targeted interpretation underscored the importance of structured image reporting. The current literature on the utility of 99mTc-sestamibi scans when radioiodine, 18F-FDG PET/CT, and other imaging studies are negative is sparse and inconsistent. CONCLUSIONS: 99mTc-sestamibi may have a role in thyroid cancer localization when physical exam, neck ultrasound, radioiodine scan, chest/abdomen CT, and 18F-FDG PET/CT does not identify the source of elevated Tg levels in DTC.

Eliminating the Age Cutoff in Staging of Differentiated Thyroid Cancer: The Safest Road?

Background: Unlike virtually all other cancer types, thyroid cancer is unique in that patient age is a key component in its staging. Pathologists and clinicians worldwide have accepted an age cutoff of 45 years for staging; in 2018, this advances to age 55 years in the eighth edition of the American Joint Commission on Cancer staging system. Evidence Acquisition: Clinical and basic research studies, reviews, and previous editions of consensus statements regarding thyroid cancer staging were reviewed, with particular focus on the influence of age in thyroid cancer prognosis. Purpose: This perspective briefly reviews the basis for this practice and challenges it as no more appropriate than for other malignancies. Evidence Synthesis: The majority of findings report an association of age with thyroid cancer survival but do not support a specific age cutoff; rather, they suggest that outcome is affected by age as a continuous variable. Conceivably, other factors interact with age on a continuous basis over time, affecting prognosis. When identified, these factors could alter our current concept of the importance of an age cutoff in staging. Conclusions: Among all cancers, age has an important role in only thyroid cancer staging. The consideration of age as a continuous variable and the search for age-associated prognostic variables could elucidate a more accurate staging system.