Normal Thyroid Tissue Research Articles

Integrated metabolic and genetic analysis reveals distinct features of human differentiated thyroid cancer.

Differentiated thyroid cancer (DTC) affects thousands of lives worldwide each year. Typically, DTC is a treatable disease with a good prognosis. Yet, some patients are subjected to partial or total thyroidectomy and radioiodine therapy to prevent local disease recurrence and metastasis. Unfortunately, thyroidectomy and/or radioiodine therapy often worsen(s) quality of life and might be unnecessary in indolent DTC cases. On the other hand, the lack of biomarkers indicating a potential metastatic thyroid cancer imposes an additional challenge to managing and treating patients with this disease. The presented clinical setting highlights the unmet need for a precise molecular diagnosis of DTC and potential metastatic disease, which should dictate appropriate therapy. In this article, we present a differential multi-omics model approach, including metabolomics, genomics, and bioinformatic models, to distinguish normal glands from thyroid tumours. Additionally, we are proposing biomarkers that could indicate potential metastatic diseases in papillary thyroid cancer (PTC), a sub-class of DTC. Normal and tumour thyroid tissue from DTC patients had a distinct yet well-defined metabolic profile with high levels of anabolic metabolites and/or other metabolites associated with the energy maintenance of tumour cells. The consistency of the DTC metabolic profile allowed us to build a bioinformatic classification model capable of clearly distinguishing normal from tumor thyroid tissues, which might help diagnose thyroid cancer. Moreover, based on PTC patient samples, our data suggest that elevated nuclear and mitochondrial DNA mutational burden, intra-tumour heterogeneity, shortened telomere length, and altered metabolic profile reflect the potential for metastatic disease. Altogether, this work indicates that a differential and integrated multi-omics approach might improve DTC management, perhaps preventing unnecessary thyroid gland removal and/or radioiodine therapy. Well-designed, prospective translational clinical trials will ultimately show the value of this integrated multi-omics approach and early diagnosis of DTC and potential metastatic PTC.

Thyroglobulin measurement in the needle washout for diagnosis of lymph node metastases of differentiated thyroid cancer

Introduction. Measurement of thyroglobulin (Tg) in a washout after fine-needle aspiration (FNA) is recommended for the diagnosis of metastases of differentiated thyroid cancer (DTC), but the method is not standardized and there is no recommended threshold value of Tg washout, which makes it difficult to interpret the results.Aim. To analyze Tg in the washout after FNA of lesions of different origin on the neck and to determine its optimal cutoff for the diagnosis of DTC metastases.Materials and methods. Ultrasound-guided FNA was performed in 1258 neck masses from 591 patients, 566 of them with confirmed DTC. In 1023 lesions, FNA was performed after thyroidectomy, 22 – after lobectomy, 213 – with preserved thyroid gland. The needle after FNA was washed in 1 ml of saline, in which the concentration of Tg was then examined. The level of Tg in the washout was compared with the histological (n = 522) or cytological diagnosis if no surgery was performed. The cut-off for Tg washout was determined by ROC analysis.Results. DTC lymph node metastases detected in 577 specimens. Nine specimens were obtained from metastases of thyroid cancer (TC) that does not express Tg (anaplastic TC, poorly differentiated TC, columnar-cell variant DTC), 22 – from neck metastases of other malignancy (lung cancer, mucinous soft tissues tumor, ovarian cancer, esophageal cancer, melanoma, neuroendocrine tumor), 6 – from other tumors of the neck (lymphoma, parathyroid adenoma, neurinoma). In 26 cases, the specimen was regarded as normal thyroid tissue left after thyroidectomy, 37 – postoperative seroma or granuloma, 1 – cyst of the neck, 578 – lymph node hyperplasia, 2 – sarcoidosis. The level of Tg washout from DTC metastasis and thyroid remnant significantly differed from that of non-thyroidal origin (p <0.0001). At the cut-off of 7.8 ng/ml, the sensitivity and specificity of Tg washout in the diagnosis of DTC metastases is 94 and 95 %, and at the cut-off of 20 ng/ml, 90 and 98 %, respectively. False-negative results were obtained from DTC with squamous metaplasia or sparse tumor cells in a specimen. False-positive results were obtained more often from lesions of level VI and IV compared with other localizations (8 % versus 4 %; p = 0.04). There were no differences in false positive rate in patients before and after thyroidectomy (p = 0.17), but in patients after thyroidectomy with a serum Tg >200 ng/ml, the false positive rate of Tg washout was significantly higher than that with a lower level of serum Tg (28 % versus 3 %; p = 0.0004). When comparing diagnostic performance of cytology and Tg washout, the advantage of the latter is in the diagnosis of cystic metastases, and the former is in the diagnosis of micrometastases and tumors that do not express Tg. Thyroglobulin in the washout increased the sensitivity of the cytology by 8 %. The combined use of these methods detected DTC metastases in 100 % of patients.Conclusion. Measurement of Tg in the washout is a useful addition to the cytology, increasing the diagnostic performance of the latter, mainly due to better detection of cystic metastases of DTC. The optimal suggested cut-off for Tg washout is 20 ng/mL, at which there are fewer false positives.

Clinical and Pathological Features and Gene Expression Profiles of Clinically Aggressive Papillary Thyroid Carcinomas

Papillary thyroid carcinoma (PTC) is considered an indolent neoplasm but it may demonstrate aggressive behavior. We aimed to identify clinical and pathological characteristics and molecular signatures associated with aggressive forms of PTCs. We selected 43 aggressive PTC cases based on the presence of metastases at the time of diagnosis, the development of distant metastasis during follow-up, and/or biochemical recurrence, and 43 PTC patients that were disease-free upon follow-up, matching them according to age, sex, pT, and pN parameters. Twenty-four pairs (a total of 48 cases) and 6 normal thyroid tissues were studied using targeted mRNA screening of cancer-associated genes employing NanoString nCounter® technology. In general, aggressive PTCs showed distinctive clinical and morphological features. Among adverse prognostic parameters, the presence of necrosis and an increased mitotic index were associated with shorter disease-free and overall survivals. Other parameters associated with shorter disease-free or overall survivals include a lack of tumor capsule, the presence of vascular invasion, tumor-infiltrating lymphocytes, fibrosclerotic changes, age > 55 years, and a high pTN stage. Various pathways were differentially regulated in non-aggressive as compared to aggressive PTC, including the DNA damage repair, the MAPK, and the RAS pathways. In particular, the hedgehog pathway was differentially de-regulated in aggressive PTC as compared to non-aggressive PTC cases, being WNT10A and GLI3 genes significantly up- and down-regulated in aggressive PTC and GSK3B up-regulated in non-aggressive PTC cases. In conclusion, our study revealed specific molecular signatures and morphological features in aggressive PTC that may be useful to predict more aggressive behavior in a subset of PTC patients. These findings may be useful when developing novel, tailored treatment options for these patients.

Voltage-Gated Sodium Channel NaV1.7 Inhibitors with Potent Anticancer Activities in Medullary Thyroid Cancer Cells.

Our results from quantitative RT-PCR, Western blotting, immunohistochemistry, and the tissue microarray of medullary thyroid cancer (MTC) cell lines and patient specimens confirm that VGSC subtype NaV1.7 is uniquely expressed in aggressive MTC and not expressed in normal thyroid cells and tissues. We establish the druggability of NaV1.7 in MTC by identifying a novel inhibitor (SV188) and investigate its mode of binding and ability to inhibit INa current in NaV1.7. The whole-cell patch-clamp studies of the SV188 in the NaV1.7 channels expressed in HEK-293 cells show that SV188 inhibited the INa current in NaV1.7 with an IC50 value of 3.6 µM by a voltage- and use-dependent blockade mechanism, and the maximum inhibitory effect is observed when the channel is open. SV188 inhibited the viability of MTC cell lines, MZ-CRC-1 and TT, with IC50 values of 8.47 μM and 9.32 μM, respectively, and significantly inhibited the invasion of MZ-CRC-1 cells by 35% and 52% at 3 μM and 6 μM, respectively. In contrast, SV188 had no effect on the invasion of TT cells derived from primary tumor, which have lower basal expression of NaV1.7. In addition, SV188 at 3 μM significantly inhibited the migration of MZ-CRC-1 and TT cells by 27% and 57%, respectively.

VueBox® perfusion analysis of dynamic contrast enhanced ultrasound provides added value in the diagnosis of small thyroid nodules.

To explore the potential added value of dynamic contrast enhanced ultrasound (DCE-US) using VueBox® software for the diagnosis of small solid thyroid nodules (≤1.0 cm). This prospective study was approved by the institutional review board and it was performed at two hospitals from January 2020 to October 2020. B mode ultrasound and contrast enhanced ultrasound (CEUS) images were obtained for 79 small solid thyroid nodules (≤1.0 cm) confirmed by ultrasound-guided fine needle aspiration cytology results in 79 consecutive patients (55 women and 24 men, median age: 41 years). The CEUS time-intensity curves (TICs) of thyroid nodules and surrounding parenchyma were created by VueBox® software (Bracco, Italy). The CEUS quantitative parameters were obtained after curve fitting. The diagnostic performance of CEUS and DCE-US was evaluated and compared. The weighted kappa statistic (κ) was performed to assess the interobserver agreement and consistency between the diagnosis of CEUS and DCE-US. Among the 79 thyroid nodules, 56 (70.9 %) were malignant and 23 (29.1 %) were benign lesions. Hypoenhancement during the arterial phase of CEUS was associated with malignancy (P < 0.001), with an AUC of 0.705 (sensitivity 71.4 %, specificity 69.6 %). Among all CEUS quantitative parameters, the peak enhancement (PE), wash-in rate (WiR), and wash-out rate (WoR) of DCE-US in malignancies were significantly lower than those in benign nodules (P = 0.049, P = 0.046, and P = 0.020, respectively). The area under the receiver operating characteristic curves (AUCs) of PE, WiR, and WoR were 0.642 (sensitivity 65.2 %, specificity 67.9 %), 0.643 (sensitivity 43.5 %, specificity 91.1 %), and 0.667 (sensitivity 69.6 %, specificity 69.6 %) in differentiation between benign and malignant small solid thyroid nodules (≤1.0 cm), respectively. Comparing the quantitative parameters of DCE-US between small solid thyroid nodules and surrounding normal thyroid parenchyma, the PE, WiAUC, WiR, wash-in perfusion index (WiPI), WoAUC, WiWoAUC, and WoR of the nodules were significantly lower than those of normal thyroid tissue (P = 0.008, P < 0.001, P = 0.037, P = 0.009, P = 0.003, P = 0.002, P = 0.049, respectively). A total of 16 (20.3 %) nodules showed isoenhancement during the arterial phase of CEUS, while the median PE ratio of surrounding tissue and thyroid nodules was 1.70 (IQR: 1.33-1.89). VueBox® is a helpful tool for the evaluation of dynamic microvascularization of thyroid nodules, and DCE-US using VueBox® perfusion analysis could provide added values for differential diagnosis of small solid thyroid nodules (≤1.0 cm).

Three-dimensional structural analysis of papillary thyroid carcinoma nuclei with serial block-face scanning electron microscopy (SBF-SEM).

Nuclear morphology of carcinoma cells is critical for the pathological diagnosis of papillary thyroid carcinoma (PTC). However, three-dimensional architecture of PTC nuclei is still elusive. In this study, we analyzed the three-dimensional ultrastructure of PTC nuclei using serial block-face scanning electron microscopy which takes advantage of the high-throughput acquisition of serial electron microscopic images and three-dimensional reconstruction of subcellular structures. En bloc-stained and resin-embedded specimens were prepared from surgically removed PTCs and normal thyroid tissues. We acquired two-dimensional images from serial block-face scanning electron microscopy and reconstructed three-dimensional nuclear structures. Quantitative comparisons showed that the nuclei of carcinoma cells were larger and more complex than those of normal follicular cells. The three-dimensional reconstruction of carcinoma nuclei divided intranuclear cytoplasmic inclusions into "open intranuclear cytoplasmic inclusions" connecting to cytoplasm outside the nucleus and "closed intranuclear cytoplasmic inclusions" without that connection. Cytoplasm with abundant organelles was observed in open inclusions, but closed inclusions contained fewer organelles with or without degeneration. Granules with a dense core were only observed in closed inclusions. Our observations suggested that open inclusions originate from nuclear invaginations, and disconnection from cytoplasm leads to closed inclusions.

Abstract #1398160: A Rare Case of Water Clear Cell Parathyroid Adenoma Escaping Sestamibi Scan and Neck Ultrasound – A Diagnostic Conundrum of a Rare Pathological Intity with a Rare Presentation

Parathyroid water-clear cell hyperplasia (WCCH) and water-clear cell adenoma (WCCA) are extremely rare causes of primary hyperparathyroidism (PHPT). WCCH contributes to < 1% of all PHPT however WCCA is exceptionally rare and has been reported only in less than 30 case reports. We are presenting a very rare indolent pathological entity with a very rare presentation escaping common diagnostic measures of Sestamibi scan and neck ultrasound. A 63-year-old female presented for evaluation of severe osteoporosis with a T-score of -3.6. Laboratory workup revealed elevated parathyroid hormone (PTH) of 122 pg/ml (reference range 12-88 pg/ml), Normal calcium 9.8 mg/dl, Normal vitamin D 50 ng/ml, and normal magnesium 2.2 mg/dl. 24-hour urine calcium was 421 mg/24H. Diagnosis was consistent with normocalcemic PHPT and meeting surgical criteria with osteoporosis and high urine calcium. Sestamibi scan did not identify any focal activity. CT H&N revealed an 8 mm mass inferior and posterior to right thyroid lobe with partial washout and slightly different enhancement compared to thyroid gland. Neck ultrasound (US) reported that previously noted tissue activity on the CT H&N is not visualized and most likely represents normal thyroid tissue. Patient was referred to otolaryngology surgery and underwent four gland explorations, including right inferior and left superior parathyroidectomy. Pathology of superior parathyroid gland revealed parathyroid water clear cell adenoma. 3-month Postoperative PTH 29.4 pg/ml and calcium level of 9.3 mg/dl. Solitary parathyroid adenoma (SPA) attributes to 80-85% of PHPT. Majority of cases is due to Chief cell adenomas, less frequent pathologies include oxyphil cell adenoma, lipoadenoma, WCCA or mixed type. Scientific literature regarding WCCA presentation and diagnosis is blurred due to its rarity. Clinical presentation of WCCA resembles other etiologies of PHPT, though some data suggest that is frequently indolent and manifests with mild PHPT. Literature suggests that WCCA has low biochemical activity and most cases had normal to mildly elevated calcium-our patient support this hypothesis-. Hypercalcemia does not manifest until significant growth of the adenoma, though few reported cases manifested with hypercalcemia. Unlike other SPA where Sestamibi scan sensitivity is 91%, literature suggests possibility of false-negative results with WCCA which is hypothesized to be attributed to its different histology of glycogen-rich vacuolized cytoplasm that leads to fast Sestamibi wash-out, for the same reason data suggest that US is more decisive given high water content of cells -Our patient didn’t follow this rule and escaped Sistamibi and US though seen on CT- which still makes the diagnosis of WCCA a conundrum. Some cases underwent surgical excision based on clinical diagnosis which led to revision of PHPT and/or hypercalcemia. There is no available data suggesting different long-term follow-up regimens for WCCA compared to other SPA. Further literature with longer follow-up is needed for this extremely rare entity for a comprehensive understanding of its natural course.

Upregulation of dendrocyte-expressed seven transmembrane protein is associated with unfavorable outcomes in differentiated thyroid cancer.

Dendritic cell infiltrates are increased in thyroid cancer but may have a defective ability to provoke effective immune responses. In this study, we aimed to identify potential thyroid cancer biomarkers linked to dendritic cell development and evaluate their prognostic relevance. Through a bioinformatics search, we identified the dendrocyte-expressed seven transmembrane protein (DCSTAMP) as a prognostic gene involved in dendritic cell differentiation for thyroid cancer. Immunohistochemical analyses of DCSTAMP expression were performed and correlated with clinical outcomes. DCSTAMP was overexpressed in a variety of types of thyroid cancers, while normal thyroid tissue or benign thyroid lesions exhibited low or undetectable DCSTAMP immunoreactivity. The results of automated quantification were consistent with subjective semiquantitative scoring. Among 144 patients with differentiated thyroid cancer, high DCSTAMP expression was associated with papillary tumor type (p < 0.001), extrathyroidal extension (p = 0.007), lymph node metastasis (p < 0.001), and BRAF V600E mutation (p = 0.029). Patients with tumors showing high DCSTAMP expression had shorter overall (p = 0.027) and recurrence-free (p = 0.042) survival. This study provides the first evidence of DCSTAMP overexpression in thyroid cancer. Apart from the prognostic implications, studies are needed to explore its potential immunomodulatory role in thyroid cancer.

Abstract 3915: RNAseq analysis reveals differentiation gene drivers as potential therapeutic targets in anaplastic thyroid cancer

Abstract Papillary thyroid cancer (PTC) is a well-differentiated and highly treatable cancer with a 5-year survival rate of almost 99%. In contrast, anaplastic thyroid cancer (ATC) lacks differentiation and loses its thyroid-like functions, including lack of response to radioiodine therapy. Due to ATC’s aggressive metastatic phenotype, it is always diagnosed as stage IV with a 5 year survival rate of less than 3%. ATC lacks targetable genetic lesions making it a difficult cancer to treat, therefore RNAseq analysis was performed in an effort to investigate the differentially expressed genes between ATC to PTC. Exploration of differential gene expression between ATC and PTC was undertaken to further elucidate the differences in expression that drive the aggressive nature of ATC phenotype. RNA was isolated from human thyroid cancer cell lines T238 (ATC) and K1 (PTC) and RNA sequencing was conducted by Azenta Life Sciences. Our analysis resulted in 4,931 significant differentially expressed genes between ATC and PTC. Looking at the gene ontology analysis, pathways involving cell differentiation, proliferation, and migration revealed 90, 70, and 47 genes differentially expressed in ATC vs PTC respectively. These pathways putatively give rise to ATC’s aggressive phenotype when compared to PTC, therefore, we narrowed down the focus from 4,931 genes to 207. Through RNAseq analysis, Homeobox D4 (HOXD4) was observed to significantly increase in ATC (7.69 log2 fold change) compared to PTC. HOXD4 is a transcription factor that plays a crucial role in cell differentiation and has been associated with a worse prognosis in cancers such as ovarian serous carcinoma. As loss of differentiation is essential to ATC’s refractory nature and therapeutic resistance, targeting pathways involved in differentiation can be crucial in making ATC amenable to therapy. Other differentiation regulatory genes identified include BLK and LAMA5. BLK (3.95 log2 fold change) is a Src family tyrosine kinase proto-oncogene that is not only involved in cell proliferation but also differentiation. LAMA5 (4.76 log2 fold change) is an extracellular glycoprotein that has been linked to differentiation, migration and metastasis. Validation of the RNAseq results was completed via western blot analysis of multiple ATC and PTC cells lines. We found that HOXD4 expression was higher in ATC cell lines T238, 8505C, and SW1736 than PTC cell lines K1, TPC1, and BCPAP. Further validation was carried out by immunohistochemistry of human thyroid tissue. Preliminary IHC on ATC, PTC, and normal thyroid patient samples showed ATC has significantly increased HOXD4 expression (27.5%) compared to PTC (8.7%) and normal thyroid tissue (2.3%). Ongoing studies in the laboratory to identify novel genetic drivers of cell differentiation and proliferation so as to discover a therapeutic target to fulfill an unmet clinical need is underway. Citation Format: Kaci Kopec, Tara Jarboe, Michael Guo, Nicole R. DeSouza, Sarnath Singh, Augustine Moscatello, Humayun K. Islam, Jan Geliebter, Raj K. Tiwari. RNAseq analysis reveals differentiation gene drivers as potential therapeutic targets in anaplastic thyroid cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3915.

Abstract 1406: Long non-coding RNA DUXAP10 is a putative determinant of anaplastic thyroid cancer

Abstract Anaplastic Thyroid Cancer (ATC) is one of the most aggressive forms of cancer, with only a 4% survival rate with metastatic disease. ATC is highly refractory and resistant to both traditional therapeutic modalities and recently discovered small molecule inhibitors. The elucidation of molecular mechanisms that contribute to ATC’s metastatic propensity, and the identification of molecular biomarkers for early diagnosis, prognosis, and therapeutic intervention are critical unmet clinical needs-highlighting the importance of novel avenues of intervention. Recently, long non-coding RNAs (lncRNA) have been described as regulators of gene expression and cancer cell phenotypes, exhibiting the importance of identifying non-coding profiles that potentially impact ATC behavior. Publicly available datasets provided by Gene Expression Omnibus (GSE33630, GSE85457) enabled investigation of gene expression profiles of ATC vs. normal thyroid patient tissue. Bioinformatic analyses with the GEO2R program identified the lncRNA, Double Homeobox A Pseudogene 10 (DUXAP10), to be highly upregulated in ATC patient-derived tissue samples when compared to normal thyroid tissue by about 5logFC. Analysis using Gene Expression Profiling Interactive Analysis shows that high DUXAP10 expression is correlated with decreased survival in thyroid cancer patients. To recapitulate this patient data in an in vitro model, we found that the expression of DUXAP10 is also upregulated in the ATC cell line, T238, in comparison to Nthy-ori-3-1, an immortalized “normal” thyroid cell line. To understand the consequences of DUXAP10 overexpression in ATC, we used CRISPRi technology to determine the phenotypic and genotypic effects of DUXAP10 downregulation. CRISPRi knockdown of DUXAP10 significantly reduces the migratory capabilities of T238 by 50%, invasiveness by 37%, clonogenicity by 70%, cell size by 34%, and proliferation by 28%, 45%, and 36% at 24, 48, and 72 hours, respectively. This provides evidence that elevated DUXAP10 expression may increase the probability and negative consequences of metastases. Thus, targeting DUXAP10 expression may have beneficial clinical outcomes. Identification of the molecular innervations of DUXAP10 that orchestrate these pro-metastatic phenotypic changes are currently in progress. Immediate next steps are aimed toward determining if DUXAP10 can be utilized as a potential biomarker for ATC diagnosis and prognosis, as well as elucidating the full range of interactions that DUXAP10 has in shaping the ATC phenotype. Citation Format: Nicole R. DeSouza, Michelle Carnazza, Danielle Quaranto, Tara Jarboe, Kaci Kopec, Sarnath Singh, Augustine Moscatello, Jan Geliebter, Raj K. Tiwari. Long non-coding RNA DUXAP10 is a putative determinant of anaplastic thyroid cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1406.

Potentially Important Markers in Thyroid Neoplasia: Claudin-1 and MMP-7

ABSTRACT Background: Thyroid carcinomas are the most common malignant endocrine tumors, and various immunohistochemical markers are tested in routine practice to reduce diagnostic differences, as well as to elucidate carcinogenesis and detect malignancy. Disruption of basement membranes and the extracellular matrix is an important step in tumor carcinogenesis and progression. The claudin and matrix metalloproteinase families are also thought to be effective in this process. Aim: In this retrospective study, the comparative expression of claudin-1 and MMP-7 immunomarkers in normal tissues and thyroid neoplasia were investigated. Materials and Methods: Immunohistochemical staining was performed for claudin-1 and matrix metalloproteinase 7 (MMP-7) in 112 sections, including 24 follicular adenomas, 22 follicular carcinomas, 24 medullary carcinomas, 24 papillary carcinomas, and 18 single dominant nodules from thyroid lesions. Results: A significant staining difference for claudin-1 was observed in follicular carcinoma and medullary carcinoma, papillary carcinoma, and single dominant nodules compared to normal thyroid tissue. A statistically significant staining difference was observed for MMP-7 in follicular adenoma, medullary carcinoma, and papillary carcinoma compared to normal thyroid tissue. Conclusions: These results indicate that claudin-1 and MMP-7 are important in the diagnosis, differential diagnosis, and carcinogenesis of follicular adenoma, follicular carcinoma, medullary carcinoma, papillary carcinoma, and single dominant nodules.

Thyroid metastasis from cervical carcinoma

There have been very few reports of thyroid metastasis from cervical carcinoma. This article reports a case of metastasis from cervical carcinoma. A 69-year-old woman with a history of cervical cancer presented to our department with right cervical swelling. She had suffered from paralysis of the left recurrent laryngeal nerve after undergoing left hemithyroidectomy 16 years ago. Computed Tomography (CT) and magnetic resonance imaging scans revealed a right thyroid tumor with bilateral enlargement of the cervical lymph nodes. Fluorodeoxyglucose positron emission tomography/CT imaging revealed high uptake in the right thyroid tumor, cervical lymph nodes, and right iliac bone. Fine-needle aspiration cytology revealed atypical epithelial cells with a high nuclear/cell ratio and hyperchromatic nuclei, which resembled cervical cancer cells. Given the risk of airway obstruction and dysphagia, the patient underwent tracheostomy, right hemithyroidectomy, and neck dissection for preserving the quality of life. Pathological examination revealed normal thyroid tissue with p16-positive atypical epithelial cells, which suggested thyroid metastasis from the cervical cancer. The patient has remained alive without any symptoms for 5 postoperative months.

Assessment of Thyroid Stiffness and Viscosity in Autoimmune Thyroiditis Using Novel Ultrasound-Based Techniques

The estimation of viscosity by measuring the shear-wave dispersion (SWD) using ultrasound 2D shear-wave elastography (SWE) is becoming more and more popular. Recent research suggests that SWD can be used in addition to 2D-SWE (shear-wave speed) to diagnose diffuse liver disease. Viscosity was studied for the assessment of normal thyroid tissue. This study aims to evaluate the use of viscosity measurements in patients with chronic autoimmune thyroiditis using the SuperSonic MACH®30 ultrasound machine (Hologic SuperSonic Imagine, Aix-en-Provence, France) which provides the Vi PLUS mode for viscosity and the 2D SWE PLUS mode for shear-wave speed measurements. Valid measurements were obtained in 308 cases, 153 with chronic autoimmune thyroiditis (CAT) and 155 with no thyroid pathology (95.95% feasibility of the methods). The differences between the healthy group and the CAT group were statistically significant both for Vi PLUS (2.5 ± 0.4 vs. 2.8 ± 0.5, p < 0.0001) and for 2D-SWE PLUS (13.5 ± 3.3 vs. 23.1 ± 8.3, p < 0.0001). The diagnostic performance was poor for Vi PLUS alone (AUC = 0.69; cut-off > 2.5 Pa·s, se = 68.6%; sp = 64.52%) and good for 2D-SWE PLUS alone (AUC = 0.861; cut-off > 18.4 kPa, se = 69.9%; sp = 92.2%). Vi PLUS correlated with 2D-SWE PLUS, with the presence of CAT, the thyroid volume, levothyroxine replacement therapy and age. Statistically significant differences were found between the CAT subgroup receiving thyroid replacement therapy and the subgroup without therapy: 24.74 ± 8.33 vs. 21.93 ± 8.12 kPa for 2D-SWE (p = 0.0380) and 3 ± 0.5 vs. 2.7 ± 0.4 Pa·s for Vi PLUS (p = 0.0193). Elastography-based methods improve the classic ultrasound evaluation: 2D-SWE PLUS performed somewhat better in distinguishing CAT from normal thyroid tissue, while Vi PLUS made a slightly better assessment regarding the functional status.

Expression and Clinical Significance of Golgi Phosphoprotein 3 (GOLPH3) in Papillary Thyroid Carcinoma.

This study aimed to explore the correlation of Golgi phosphoprotein 3 (GOLPH3) levels in papillary thyroid carcinoma (PTC) and papillary thyroid microcarcinoma (PTMC) with clinicopathologic features. GOLPH3 expression was determined by western blotting in solid tumors and the adjacent normal thyroid tissues. Mammalian target of rapamycin (mTOR) and Ki-67 were examined by immunohistochemical staining. Significantly higher levels of GOLPH3 protein were observed in PTC and PTMC compared with the adjacent normal thyroid tissues (P<0.001). GOLPH3 level was positively associated with lymph node metastasis and clinical stage in PTC (P<0.05) and utterly related to the clinical stage in PTMC (P=0.012). No correlation was observed between GOLPH3 level and other clinicopathologic parameters such as sex, local invasion, tumor number, and tumor size. The expression level of GOLPH3 protein in mTOR-positive PTC was significantly higher than in mTOR-negative PTC (P=0.002 in PTC, P=0.022 in PTMC) and positively correlated with Ki-67 proliferation index in PTC via Pearson correlation analysis (r=0.353, P=0.007 in PTC; r=0.583, P<0.001 in PTMC). In conclusion, the relative expression level of GOLPH3 protein was significantly higher in PTC and PTMC than in normal thyroid tissues and increased with cancer severity. It may provide adjunctive information for diagnosing and predicting prognosis in patients with PTC or PTMC.

Mitochondrial Ribosomal Protein L14 Promotes Cell Growth and Invasion by Modulating Reactive Oxygen Species in Thyroid Cancer.

The mitochondrial ribosomal protein L14 (MRPL14) is encoded by a nuclear gene and participates in mitochondrial protein translation. In this study, we aimed to investigate the role of MRPL14 in thyroid cancer. We investigated the association between MRPL14 expression and clinicopathological features using The Cancer Genome Atlas (TCGA) and Chungnam National University Hospital (CNUH) databases. Functional studies of MRPL14, including proliferation, migration, invasion, mitochondrial oxidative phosphorylation and reactive oxygen species (ROS) production, were performed in papillary thyroid cancer (PTC) cell lines (B-CPAP and KTC-1). Based on the TCGA dataset, PTC tissues lost mitochondrial integrity and showed dysregulated expression of overall mitoribosomal proteins (MRPs) compared with normal thyroid tissues. Of 78 MRPs, MRPL14 was highly expressed in thyroid cancer tissues. MRPL14 overexpression was significantly associated with advanced tumor stage, extrathyroidal extension, and lymph node metastasis. MRPL14 increased cell proliferation of thyroid cancer and promoted cell migration via epithelial-mesenchymal transition-related proteins. Moreover, MRPL14 knockdown reduced the expression of oxidative phosphorylation complex IV (MTCO1) and increased the accumulation of ROS. Cotreatment with a ROS scavenger restored cell proliferation and migration, which had been reduced by MRPL14 knockdown, implying that ROS functions as a key regulator of the oncogenic effects of MRPL14 in thyroid cancer cells. Our findings indicate that MRPL14 may promote cell growth, migration, and invasion by modulating ROS in thyroid cancer cells.

Quantitative Iodine-123 single-photon emission computed tomography/computed tomography for Iodine-131 therapy of an autonomously functioning thyroid nodule

PurposeAutonomously functioning thyroid nodules (AFTNs) are treated with iodine-131 (I-131) therapy, which increases the risk of permanent hypothyroidism; however, the risk can be reduced by separately estimating the accumulated activity for the AFTN and extranodular thyroid tissue (ETT).MethodsA quantitative I-123 single-photon emission computed tomography (SPECT)/CT (5 mCi) was performed in one patient with unilateral AFTN and T3 thyrotoxicosis. The I-123 concentrations measured at 24 h were 12.26 µCi/mL and 0.11 µCi/mL in the AFTN and contralateral ETT, respectively. Thus, the I-131 concentrations and radioactive iodine uptake expected at 24 h by 5 mCi of I-131 were 38.59 µCi/mL and 0.31 for the AFTN and 0.34 µCi/mL and 0.007 for the contralateral ETT. The weight was calculated as CT-measured volume multiplied by 1.03.ResultsIn the AFTN patient with thyrotoxicosis, we administered 30 mCi of I-131, which would maximize the 24-h I-131 concentration in the AFTN (226.86 µCi/g) and maintain a tolerable concentration in the ETT (1.97 µCi/g). The percentage of I-131 uptake at 48 h post I-131 administration was 62.6%. The patient achieved a euthyroid state at 14 weeks and maintained the state until 2 years post I-131 administration with an AFTN volume reduction of 61.38%.ConclusionThe pre-therapeutic planning of quantitative I-123 SPECT/CT may enable a therapeutic window for I-131 therapy, which directs optimal I-131 activity to effectively treat AFTN while preserving the normal thyroid tissue.

Single Cell Analysis of Human Thyroid Reveals the Transcriptional Signatures of Aging.

The thyroid gland plays a critical role in the maintenance of whole-body metabolism. However, aging frequently impairs homeostatic maintenance by thyroid hormones due to increased prevalence of subclinical hypothyroidism associated with mitochondrial dysfunction, inflammation, and fibrosis. To understand the specific aging-related changes of endocrine function in thyroid epithelial cells, we performed single-cell RNA sequencing (RNA-seq) of 54 726 cells derived from pathologically normal thyroid tissues from 7 patients who underwent thyroidectomy. Thyroid endocrine epithelial cells were clustered into 5 distinct subpopulations, and a subset of cells was found to be particularly vulnerable with aging, showing functional deterioration associated with the expression of metallothionein (MT) and major histocompatibility complex class II genes. We further validated that increased expression of MT family genes are highly correlated with thyroid gland aging in bulk RNAseq datasets. This study provides evidence that aging induces specific transcriptomic changes across multiple cell populations in the human thyroid gland.

Adding pieces to the puzzle of differentiated-to-anaplastic thyroid cancer evolution: the oncogene E2F7

Anaplastic Thyroid Cancer (ATC) is the most aggressive and de-differentiated subtype of thyroid cancer. Many studies hypothesized that ATC derives from Differentiated Thyroid Carcinoma (DTC) through a de-differentiation process triggered by specific molecular events still largely unknown. E2F7 is an atypical member of the E2F family. Known as cell cycle inhibitor and keeper of genomic stability, in specific contexts its function is oncogenic, guiding cancer progression. We performed a meta-analysis on 279 gene expression profiles, from 8 Gene Expression Omnibus patient samples datasets, to explore the causal relationship between DTC and ATC. We defined 3 specific gene signatures describing the evolution from normal thyroid tissue to DTC and ATC and validated them in a cohort of human surgically resected ATCs collected in our Institution. We identified E2F7 as a key player in the DTC-ATC transition and showed in vitro that its down-regulation reduced ATC cells’ aggressiveness features. RNA-seq and ChIP-seq profiling allowed the identification of the E2F7 specific gene program, which is mainly related to cell cycle progression and DNA repair ability. Overall, this study identified a signature describing DTC de-differentiation toward ATC subtype and unveiled an E2F7-dependent transcriptional program supporting this process.

A rare case of Riedel's thyroiditis: Immunoglobulin G4-related thyroid disease

Abstract Introduction Riedel's thyroiditis is a rare thyroiditis in which normal thyroid tissue is replaced by dense fibrous connective tissue. Its etiology is not fully known. An autoimmune process is thought to exist. In this disease group, lymphoplasmocytic tissue infiltration, mostly consisting of IgG4 (+) plasma cells and small lymphocytes, is seen accompanying fibrosis. In our case, we presented a very rare case of Reidel's thyroiditis. Clinical Case We report the case of a 46-year-old woman who came to the general surgery department with an enlargement at the base of her neck. Among the patient's complaints were dysphonia, a feeling of pressure and pain in the neck. She had no history of thyroid disease. Laboratory findings; TSH 0.06 uIU/ml (0.27–4.2), fT3 2.69 pg/mL (2.3–4.5), fT4 1.81 ng/dl (0.93–1.70), crp 73.6 mg/l (0.0–5.0), sedimentation 90 mm/h (0–20), leukocytes 11.74 10^3/ul (4.49–12.68), absolute neutrophil count 9.22 10 ^3/ul (1.9–7.9). Trucut biopsy was performed with the preliminary diagnosis of malignancy due to the history of a mass growing in a short time on the neck of the patient. Pathology result; “Intense fibrosis increase, severe lymphoplasmacytic cell infiltration rich in IgG4 positive cells. Thyroid follicle epithelial cells with ck7, panck, pax8, thyroglobulin, ttf1 positive and cea, ck20, alk negative Intense plasma cells with CD38 have polyclonal staining with kappa and lambda, and IgG4/IgG ratio &amp;gt; 50%. In the background, cd3, pax5 and bcl2 and T cell-rich reactive inflammatory cells are accompanied. Obliterative vascular structures were noted in rust and elastic histochemical stains, and storiform fibrosis was observed in places on the ground. When histopathological and immunohistochemical findings are evaluated together, it is primarily compatible with IgG4-related disease/reidel thyroiditis.'' She was then referred to the endocrinology department. The serum IgG4 level was found to be above normal. She was started on steroid treatment with 64 mg daily methylprednisolone. A weekly control plan was made. In these controls, the steroid dose was gradually reduced. In the follow-up one week later, it was observed that the patient's CRP and sedimentation returned to normal values. There was also a decrease in enlargement in the neck area. In the third month of the treatment, the patient didn't want to continue the treatment. She did not visit our clinic for the next 3 months. Then, she came for the control after three months without treatment. Biochemistry and thyroid function tests were normal. The swelling in the neck area of the patient was completely gone. Conclusion Reidel's thyroiditis is a very rare disease with publications on the basis of case reports in the literature. Therefore, there are no broad-spectrum clinical studies. Clarity on treatment approaches was also not provided. We also benefited from case reports while managing our patient's treatment. We believe that our case will contribute to the literature for the same purpose.

Ectopic lingual thyroid tissue diagnosed in an elderly patient

Abstract Introduction Lingual thyroid (LT) is a rare ectopy of the gland characterized by residual thyroid tissue remaining along the thyroglossal duct, due to inability of the gland to descend from the foramen cecum into the lower part of the neck. Clinical Case A 65-year-old female patient presented with a long-standing difficulty in swallowing and sensation of a foreign body in the throat. Oropharyngeal examination revealed a well-circumscribed submucosal mass at the base of the tongue. Neck MRI detected 28×25×30 mm, lesion, isointense on T1WI and T2WI, showing prominent and homogeneous contrast enhancement located at the base of tongue, midline at the level of the vallecula, and narrowing towards the oropharynx anteriorly. Since normal thyroid tissue was not observed in the thyroid bed, the mass was suspected to be an ectopic lingual thyroid tissue. Tc99m pertechnetate thyroid scintigraphy revealed focal involvement at the base of the tongue rather than its anatomical region, confirming the diagnosis of LT. Thyroid function tests showed; serum TSH level of 4.25 mU/L (normal range: 0.27–4.20), free T4 12.1 pmol/L (11–22), free T3 4.1 pmol/ L (3.1–6.8), Anti-TPO &amp;lt;0.9 IU/ml, Anti-Tg: 14.9 IU/ml(0–115).Since the patient had difficulty swallowing, Levothyroxine was considered to be insufficient and time-consuming, surgical intervention was planned as a permanent solution. After surgical treatment, LT4 replacement was initiated, and the pathology confirmed normal thyroid tissue. The patient is being followed up with LT4 replacement therapy. Lingual thyroid (LT) is the most frequent ectopic location of the thyroid gland, its prevalence ranges between 1/100000 and 1/300000 being seven times more common in women. While the ectopy is often diagnosed at an early age, it may present at any age from birth to senility. Patients with ectopic LT are usually asymptomatic, hypothyroidism has been reported in 33% to 62% of patients. In proportion to the size of the lingual thyroid tissue, it can also cause complaints such as dysphonia, dysphagia, respiratory distress, foreign body sensation in the throat. Diagnosed with ectopic LT tissue at an advanced age, our patient had complaints of long-standing difficulty in swallowing solid foods, but hypothyroidism was not detected. After ENT examination, the diagnosis was confirmed by technetium (Tc99m) pertechnetate thyroid scintigraphy. Treatment of ectopic LT is either surgical or medical, depending on the size of the mass and patient's signs and symptoms. Ectopic lingual thyroid gland should always be considered, for the midline lesions, especially at the base of the tongue. Detailed assessment should be conducted to conclusively rule out obstruction and malignancy where the surgery is indicated. Levothyroxine could be a good solution for smaller, well defined benign lesions, which may stay stable or reduce in size under treatment.