Marker For Thyroid Cancer Research Articles

Significance of molecular detection in diagnosis of benign and malignant thyroid nodules in China.

e15028 Background: Ultrasound and ultrasound-guided fine needle aspiration (US-FNA) are a preferred method for judging benign and malignant thyroid nodules. For thyroid nodules that can’t be determined by FNA, molecular markers of thyroid cancer can be detected as assistant method. This study aim to verify the significance of molecular detection in diagnosis of benign and malignant thyroid nodules in China. Methods: We carried out a prospective study of 383 FNA samples of thyroid nodules, and performed targeted multi-gene NGS on most samples with FSZ-Thyroid NGS Panel V1, the result of which would be incorporated into the reference index for clinicians to decide whether a patient should undergo surgery. And postoperative pathology were followed up. The NGS targets were divided into three categories: high risk (BRAF V600E, TP53, PIK3CA, AKT1, CTNNB1, RET, KRAS with VAF≥30%, HRAS with VAF≥30%, NRAS with VAF≥30%, RET fusion, NTRK1 fusion, NTRK3 fusion, ALK fusion, BRAF fusion), low risk (BRAF non-V600E, EIF1AX, GNAS, TSHR, TERT, KRAS with VAF < 30%, HRAS with VAF < 30%, HRAS with VAF < 30%, PPARG fusion, THADA fusion), benign like (EZH1, SPOP, ZNF148 or no mutations). Results: Among the 383 FNA samples, 333 of them were sequenced with FSZ-Thyroid NGS Panel V1, and 211 thyroid nodules were resected. Finally, postoperative pathology showed that 180 nodules was malignant and 31 was benign. Among the 180 malignant tumors, BRAF V600E (71.1%), RET fusion (7.8%), and TERT mutation (2.8%) were the top three most mutated genes; and there were also significant differences in frequency of some mutations between malignant tumors in different BSRTC class, including more BRAF V600E (83.6%) in class VI malignant tumors, more gene fusions (26.1%) in class V malignant tumors which was significantly higher than that in class VI (7%). And, the frequency of RAS mutations (8.3%) in class III and IV malignant tumors was higher than that of class V and VI. Here a test was considered as positive if a genetic alteration was annotated with High-Risk, and negative if Low-Risk or Benign-Like, the sensitivity and specificity for detecting malignant tumors in BSRTC class III-IV were 66.7% and 100%, respectively, and the total sensitivity and specificity for BSRTC class I-VI malignant tumors were 85.6% and 100%. In addition, 21 of 383 patients underwent Re-FNA. Among these 21 patients, 11 patients detected high risk mutations and 90.9% (10/11) underwent upgrade of BSRTC class on Re-FNA. As a comparation, among the remaining 10 patients who detected low risk or benign like sites, only 30% (3/10) underwent upgrade of BSRTC class on Re-FNA which suggested that the detection of high risk sites might predict the upgrade of BSRTC class on Re-FNA. Conclusions: Molecular testing can distinguish between benign and malignant thyroid nodules and predict the upgrade of BSRTC class on Re-FNA which could provide the reference for treatment decision-making of thyroid nodules in China.

SHMT2 expression as a diagnostic and prognostic marker for thyroid cancer.

BackgroundCatabolism of serine via serine hydroxymethyltransferase2 (SHMT2) through the mitochondrial one-carbon unit pathway is important in tumorigenesis. Therefore, SHMT2 may play a role in thyroid cancer.MethodsThyroid tissue samples and The Cancer Genome Atlas (TCGA) database were used to evaluate SHMT2 expression in thyroid tissues and the association with clinical outcomes.ResultsSHMT2 protein expression was evaluated in thyroid tissues consisting of 52 benign nodules, 129 papillary thyroid carcinomas (PTC) and matched normal samples, and 20 anaplastic thyroid carcinomas (ATC). ATCs presented the highest (95.0%) positivity of SMHT2 protein expression. PTCs showed the second highest (73.6%) positivity of SHMT2 expression, which was significantly higher than that of benign nodules (19.2%, P = 0.016) and normal thyroid tissues (0%, P < 0.001). Analysis of TCGA data showed that SHMT2 messenger RNA (mRNA) expression was significantly higher in tumors than in normal tissues (P < 0.001). When we classified thyroid cancer into high and low groups according to SHMT2 mRNA expression levels, the thyroid differentiation score for the high SHMT2 group was significantly lower than that of the low SHMT2 group (P < 0.001). There was also a significant correlation between SHMT2 mRNA expression and the stemness index (r = 0.41, P < 0.001). The high SHMT2 group had more advanced TNM stages and shorter progression-free survival rates than the low SHMT2 group (P < 0.01 and P = 0.007, respectively).ConclusionSHMT2 expression is higher in thyroid cancers than normal or benign tissues and is associated with de-differentiation and poor clinical outcomes. Thus, SHMT2 might be useful as a diagnostic and prognostic marker for thyroid cancer.

Cross validation methods: Analysis based on diagnostics of thyroid cancer metastasis

Much of modern medical research are supported by statistics and machine learning techniques. For the reliability of these studies, it is important to measure the effectiveness of the developed models in making predictions on real medical data. Cross validation is a tool, which provides a principled method of measuring the effectiveness of models and comparing models with each other. This paper presents theoretical and empirical analysis of common cross validation methods. Using sample logistic regression model along with six different validation methods, comparative analysis has been conducted. The model was developed to determine the level of thyroglobulin hormone, thyroid cancer marker. The marker indicates an increased risk of thyroid cancer metastasis, and is commonly used in metastasis diagnosis. Whereas, literature does not provide a single threshold of thyroglobulin value, indicating an increased risk of metastasis. This paper confirms that one of the main reasons of this lack, is the high variance of the developed models. This research argues that the use of the validation technique also influences both: measures of the model quality and the threshold value. The results show a high discrepancy in the determination of threshold values. However, iterative methods (such as sampling and bootstrap) seem to produce more stable outcomes.

Galectin-3 Inhibitors Suppress Anoikis Resistance and Invasive Capacity in Thyroid Cancer Cells.

Accumulating evidence suggests that galectin-3 is a histologic marker of thyroid cancer. However, the pharmacological lectin-based approach has not been well studied. In the present study, we aimed to investigate the therapeutic potential of novel galectin-3 inhibitors by treating thyroid cancer cells with different concentrations of GB1107 or TD139. At high doses, TD139, but not GB1107, reduced cell viability and clonogenicity of thyroid cancer cells. TD139 induced apoptosis of thyroid cancer cells, as evident by an increase in the percentage of sub-G1 cells on cell cycle analysis, caspase-3 activation, and PARP1 cleavage. Either GB1107 or TD139 significantly inhibited cell coherence and counteracted anoikis resistance. Both inhibitors decreased migratory and invasive abilities in a dose-dependent manner. Furthermore, GB1107 and TD139 treatment attenuated AKT phosphorylation and decreased the expression of β-catenin and MMP2. In conclusion, these novel galectin-3 inhibitors suppressed the anoikis resistance, motility, and invasive capacity of thyroid cancer cells at least partly through the AKT/β-catenin pathway. Galectin-3 inhibitors are potentially suitable for preclinical evaluation of treatment and/or prevention of metastatic spread in thyroid cancer.

NTRK Fusion Genes in Thyroid Carcinomas: Clinicopathological Characteristics and Their Impacts on Prognosis.

Simple SummaryNTRK fusion genes are important but not well studied molecular markers in thyroid cancer. Their identification could help improve diagnosis and prognosis, and determine appropriate treatment. The aims of this study were to identify NTRK fusion-positive thyroid tumors in a large cohort of different thyroid tumors, to characterize these tumors by molecular, clinical and pathological features and to evaluate the impact of NTRK-rearranged tumors on prognosis of the disease. A suitable approach for selective NTRK fusion gene testing in thyroid cancer samples was created and utilized. In a cohort of 59 NTRK-rearranged carcinomas, characteristic features were described and recommendations for surgery and prognostic factors were determined thanks to the long-term follow-up of patients.Chromosomal rearrangements of NTRK genes are oncogenic driver mutations in thyroid cancer (TC). This study aimed to identify NTRK fusion-positive thyroid tumors and to correlate them with clinical and pathological data and determine their prognostic significance. The cohort consisted of 989 different TC samples. Based on the detected mutation, samples were triaged, and those that were positive for a BRAF, HRAS, KRAS, NRAS, RET, RET/PTC or PAX8/PPARγ mutation were excluded from further analyses. NTRK fusion gene testing was performed in 259 cases, including 126 cases using next-generation sequencing. NTRK fusion genes were detected in 57 of 846 (6.7%) papillary thyroid carcinomas and in 2 of 10 (20.0%) poorly differentiated thyroid carcinomas. A total of eight types of NTRK fusions were found, including ETV6/NTRK3, EML4/NTRK3, RBPMS/NTRK3, SQSTM1/NTRK3, TPM3/NTRK1, IRF2BP2/NTRK1, SQSTM1/NTRK1 and TPR/NTRK1. NTRK fusion-positive carcinomas were associated with the follicular growth pattern, chronic lymphocytic thyroiditis and lymph node metastases. NTRK1-rearranged carcinomas showed a higher frequency of multifocality and aggressivity than NTRK3-rearranged carcinomas. Tumor size, presence of metastases, positivity for the NTRK3 or NTRK1 fusion gene and a late mutation event (TERT or TP53 mutation) were determined as factors affecting patient prognosis. NTRK fusion genes are valuable diagnostic and prognostic markers.

Immunohistochemical Expression of Galectin-3 and Cytokeratin 19 in the Spectrum of Thyroid Neoplasms

Introduction: In systemic malignancies, thyroid carcinoma represents only 1%, but it is the most common endocrine malignancy with poor prognosis. Fine Needle Aspiration (FNA) is considered a requisite tool in providing a rational advent for the clinical management of these nodules. However, when solely based on cytopathological assessment, FNA leads to imprecise biopsy results in 10-20% of all cases. So, immunohistochemical markers, Galectin-3 (Gal-3) and Cytokeratin 19 (CK19) have received considerable attention as diagnostic marker for thyroid cancer. Aim: To study the expression of Gal-3 and CK19 in different thyroid neoplasms. Materials and Methods: This cross-sectional study was conducted at a tertiary care hospital from January 2019 to January 2021 on 120 cases. The thyroidectomy specimens received were fixed in 10% buffered formalin followed by gross examination. The paraffin embedded tissue blocks were subsequently stained for Haematoxylin and Eosin stain (H&amp;E) followed by histopathological reporting. Qualitative immunohistochemical assessment of the marker Gal-3 and CK19 was performed on representative histologic sections of the thyroid neoplasms. Data were analysed using Graph Pad Instat 3. Results: The positive expression of Gal-3 was significantly more in malignant tumours (87.5%) than in benign (36.0%) (p-value &lt;0.001). In Papillary Thyroid Carcinomas (PTC) and in Follicular Variant of Papillary Thyroid Carcinomas (FVPTC) expression of Gal-3 was notably higher than in Follicular Adenoma (FA) (p-value=0.01 and p-value=0.0001, respectively). Follicular Thyroid Carcinomas (FTC) had higher expression FA (p-value=0.003). In malignant tumours (71.8%), positive expression of CK19 was significantly more than benign tumours (24.0%) (p-value &lt;0.001). Significant difference in expression of CK19 was seen between PTC and FTC (p-value=0.019). Between PTC and FA, differences in expression were significant (p-value &lt;0.001). Significant difference in expression was also seen between FVPTC and FTC (p-value=0.032) as well as with FA (p-value=0.028). Conclusion: The most sensitive marker is Gal-3 for the diagnosis of thyroid malignancies. When combined with CK19, the specificity increases in identifying the thyroid cancers. More combination of markers together with Gal-3 and CK19 can be useful in the distinction between malignant and benign thyroid tumours because it is essential for further treatment and long-term management of the patient.

Clinicopathologic Analysis of Cathepsin B as a Prognostic Marker of Thyroid Cancer.

Thyroid cancer incidence has increased worldwide; however, investigations of thyroid cancer-related factors as potential prognosis markers remain insufficient. Secreted proteins from the cancer secretome are regulators of several molecular mechanisms and are, thereby, ideal candidates for potential markers. We aimed to identify a specific factor for thyroid cancer by analyzing the secretome from normal thyroid cells, papillary thyroid cancer (PTC) cells, and anaplastic thyroid cancer cells using mass spectrometry (MS). Cathepsin B (CTSB) showed highest expression in PTC cells compared to other cell lines, and CTSB levels in tumor samples were higher than that seen in normal tissue. Further, among thyroid cancer patients, increased CTSB expression was related to higher risk of lymph node metastasis (LNM) and advanced N stage. Overexpression of CTSB in thyroid cancer cell lines activated cell migration by increasing the expression of vimentin and Snail, while its siRNA-mediated silencing inhibited cell migration by decreasing vimentin and Snail expression. Mechanistically, CTSB-associated enhanced cell migration and upregulation of vimentin and Snail occurred via increased phosphorylation of p38. As our results suggest that elevated CTSB in thyroid cancer induces the expression of metastatic proteins and thereby leads to LNM, CTSB may be a good and clinically relevant prognostic marker.

Mechanisms of TERT Reactivation and Its Interaction with BRAFV600E.

The telomerase reverse transcriptase (TERT) gene, which is repressed in most differentiated human cells, can be reactivated by somatic TERT alterations and epigenetic modulations. Moreover, the recruitment, accessibility, and binding of transcription factors also affect the regulation of TERT expression. Reactivated TERT contributes to the development and progression of cancer through telomere lengthening-dependent and independent ways. In particular, because of recent advances in high-throughput sequencing technologies, studies on genomic alterations in various cancers that cause increased TERT transcriptional activity have been actively conducted. TERT reactivation has been reported to be associated with poor prognosis in several cancers, and TERT promoter mutations are among the most potent prognostic markers in thyroid cancer. In particular, when a TERT promoter mutation coexists with the BRAFV600E mutation, these mutations exert synergistic effects on a poor prognosis. Efforts have been made to uncover the mechanisms of these synergistic interactions. In this review, we discuss the role of TERT reactivation in tumorigenesis, the mechanisms of TERT reactivation across all human cancers and in thyroid cancer, and the mechanisms of interactions between BRAFV600E and TERT promoter mutations.

157P Search for new markers of thyroid carcinogenesis associated with ER, AR and PR receptor status

In recent years a plethora of new investigating methods have appeared, which allows us to fully and thoroughly observe thyroid diseases. Though, final diagnosis at the pre-operative stage remains to be difficult. Fine-needle aspiration biopsy used for pre-surgical diagnosis doesn't distinguish benign follicular tumors (adenoma) from malignant ones (carcinoma). It is known that thyroid cancer is more common in women (approximately 1: 3), but more aggressive forms of thyroid cancer are common in men. This indicates that estrogen, progesterone and androgen receptors (ER, PR and AR) can play an important role in carcinogenesis of the thyroid gland. Also miRNAs, which role in thyroid cancerogenesis processes is being actively studied, are in interest. However, certain attempts to connect disruptions in receptor expression with disruptions in miRNA expression during thyroid cancerogenesis haven’t been made yet. The expression of the miRNAs and their target-genes was analyzed by RT-PCR in 90 samples of thyroid neoplasms (TN). To find new thyroid cancer markers, we analyzed the expression levels of ER, PR, AR, miRNAs for which these receptors are targets, as well as other target genes of selected miRNAs (ID4, FOXE1, VEGFA, BCL2, ITGB3, SLC24A4, TSHR) in TN. The results has shown that benign TN can be distinguished from malignant ones by expression levels of miRNA-181a, miRNA-205 and their target genes – PR, VEGFA, BCL2, TSHR. Markers, that allow to distinguish follicular variant of papillary thyroid cancer from follicular and papillary carcinomas, have been identified as well. Moreover, it has been shown that miRNA-22 can be marker for lymph node metastases: miRNA level is reduced in patients with metastases. Furthermore, based on the results, it can be assumed that there is a relationship between miRNA-222, miRNA-19b, miRNA-22, miRNA-193b, miRNA-205, miRNA-181a, miRNA-378a levels and ER, PR, AR expression levels during thyroid tumor development. Markers that distinguish between different types of thyroid tumors have been identified (miRNA-181a, miRNA-205, PR, VEGFA, BCL2 and TSHR). The miRNA-22 level was shown to be associated with N stage.

The Potential of Metabolomics in the Diagnosis of Thyroid Cancer.

Thyroid cancer is the most common endocrine system malignancy. However, there is still a lack of reliable and specific markers for the detection and staging of this disease. Fine needle aspiration biopsy is the current gold standard for diagnosis of thyroid cancer, but drawbacks to this technique include indeterminate results or an inability to discriminate different carcinomas, thereby requiring additional surgical procedures to obtain a final diagnosis. It is, therefore, necessary to seek more reliable markers to complement and improve current methods. “Omics” approaches have gained much attention in the last decade in the field of biomarker discovery for diagnostic and prognostic characterisation of various pathophysiological conditions. Metabolomics, in particular, has the potential to identify molecular markers of thyroid cancer and identify novel metabolic profiles of the disease, which can, in turn, help in the classification of pathological conditions and lead to a more personalised therapy, assisting in the diagnosis and in the prediction of cancer behaviour. This review considers the current results in thyroid cancer biomarker research with a focus on metabolomics.

Linc00941 Is a Novel Transforming Growth Factor β Target That Primes Papillary Thyroid Cancer Metastatic Behavior by Regulating the Expression of Cadherin 6.

Background: Papillary thyroid cancers (PTCs) are common, usually indolent malignancies. Still, a small but significant percentage of patients have aggressive tumors and develop distant metastases leading to death. Currently, it is not possible to discriminate aggressive lesions due to lack of prognostic markers. Long noncoding RNAs (lncRNAs), which are selectively expressed in a context-dependent manner, are expected to represent a new landscape to search for molecular discriminants. Transforming growth factor β (TGFβ) is a multifunctional cytokine that fosters epithelial-to-mesenchymal transition and metastatic spreading. In PTCs, it triggers the expression of the metastatic marker Cadherin 6 (CDH6). Here, we investigated the TGFβ-dependent lncRNAs that may cooperate to potentiate PTC aggressiveness. Methods: We used a genome-wide approach to map enhancer (ENH)-associated lncRNAs under TGFβ control. Linc00941 was selected and validated using functional in vitro assays. A combined approach using bioinformatic analyses of the thyroid cancer (THCA)-the cancer genome atlas (TCGA) dataset and RNA-seq analysis was used to identify the processes in which linc00941 was involved in and the genes under its regulation. Correlation with clinical data was performed to evaluate the potential of this lncRNA and its targets as prognostic markers in THCA. Results: Linc00941 was identified as transcribed starting from one of the TGFβ-induced ENHs. Linc00941 expression was significantly higher in aggressive cancer both in the TCGA dataset and in a separate validation cohort from our institution. Loss of function assays for linc00941 showed that it promotes response to stimuli and invasiveness while restraining proliferation in PTC cells, a typical phenotype of metastatic cells. From the integration of TCGA data and linc00941 knockdown RNA-seq profiling, we identified 77 genes under the regulation of this lncRNA. Among these, we found the prometastatic gene CDH6. Linc00941 knockdown partially recapitulates the effects observed upon CDH6 silencing, promoting cell cytoskeleton and membrane adhesions rearrangements and autophagy. The combined expression of CDH6 and linc00941 is a distinctive feature of highly aggressive PTC lesions. Conclusions: Our data provide new insights into the biology driving metastasis in PTCs and highlight how lncRNAs cooperate with coding transcripts to sustain these processes.

The Blood Biomarkers of Thyroid Cancer.

IntroductionWith the gradual increase in the incidence of thyroid cancer, people’s attention to thyroid cancer has also gradually increased. Although the prognosis of thyroid cancer is rather mild compared to other cancers, it will still bring a heavy psychological burden on people who have been diagnosed. At present, the diagnosis of thyroid cancer mainly depends on ultrasound and percutaneous fine needle aspiration (pFNA). Due to the unsatisfactory accuracy of the diagnosis methods we use now, there are still some thyroid nodules that cannot be clearly diagnosed before surgery.MethodsIn this article, we have searched for relevant research on blood markers of thyroid cancer in the past five years and categoried them into four groups.DiscussionThough we have not found a biomarker which can diagnose thyroid cancer both sensitively and specifically, we do found many substances that are related to it, and have the potential to recognize it and help the diagnosis. And perhaps combined models can do it better.

Significance of Insulin-Like Growth Factor II Mrna-Binding Protein 3 (IMP3) Expression in Selected Thyroid Lesions

Background: Differentiation of benign from malignant follicular thyroid lesions remains difficult and the ability of molecular markers to differentiate between them still unclear. The aim of this study was to evaluate the usefulness of IMP3 expression to distinguish benign from malignant thyroid lesions. Methods: This is a retrospective study upon selected 57 thyroid lesions designated as; 7 cases of Hashimoto thyroiditis (HT), 10 cases of hyperplastic nodules (HN), 15 cases of follicular thyroid adenoma (FTA), 13 cases of conventional papillary thyroid carcinoma (PTC), 6 cases of follicular variant of papillary thyroid carcinoma (FV-PTC), and 6 cases of follicular thyroid carcinoma (FTC). Immunohistochemistry was applied on formalin-fixed, paraffin-embeded tissue blocks using IMP3. clinicopathological data were reviewed from patients' pathological reports and correlated with IMP3 expression. Results: IMP3 positivity was seen in1/7 cases (14.3%) of HT, 2/10 cases (20%) of HN, 4/15 cases (26.7%) of FTA, 12/13 cases of PTC, and in all (100%) FV-PTC & FTC cases. IMP3 staining was significantly increased from normal thyroid tissue up to malignant tumors (P<0.01). IMP3 showed 96% sensitivity and 78.1% specificity for malignant tumors. IMP3 expression was positively correlated with grade and tumor size in malignant cases (P<0.05). No significant correlation was found in IMP3 expression with patient age, sex, capsular invasion, lymphatic/vascular invasion, lymph node metastasis, distant metastasis and TNM stage. Conclusions: IMP3 is a potential diagnostic marker for thyroid cancer and can be a promising marker for distinguishing benign from malignant follicular patterned thyroid lesions.

Highly accurate NGS-based multi-gene testing in the diagnosis of thyroid nodules with indeterminate cytology.

e13579 Background: Thyroid nodules are highly prevalent, Fine-needle aspiration (FNA) is the standard pre-operative tool for diagnosis. However, some of the samples are classified as indeterminate, which leads to unnecessary surgery. BRAF V600E mutation is often used as a diagnostic marker for thyroid cancer, and it is highly specific for papillary thyroid carcinoma (PTC). But BRAF mutation is rarely occurred in thyroid nodules with indeterminate cytology. To diagnose the indeterminate thyroid nodules precisely, some NGS-based multi-gene testing panel has been developed and clinically used in America and Europe, but rare research was reported in China. In this study, we evaluated the value of a next-generation sequencing (NGS) panel to cancer diagnosis in indeterminate thyroid nodules. Methods: From February 2018 to September 2018, 360 patients with thyroid nodules who underwent FNA at Peking University Cancer Hospital were enrolled. And the FNA samples with indeterminate cytology were evaluated using a next-generation sequencing (NGS) assay, including 16 genes analyzed for point mutations and 26 types of gene fusions. Diagnostic performance of this multi-gene testing panel was compared with BRAF V600E single gene mutation analysis. Results: 141 nodules were cytologically indeterminate among 360 patients on FNA biopsy, 72 of which were resected and analyzed by NGS successfully. Histologic analysis after surgery revealed 41 (56.9%) cancers in these 72 patients. The multi-gene testing assay could classify 30/41 cancers correctly, showing a sensitivity of 73.2%, specificity of 96.8%, positive predictive value of 96.8%, and negative predictive value of 73.2%. The diagnostic accuracy of the multi-gene testing was significantly higher than the BRAF V600E mutation analysis (83.3% vs 73.6%, x2= 31.588, p &lt; 0.01). Conclusions: Our study demonstrated that the multi-gene testing provided both high sensitivity and high specificity for cancer detection in thyroid nodules with indeterminate cytology, and its accuracy was much higher than BRAF V600E mutation test.

MON-LB75 Stratifying the Risk of Malignancy in Indeterminate Thyroid Nodules

Background Thyroid cancer is the most common endocrine cancer globally, and accounts for 3.4% of all new cancer cases in the US. Although several clinical guidelines to detect/manage thyroid nodules are available, a great deal of controversy still exists around the optimal approach for diagnosis. Approximately 20-30% of cytology results from thyroid fine-needle aspiration (FNA) fall into one of three indeterminate diagnostic categories. In recent years, a number of molecular and gene mutation diagnostic tests have been developed to diagnose the indeterminate thyroid nodules in FNA specimens. However, nearly half of patients recommended for surgery based on these tests were found to have a benign nodule. Therefore, there is a need for a more accurate predictive and prognostic test for thyroid cancer. In case of Euthyroid Hashimoto Thyroiditis condition (EHT), where patient required a partial thyroxin replacement dose, having about 48% risk of thyroid cancer. However, there is still not a very accurate predictive marker for early detection of thyroid cancer in EHT. Invention DescriptionResearchers at the University of Toledo have found that thyroid nodule microenvironment cell profiling can be used as a predictive and prognostic marker for thyroid cancer. This approach uniquely focuses on the phenotype, rather than the genotype, of the microenvironment. Recently described Double Negative (DN) T-cells were significantly more abundant in lymphocytic infiltrates of thyroid cancer. They were shown to downregulate proliferation and cytokine production of activated effector T cells present in the tumor microenvironment and contribute to tumor tolerance and active avoidance of tumor immunity. If the quantify of DN T cells exceeds the defined threshold, it indicates a likelihood of cancer presence. Aside from immune cell profiling, our approach further establishes an integration of the information derived from transcriptome/meta-analysis of the genome and cytokine/chemokine signal analysis all from thyroid FNA. Applications[[Unsupported Character - Symbol Font •]] Diagnosis of thyroid cancer from FNA samples[[Unsupported Character - Symbol Font •]] Predictive tool of severity of disease Advantages[[Unsupported Character - Symbol Font •]] Microenvironment profiling can provide a unique way to diagnose and assess disease progression[[Unsupported Character - Symbol Font •]] Sheds light on cellular cross-talk; more accurate diagnose; can prevent unnecessary surgery IP Status:​Patent Pending

Hsa_circ_0124055 and hsa_circ_0101622 regulate proliferation and apoptosis in thyroid cancer and serve as prognostic and diagnostic indicators.

Recent studies have corroborated that circular RNAs (circRNAs) as endogenous noncoding RNAs gain research interest in carcinogenesis, functioning as prognostic and diagnostic biomarkers and therapeutic targets. The present study is aimed to determine whether circRNAs could serve as prognostic and diagnostic biomarkers to predict thyroid carcinoma. High-throughput sequencing analysis was conducted to detect circRNAs expression profile in thyroid cancer. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) measurement was utilized to validate circRNAs expression in blood and tissue specimens. Kaplan-Meier method and receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC) were used to assess whether circRNAs could function as prognostic and diagnostic biomarkers of thyroid cancer, respectively. Hsa_circ_0124055 and hsa_circ_0101622 as the most conspicuous biomarkers were significantly increased in tumor tissues and plasma of thyroid cancer patients. High hsa_circ_0124055 or hsa_circ_0101622 expression exhibited shorter overall survival. Our findings also provided strong evidence that plasma hsa_circ_0124055 (AUC = 0.836, 95% CI: 0.763-0.908, p<0.001) and hsa_circ_0101622 (AUC = 0.805, 95% CI: 0.727-0.883, p<0.001) could be used as diagnostic markers for thyroid cancer, and hsa_circ_0124055 combined with hsa_circ_0101622 could provide a more powerful diagnostic value (AUC = 0.911, 95% CI: 0.859-0.962, p<0.001) than the use of hsa_circ_0124055 or hsa_circ_0101622 alone. Furthermore, the knockdown of hsa_circ_0124055 or hsa_circ_0101622 exhibited a significant anti-proliferative and pro-apoptotic activity of thyroid cancer cells in vivo and in vitro. Both hsa_circ_0124055 and hsa_circ_0101622 could facilitate the prognosis and diagnosis of thyroid cancer, and function as the therapeutic targets for clinical practice.

Identification of Potential Biomarkers for Thyroid Cancer Using Bioinformatics Strategy: A Study Based on GEO Datasets.

Background The molecular mechanisms and genetic markers of thyroid cancer are unclear. In this study, we used bioinformatics to screen for key genes and pathways associated with thyroid cancer development and to reveal its potential molecular mechanisms. Methods The GSE3467, GSE3678, GSE33630, and GSE53157 expression profiles downloaded from the Gene Expression Omnibus database (GEO) contained a total of 164 tissue samples (64 normal thyroid tissue samples and 100 thyroid cancer samples). The four datasets were integrated and analyzed by the RobustRankAggreg (RRA) method to obtain differentially expressed genes (DEGs). Using these DEGs, we performed gene ontology (GO) functional annotation, pathway analysis, protein-protein interaction (PPI) analysis and survival analysis. Then, CMap was used to identify the candidate small molecules that might reverse thyroid cancer gene expression. Results By integrating the four datasets, 330 DEGs, including 154 upregulated and 176 downregulated genes, were identified. GO analysis showed that the upregulated genes were mainly involved in extracellular region, extracellular exosome, and heparin binding. The downregulated genes were mainly concentrated in thyroid hormone generation and proteinaceous extracellular matrix. Pathway analysis showed that the upregulated DEGs were mainly attached to ECM-receptor interaction, p53 signaling pathway, and TGF-beta signaling pathway. Downregulation of DEGs was mainly involved in tyrosine metabolism, mineral absorption, and thyroxine biosynthesis. Among the top 30 hub genes obtained in PPI network, the expression levels of FN1, NMU, CHRDL1, GNAI1, ITGA2, GNA14 and AVPR1A were associated with the prognosis of thyroid cancer. Finally, four small molecules that could reverse the gene expression induced by thyroid cancer, namely ikarugamycin, adrenosterone, hexamethonium bromide and clofazimine, were obtained in the CMap database. Conclusion The identification of the key genes and pathways enhances the understanding of the molecular mechanisms for thyroid cancer. In addition, these key genes may be potential therapeutic targets and biomarkers for the treatment of thyroid cancer.

PD-L1 expression in papillary thyroid cancer with and without lymphocytic thyroiditis: a cross sectional study

The impact of concurrent autoimmune thyroid disease on the tumour microenvironment and disease progression in papillary thyroid cancer (PTC) is not well understood. Studies evaluating the programmed cell death ligand 1 (PD-L1) tumour expression in PTC have shown variable results, and the effect of lymphocytic thyroiditis (LT) on tumour PD-L1 expression has not been adequately assessed. The main aim of this study was to determine expression of PD-L1 in PTC with and without LT. We examined 81 PTC cases; 28.5% of all reviewed PTC had presence of LT. In PTC specimens without LT, tumour PD-L1 expression was significantly lower compared to PD-L1 expression in PTC with LT, 6.9% vs 39.1%, respectively. Expression of PD-L1 did not differ with PTC stage, even when sub-categorised according to the presence and absence of LT. Utility of PD- L1 expression as a prognostic marker in thyroid cancer needs to be interpreted with caution.

Hsa-miR-139-5p is a prognostic thyroid cancer marker involved in HNRNPF-mediated alternative splicing.

It is critical to identify biomarkers and functional networks associated with aggressive thyroid cancer to anticipate disease progression and facilitate personalized patient management. We performed miRNome sequencing of 46 thyroid tumors enriched with advanced disease patients with a median follow-up of 96 months. MiRNome profiles correlated with tumor-specific histopathological and molecular features, such as stromal cell infiltration and tumor driver mutation. Differential expression analysis revealed a consistent hsa-miR-139-5p downexpression in primary carcinomas from patients with recurrent/metastatic disease compared to disease-free patients, sustained in paired local metastases and validated in publicly available thyroid cancer series. Exogenous expression of hsa-miR-139-5p significantly reduced migration and proliferation of anaplastic thyroid cancer cells. Proteomic analysis indicated RICTOR, SMAD2/3 and HNRNPF as putative hsa-miR-139-5p targets in our cell system. Abundance of HNRNPF mRNA, encoding an alternative splicing factor involved in cryptic exon inclusion/exclusion, inversely correlated with hsa-miR-139-5p expression in human tumors. RNA sequencing analysis revealed 174 splicing events differentially regulated upon HNRNPF repression in our cell system, affecting genes involved in RTK/RAS/MAPK and PI3K/AKT/MTOR signaling cascades among others. These results point at the hsa-miR-139-5p/HNRNPF axis as a novel regulatory mechanism associated with the modulation of major thyroid cancer signaling pathways and tumor virulence.

Молекулярно-генетичні механізми патогенезу злоякісних пухлин щитоподібної залози (огляд літератури та власні дані, частина 1)

У 1-й частині огляду проаналізовано нові дані наукової літератури щодо молекулярно-генетичних механізмів утворення папілярної та фолікулярної карцином щитоподібної залози. Відзначено роль мутацій і перебудов генів, продукти яких формують МАРК- і РІ3К-сигнальні каскади. Аналізуються нещодавно виявлені за допомогою сучасних генетичних методів чинники, що впливають на патогенез папілярної карциноми щитоподібної залози. Особливу увагу приділено доказам впливу іонізуючої радіації на канцерогенез. Обговорюється значущість нових діагностичних і прогностичних маркерів раку щитоподібної залози.