Molecular Markers 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.

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.

Differences in miRNA expression profiles between wild-type and mutated NIFTPs.

Noninvasive encapsulated follicular variants of papillary thyroid carcinomas have been recently reclassified as noninvasive follicular thyroid neoplasms with papillary-like nuclear features (NIFTPs). NIFTPs exhibit a behavior that is very close to that of follicular adenomas but different from the infiltrative and invasive follicular variants of papillary thyroid carcinomas (FVPTCs). The importance of miRNAs to carcinogenesis has been reported in recent years. miRNAs seem to be promising diagnostic and prognostic molecular markers for thyroid cancer, and the combination of miRNA expression and mutational status might improve cytological diagnosis. The aim of the present study was to evaluate the miRNA expression profile in wild-type, RAS- or BRAF-mutated NIFTPs, infiltrative and invasive FVPTCs, and follicular adenomas using the nCounter miRNA Expression assay (NanoString Technologies). To identify the significant Kyoto Encyclopedia of Genes and Genomes (KEGG) molecular pathways associated with deregulated miRNAs, we used the union of pathways option in DNA Intelligent Analysis (DIANA) miRPath software. We have shown that the miRNA expression profiles of wild-type and mutated NIFTPs could be different. The expression profile of wild-type NIFTPs seems comparable to that of follicular adenomas, whereas mutated NIFTPs have an expression profile similar to that of infiltrative and invasive FVPTCs. The upregulation of 4 miRNAs (miR-221-5p, miR-221-3p, miR-222-3p, miR-146b-5p) and the downregulation of 8 miRNAs (miR-181a-3p, miR-28-5p, miR-363-3p, miR-342-3p, miR-1285-5p, miR-152-3p, miR-25-3p, miR-30e-3) in mutated NIFTPs compared to wild-type ones suggest a potential invasive-like phenotype by deregulating the specific pathways involved in cell adhesion and cell migration (Hippo signaling pathway, ECM-receptor interaction, adherens junction, regulation of actin cytoskeleton, fatty acid biosynthesis and metabolism).

Role of Molecular Markers in Thyroid Nodule Management: Then and Now

To describe the evolution and clinical utility of molecular testing for thyroid nodules and cancer achieved over the last 2 decades. Scientific reports on thyroid cancer genetics and molecular diagnostics in thyroid nodules. Over the last 2 decades, our understanding of the genetic mechanisms of thyroid cancer has dramatically expanded, such that most thyroid cancers now have known gene driver events. This knowledge provides the basis for establishing and further improving molecular tests for thyroid nodules and cancer and for the introduction of new entities such as noninvasive follicular thyroid neoplasm with papillary-like nuclear features. The progress with molecular tests for thyroid nodules started in the 1990s from demonstrating feasibility of detecting various molecular alterations in fine-needle aspiration (FNA) material collected from thyroid nodules. It was followed by the introduction of the first single-gene mutational markers, such as BRAF, and a small mutational panel into clinical practice in the mid 2000s. Currently, several more advanced molecular tests are available for clinical use. They are based on multiple molecular markers and have increasing impact on the clinical management of patients with thyroid nodules. The evolution of molecular tests for thyroid nodules followed the discovery of various diagnostic and prognostic molecular markers of thyroid cancer that can be applied to thyroid FNA samples to inform more individualized management of these patients. FNA = fine-needle aspiration miRNA = micro RNA NGS = next-generation sequencing NIFTP = noninvasive follicular thyroid neoplasm with papillary-like nuclear features NPV = negative predictive value PPV = positive predictive value PTC = papillary thyroid carcinoma RAI = radioactive iodine.

Expression, clinical significance and mechanism of Slit2 in papillary thyroid cancer

Thyroid cancer is a common endocrine malignancy. The last decade has seen exciting progress in understanding thyroid cancer molecular pathogenesis. Several major signaling pathways and related molecular derangements have been elucidated, which represent novel diagnostic and prognostic molecular markers for thyroid cancer. Based on the molecular biology of thyroid cancer, a series of therapeutic targets have been developed, which provide unprecedented opportunities. Thus, histological characterization of subgroups of patients and the correct molecular characterization of patients are thought to be key aspects for future clinical management of these patients. In the present study, we identified Slit2 as a prognostic marker for thyroid cancer oncogenesis and recurrence. Mechanistically, Slit2 regulated Warburg effect in thyroid cancer cells through regulation of HIF1α and HIF1α transcriptional activity. Taken together, our present data uncovered Slit2 as a novel predictive marker for thyroid cancer. The mechanism study indicated that Slit2 regulated the Warburg effect. Additional study on the function of Slit2 in thyroid cancer is required to provide new insights into the potential mechanisms of oncogenesis and recurrence potential of thyroid cancer.

Protein molecular modeling of genetic markers for thyroid cancer

Introduction: The advances in thyroid molecular biology studies provide not only insight into thyroid diseases but accurate diagnosis of thyroid cancer. Objective: Design a tutorial on protein molecular modeling of genetic markers for thyroid cancer. Methods: The proteins were selected using the Protein Data Bank sequence and the basic local alignment search tool (BLAST) algorithm. The obtained sequences were aligned with the Clustal W multiple alignment algorithms. For the molecular modeling, three-dimensional structures were generated from this set of constraints with the SWISS-MODEL, which is a fully automated protein structure homology-modeling server, accessible via the ExPASy web server. Results: We demonstrated protein analysis, projection of the molecular structure and protein homology of the following molecular markers of thyroid cancer: receptor tyrosine kinase (RET) proto-oncogene; neurotrophic tyrosine kinase receptor 1 (NTRK1) proto-oncogene; phosphatase and tensin homolog (PTEN); tumor protein p53 (TP53) gene; phosphoinositide 3-kinase/threonine protein kinase (PI3K/AKT); catenin beta 1 (CTNNB1); paired box 8-peroxisome proliferator-activated receptor gamma (PAX8-PPARG); rat sarcoma viral oncogene (RAS); B-raf proto-oncogene, serine/threonine kinase (BRAF); and thyroid-stimulating hormone receptor (TSHR). Conclusion: This study shows the importance of understanding the molecular structure of the markers for thyroid cancer through bioinformatics, and consequently, the development of more effective new molecules as alternative tools for thyroid cancer treatment.

Advances and practical use of the molecular markers for thyroid cancer

Background : Thyroid nodules are very common; however, only 5–15% of nodules are malignant. Although most of the malignant thyroid cancers can be identified pre-operatively by cytological analysis, approximately 20–30% of these nodules are classified indeterminate. Subsequently, repeated Fine Needle Aspiration (FNA) or thyroid surgery may be required for a more definitive diagnosis. In the United States, the incidence of thyroid cancer has tripled from 1975 with an incidence rate of 14.3 per 100,000 individuals, primarily due to an increased diagnosis of papillary thyroid cancer which increased by 9.1 per 100,000 (RR 3.7, 95% CI (3.4–4.0)); however, mortality has remained stable. Many patients with cytologically indeterminate nodules undergo unnecessary diagnostic surgeries, placing them at risk of potential surgical complications and need for lifelong levothyroxine replacement. The advent of molecular gene testing has drawn significant attention toward improved stratification of these indeterminate lesions into benign or malignant. Detection of any pertinant molecular mutations in the indeterminate categories- atypia of undetermined significance (AUS)/ follicular lesion of undetermined significance (FLUS), follicular neoplasm (FN)/suspicious for a follicular neoplasm (SFN), and suspicious for malignant cells (SMC) has been shown to confer the risk of histologic malignancy of 88, 87 and 95% respectively, while a negative test for the mutation was reassuring with a very high negative predictive value over 95%. Purpose : The purpose of this article is to review the current recommendations to implement molecular genetic testing for thyroid nodules. We review the most common gene mutations harbored in the various thyroid cancers, including BRAF, RAS, and RET/PTC. We will also review the more recently discovered gene mutations for thyroid cancer, including TERT mutations. Finally, we will discuss the practice guidelines to implement molecular gene testing for the indeterminate cytology of thyroid nodules as well as the new category of non-invasive follicular thyroid neoplasm with papillary-like nuclear features as an intermediate pathology which will not require aggressive therapy after diagnostic lobectomy. (Published: 13 December 2016) Citation: Advances in Cellular and Molecular Otolaryngology 2016, 4: 33948 - http://dx.doi.org/10.3402/acmo.v4.33948

Molecular approaches to thyroid cancer diagnosis.

Thyroid nodules are common, and the accurate diagnosis of cancer or benign disease is important for the effective clinical management of patients. Molecular markers are a helpful diagnostic tool, particularly for cytologically indeterminate thyroid nodules. In the past few years, significant progress has been made in developing molecular markers for clinical use in fine-needle aspiration specimens, including gene mutation panels and gene expression classifiers. With the availability of next generation sequencing technology, gene mutation panels can be expanded to interrogate multiple genes simultaneously and to provide yet more accurate diagnostic information. In addition, recently several new molecular markers of thyroid cancer have been identified that offer diagnostic, prognostic, and therapeutic information that might be of value in guiding individualized management of patients with thyroid nodules.

BRAF V600 co‐testing is technically feasible in conventional thyroid fine needle aspiration (FNA) cytology smears and can reduce the need for completion thyroidectomy

While fine needle aspiration cytology (FNAC) is the mainstay of diagnosis in thyroid nodules, molecular markers of thyroid cancer have recently been shown to be of value in improving the diagnosis and reducing the rates of unnecessary surgery. A technical method is presented for the assessment of the BRAF V600 gene mutation in thyroid cancer using a simple adaptation of a commercially available kit. After standard preparation and reporting of conventionally stained alcohol-fixed Papanicolaou or air-dried Giemsa-stained slides the coverslip is removed from one slide, the DNA is extracted and submitted for PCR analysis. Assessment of the BRAF V600 mutational status is feasible in very small quantities of DNA, requiring just greater than 5ng per case from a single pre-stained FNA slide using this method. From the 14 cases examined thus far, one Thy4/Bethesda Class V case (suspicious of malignancy) has been identified with a BRAF V600 mutation and this patient, after multidisciplinary discussion, received a total thyroidectomy. Based on this methodology and other published results for the BRAF mutation, we believe that it is now feasible and cost effective for the UK NHS to BRAF co-test all Thy4/Bethesda Class V thyroid FNAs, as the additional cost of BRAF testing will still be much less than the cost of submitting all Thy4 (Bethesda Class V) patients to a partial and then a later completion thyroidectomy.

Role of Galectin-3 in Thyroid Cancer

Copyright: © 2012 Htwe TT. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Thyroid cancer (TC) is the most common endocrine malignancy with a rising incidence. It has a wide spectrum of clinical behavior and varied therapeutic responsiveness. Among the various forms of TC, papillary thyroid cancer (PTC) accounts for 85% of all TCs. It is the most common form compared to follicular carcinoma (FCA) and other rare forms such as medullary carcinoma (MCA), anaplastic carcinoma (ANA), poorly differentiated carcinoma (PDTC), undifferentiated thyroid carcinoma (UDTC), Hurthle cell carcinoma (HCA) and Insular carcinoma (Ins). In the past, PTC and FCA were generally identified by the difference in their clinical nature and the characteristic morphological features by using the conventional way of microscopy, following the guidelines from the World Health Organization (WHO). PTC is identified by its peculiar nuclear features and the papillary pattern whereas in FCA it is from its follicular pattern. The main criterion for the diagnosis of PTC is the occurrence of enlarged, round to oval shaped overlapping ground-glass nuclei, well known as OrphanAnnie eyes. The delicate papillary pattern of growth and the presences of laminated calcifications (psammoma bodies) are also diagnostic. For the FCA it is based on its closely packed follicular pattern and its tendency in invasiveness. However, there is a difficulty in identifying one sub type of TC known as papillary thyroid carcinoma follicular variant (PTCFC). Henceforth, with the development in immunohistochemical (IHC) techniques, experiments have been performed to find out novel molecular markers for TC, where Galectin-3 (Gal-3) has been identified as the most useful tumor marker among other galactosidebinding proteins such as Gal-1 and Gal-7.

Review of Molecular Markers for Thyroid Cancer

The incidence of thyroid cancer has been rapidly increased in Korea. Although fine needle aspiration cytology is recommended for diagnosis of cancer, there are some limitations. Patients with indeterminate or suspicious cytology category in which malignancy cannot be ruled out usually undergone a thyroidectomy, however, only 10∼25% of them finally diagnosed as cancer. According to the progress in understanding molecular mechanism, some mutations or other molecular alterations have been studied for the diagnostic and prognostic markers for thyroid cancer. The majority of papillary thyroid cancers have BRAF and RAS mutations or RET/PTC rearrangement, and approximately 80% of follicular thyroid cancers harbor a RAS mutation or PAX8/PPARγ rearrangement. These genetic alterations are mostly studied and current clinical guidelines suggested that these molecular markers may help management for patients with indeterminate cytology. In addition, recent studies demonstrated the high sensitivity and specificity of thyroid-stimulating hormone receptor mRNA in diagnosing cancer in patients with indeterminate cytology. For the detection of recurrent or residual thyroid cancer, serum thyroglobulin is the only circulating marker in clinical practice. However, it lacks sensitivity and is unreliable specifically in the presence of antibodies to thyroglobulin. Recent studies demonstrated a significant role of measuring the mRNA of thyroglobulin, thyroid peroxidase, thyroid-stimulating hormone receptor, and sodium/iodine symporter in peripheral blood for monitoring of the recurrence of thyroid cancer. (Ewha Med J 2012;35(1):3-10)

Molecular detection of circulating thyroid specific transcripts (TSHR/Tg-mRNAs) in thyroid cancer patients: Their diagnostic significance

Thyroid cancer is the most prevalent endocrine malignancy. The preoperative diagnosis of differentiated thyroid cancer (DTC) that relies solely on fine-needle aspiration (FNAC) biopsy, sometimes possesses conflicting results. New molecular markers for thyroid cancer have been investigated with most of them based on the detection in thyroid nodules or tumor tissue specimens. Recently, it was possible to detect thyroid cancer cells in the circulation by measuring the mRNA of thyroid specific genes. Among these, thyroglobulin and more recently thyroid stimulating hormone receptor mRNAs, TSHR/Tg-mRNAs in peripheral blood might serve as cancer-specific markers. These have become promising new circulating markers for thyroid cancer. The purpose of this study is to assess TSHR/Tg-mRNAs as diagnostic molecular markers for thyroid cancer and if they can be used preoperatively in synergy with FNAC. This study was performed on 60 subjects; 20 healthy volunteers and 40 patients; including 16 patients with benign thyroid diseases, 24 patients with thyroid cancer; 18 patients with newly diagnosed (DTC) and 6 patients with recurrent thyroid cancer. Diagnosis of cancer was based on FNAC and histopathology of surgical specimens. All subjects had TSHR/Tg-mRNAs in peripheral blood measured by reverse transcriptase (RT)-PCR. Based on cytology/pathology; 18 patients had newly diagnosed DTC and 11 had benign thyroid disease. Preoperative FNAC was performed on 29 of 40 patients; FNAC was diagnostic in 11/18 of malignant lesions (61.1%), in 8/11 of benign lesions (72.7%), while 10/29 (34.5%) were indeterminate. TSHR/Tg-mRNAs correctly diagnosed DTC in 20/24 and 19/24 (sensitivity 83.3% and 79.1%) and benign disease in 14/16 and 13/16 (specificity 87.5% and 81.3%), respectively. With indeterminate FNA, TSHR/Tg-mRNAs correctly diagnosed DTC (follicular type) in 5/7 and benign disease in 2/3 (combined sensitivity 71.4%; specificity 66.7%). There was high concordance between RT-PCR results for TSHR-mRNA and Tg-mRNA. Of the controls 19/20 (95%) and 16/20 (80%) were negative for both TSHR- and Tg-mRNAs. With the use of a carefully selected primer pair and qualitative RT-PCR; our results indicate that TSHR/Tg-mRNAs in peripheral blood are both equally sensitive and specific markers for detection of thyroid cancer cells. Combining TSHR/Tg-mRNAs and FNAC and ultrasound enhances the preoperative detection of cancer in patients with thyroid nodules, reducing unnecessary surgeries and correctly classified most follicular cancers and could have spared surgery in patients with benign disease.

Peripheral thyrotropin receptor mRNA as a novel marker for differentiated thyroid cancer diagnosis and surveillance

Thyroid cancer is the most prevalent endocrine cancer whose incidence rates, particularly among women, have increased over the last decade. Although survival outcomes following surgery (with or without radioactive iodine ablation treatment) remain favorable, a significant proportion of patients are at lifetime risk of locoregional lymph node recurrence and distant metastasis. Serum thyroglobulin (Tg) has been the only circulating marker in routine use for detecting thyroid cancer recurrence, but it lacks sensitivity and is unreliable when Tg antibodies are present. New molecular markers for thyroid cancer have been investigated, with most based on detection in thyroid nodule or tumor tissue specimens. Recently, it has become possible to detect thyroid cancer cells in peripheral blood by measuring the mRNA of thyroid-specific genes, such as the mRNA of Tg and thyrotropin receptor. These have become promising new circulating markers for thyroid cancer. This review highlights the progress in this field from the perspective of improved initial cancer diagnosis and enhanced ability to monitor thyroid cancer recurrence.