Validity of multiplex biomarker model of 6 genes for the differential diagnosis of thyroid nodules

Abstract

BackgroundCurrently the cytological examination of fine needle aspiration (FNA) biopsies is the standard technique for the pre-operative differential diagnosis of thyroid nodules. However, the results may be non-informative in ~20% of cases due to an inadequate sampling and the lack of highly specific, measurable cytological criteria, therefore ancillary biomarkers that could aid in these cases are clearly needed. The aim of our study was to evaluate the mRNA expression levels of 8 candidate marker genes as the diagnostic biomarkers for the discrimination of benign and malignant thyroid nodules and to find a combination of biomarkers with the highest diagnostic value.Materials and methodsmRNA expression levels of eight candidate marker genes - BIRC5, CCND1, CDH1, CITED1, DPP4, LGALS3, MET and TFF3 was measured by real-time RT-PCR in paired nodular and surrounding normal thyroid tissue specimens of 105 consecutive patients undergoing thyroid surgery and compared between different types of thyroid lesions.ResultsSignificant differences in the mRNA expression levels between the normal and malignant thyroid tissues and between benign and malignant nodules were found for BIRC5, CCND1, CITED1, DPP4, LGALS3, MET and TFF3, but not CDH1. On a single gene basis, relative quantity (RQ) of LGALS3 had the highest diagnostic value for the discrimination of malignant and benign thyroid nodules (AUC = 0.832, P < 0.0001 and 90.9% sensitivity and 65.6% specificity at the optimal cut-off on ROC curve). The only two-marker set that outperformed LGALS3 was RQ sum of LGALS3 and BIRC5 (AUC = 0.841, P < 0.0001). An application of multivariate logistic regression analysis resulted in the generation of a multiplex biomarker model based on LGALS3, BIRC5, TFF3, CCND1, MET and CITED1 that had considerably higher specificity than a single marker or two marker gene-based models (AUC = 0.895, P < 0.0001, 70.5% sensitivity and 93.4% specificity).ConclusionsThis study confirmed that mRNA expression levels of 7 out of 8 candidate genes analysed have a diagnostic value for the distinction of benign and malignant thyroid nodules. The multiplex biomarker model based on 6 genes outperformed a single marker or two marker-based models and warrants feasibility studies on FNA biopsies and the validation in a larger cohort of patients.

A three‐gene panel that distinguishes benign from malignant thyroid nodules

“…DPP4 (CD26) encodes a membrane‐bound serine protease, a member of the dipeptidyl prolyl peptidase (DPP) family . Our findings are supported by literature reports that this gene significant increases in differentiated carcinomas vs. normal or benign thyroid nodules at average 46.19 times . In addition, some studies showed that DPP4 was downregulated in ATC with unclear mechanism, which was consistent with the expression level of our ACT specimen by QPCR.…”

Integration of Traditional and Metabolomics Biomarkers Identifies Prognostic Metabolites for Predicting Responsiveness to Nutritional Intervention against Oxidative Stress and Inflammation

“…Several other studies have supported an association between urinary glycine or PAG levels and status of oxidative stress [47,48,49,50,51,52] or inflammatory stress [52]. Finally, the predictive accuracy of this two-metabolite set was further validated with the LOOCV procedure, because the ROC curve may lead to an overestimation of the AUC if it is constructed using all samples [53]. The LOOCV procedure has an advantage of reducing the likelihood of developing an overly optimistic predictive model given the relatively small sample in our study [54].…”

Measurement of <i>TFF3</i> mRNA in aspirates from thyroid nodules using mesh filtration: The first clinical trial in 130 cases

“…In the follow-up studies, many researchers at other institutes confirmed the utility of TFF3 mRNA in diagnosing follicular carcinoma [10][11][12][13][14][15][16][17]. A more intriguing property of TFF3 mRNA is that papillary and anaplastic thyroid carcinomas also show low T/G ratios [9,18].…”

“…However, in the case of TFF3 mRNA, the precise measurement of its expression levels in thyroid tumor cells in aspirates was difficult because it is interfered with peripheral blood cells or infiltrating lymphocytes that are aspirated simultaneously by FNAB. In fact, there has been only a few reports measuring TFF3 mRNA using thyroid aspirates [13][14][15][16]. Thus, selection of thyroid tumor cells in aspirated samples is crucial for the precise measurement of the T/G ratio in aspirates.…”