Spatial Distribution Patterns of Clinically Relevant TERT Promoter Mutations in Follicular Thyroid Tumors of Uncertain Malignant Potential

Hysek, Martin; Jatta, Kenbugul; Hellgren, Laila; Stenman, Adam; Zedenius, Jan; Juhlin, Carl Christofer

doi:10.1016/j.jmoldx.2020.10.016

Cited by 13 publications

(13 citation statements)

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“…Evaluation of final surgical specimen with ddPCR would provide further insight into the utility of FNAB. This is especially true in genetically heterogenous tumors, where location of FNAB directly affects results [ 43 , 44 ]. A limitation of evaluating TERT expression by ddPCR is that lymphocytes are also known to express TERT, and therefore lymphocyte contamination of the tumor sample could theoretically produce a false positive result [ 45 ].…”

Section: Discussionmentioning

confidence: 99%

Utility of droplet digital polymerase chain reaction for TERT and BRAF mutational profiling of thyroid nodules

et al. 2021

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Background Mutations involving BRAF and TERT are important predictors of disease severity in thyroid cancer, but molecular testing is limited by cost and lack of adequate tissue sample. This study aimed to assess the utility of BRAFV600E and TERT testing using droplet digital PCR (ddPCR) as a diagnostic and prognostic tool for thyroid fine needle aspirate biopsy (FNAB). Methods Patients with thyroid nodules were prospectively enrolled from March 2015 to September 2018. Pre-operative FNAB was collected for standard cytology and molecular testing. BRAFV600E and TERT levels were analyzed by ddPCR. Cytology (Bethesda system) and ddPCR results were correlated to surgical pathology. Results A total of 222 patients were enrolled, of which 124 received thyroid surgery. Pre-operative cytology alone with Bethesda ≥5 was 100% specific and 70% sensitive for malignancy on final surgical pathology. BRAFV600E positivity or TERT overexpression was 100% specific and 60.0% sensitive. Combining cytology (Bethesda ≥5) with BRAFV600E and TERT testing increased the sensitivity of a malignant diagnosis to 80.0%. High TERT levels and/or BRAFV600E was associated with aggressive or advanced stage pathology. Conclusions Combining cytology with ddPCR analysis of BRAFV600E and TERT can improve the diagnostic accuracy of thyroid FNAB, and help predict aggressive pathology.

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Section: Discussionmentioning

confidence: 99%

Utility of droplet digital polymerase chain reaction for TERT and BRAF mutational profiling of thyroid nodules

et al. 2021

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“…This would explain why hTERT mRNA levels affect prognosis in almost all tumors but a singly evaluated mechanism could do it or not [9,[13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32]. Additionally, TERT promoter mutations have been shown to be spatially heterogeneous/sub-clonal in some tumors (follicular thyroid carcinoma, follicular thyroid tumors of uncertain malignant potential and meningiomas) and several genes (MLH1, MGMT, CDKN2B) could exhibit spatially heterogeneous/sub-clonal methylation patterns in different tumors (glioblastoma, breast carcinoma) [52][53][54][55][56][57][58]. Although these aspects have never been investigated in MCC, it is not possible to exclude that mhTERT and promoter methylation could be spatially heterogeneous/sub-clonal in this tumor and, as result, our data could be affected by the tissue/block selection.…”

Section: Discussionmentioning

confidence: 99%

Intron 4–5 hTERT DNA Hypermethylation in Merkel Cell Carcinoma: Frequency, Association with Other Clinico-pathological Features and Prognostic Relevance

Ricci

Morandi

Ambrosi³

et al. 2021

Endocr Pathol

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Merkel cell carcinoma (MCC) is an aggressive skin tumor with neuroendocrine differentiation, mainly affecting elderly population or immunocompromised individuals. As methylation of the human telomerase reverse transcriptase (mhTERT) has been shown to be a prognostic factor in different tumors, we investigated its role in MCC, in particular in intron 4–5 where rs10069690 has been mapped and recognized as a cancer susceptibility locus. DNA methylation analysis of hTERT gene was assessed retrospectively in a cohort of 69 MCC patients from the University of Bologna, University of Turin and University of Insubria. Overall mortality was evaluated with Kaplan-Meier curves and multivariable Royston-Parmar models. High levels of mhTERT (mhTERThigh) (HR = 2.500, p = 0.015) and p63 (HR = 2.659, p = 0.016) were the only two clinico-pathological features significantly associated with a higher overall mortality at the multivariate analysis. We did not find different levels of mhTERT between MCPyV (+) and (−) cases (21 vs 14, p = 0.554); furthermore, mhTERThigh was strongly associated with older age (80.5 vs 72 years, p = 0.026), no angioinvasion (40.7% vs 71.0%, p = 0.015), lower Ki67 (50 vs 70%, p = 0.005), and PD-L1 expressions in both tumor (0 vs 3%, p = 0.021) and immune cells (0 vs 10%, p = 0.002). mhTERT is a frequently involved epigenetic mechanism and a relevant prognostic factor in MCC. In addition, it belongs to the shared oncogenic pathways of MCC (MCPyV and UV-radiations) and it could be crucial, together with other epigenetic and genetic mechanisms as gene amplification, in determining the final levels of hTERT mRNA and telomerase activity in these patients.

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“…Mutation analysis of FFPE samples was performed in the same laboratory under clinical routine diagnostic circumstances, using a previously published methodology for DNA extraction and ddPCR analysis of TERT. 26…”

Section: Dna Extraction and Digital Droplet Pcr (Ddpcr) Analyses Of T...mentioning

confidence: 99%

“…25 However, previous data suggest that digital droplet PCR (ddPCR) may be an even more sensitive method to detect sub-clonal TERT promoter mutations compared to conventional Sanger sequencing in excised thyroid tumors. 26 In ddPCR, the DNA material is separated into thousands of nanoliter-sized water-in-oil droplets and then interrogated for the variant of interest. By adding fluorescence-labeled probes targeting either the wild-type or the mutated allele, the results can be visualized as points on a graph.…”

Section: Introductionmentioning

confidence: 99%

Digital droplet PCRTERT promoter mutational screening in fine needle aspiration cytology of thyroid lesions: A highly specific technique for pre‐operative identification of high‐risk cases

Hysek

Hellgren

Stenman

et al. 2023

Diagnostic Cytopathology

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Background Despite the advent of comprehensive molecular testing in surgical pathology, most centers still rely on the morphological assessment of fine‐needle aspiration cytology (FNAC) to triage patients with thyroid nodules for surgery. Subsets of patients could benefit from the inclusion of molecular testing to increase the diagnostic and/or prognostic properties of the cytology analysis, including the assessment of TERT promoter mutations, an event coupled with thyroid malignancy, and poor prognosis. Methods In this prospective study, preoperative FNAC material from 65 cases was assessed for TERT promoter hotspot mutations C228T and C250T using the digital droplet PCR (ddPCR) technique on frozen pellets and re‐evaluated postoperatively. Results Our cohort consisted of 15 B‐III (23%), 26 B‐IV (40%), 1 B‐V (2%), and 23 (35%) B‐VI lesions according to the Bethesda System for Reporting Thyroid Cytopathology. TERT promoter mutations were detected in 7 cases; 4 papillary thyroid carcinomas (all with preoperative B‐VI status), two follicular thyroid carcinomas (one B‐IV and one B‐V status), and one poorly differentiated thyroid carcinoma (with B‐VI status). All mutated cases were verified by mutational analysis of tumor tissue derived from postoperative formalin‐fixed paraffin‐embedded tissue, while all cases identified as wild‐type on FNAC remained wild‐type postoperatively. Moreover, the occurrence of a TERT promoter mutation was significantly associated with malignant disease and higher Ki‐67 proliferation indices. Conclusion In the present cohort, we found that ddPCR is a highly specific method for detecting high‐risk TERT promoter mutations on thyroid FNAC material that could guide different surgical approaches in subsets of indeterminate lesions if reproduced in larger materials.

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Spatial Distribution Patterns of Clinically Relevant TERT Promoter Mutations in Follicular Thyroid Tumors of Uncertain Malignant Potential

Cited by 13 publications

References 40 publications

Utility of droplet digital polymerase chain reaction for TERT and BRAF mutational profiling of thyroid nodules

Utility of droplet digital polymerase chain reaction for TERT and BRAF mutational profiling of thyroid nodules

Intron 4–5 hTERT DNA Hypermethylation in Merkel Cell Carcinoma: Frequency, Association with Other Clinico-pathological Features and Prognostic Relevance

Digital droplet PCRTERT promoter mutational screening in fine needle aspiration cytology of thyroid lesions: A highly specific technique for pre‐operative identification of high‐risk cases

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