Evaluation of TERT promoter mutations in urinary cell-free DNA and sediment DNA for detection of bladder cancer

2019 ◽  
Vol 64 ◽  
pp. 60-63 ◽  
Author(s):  
Sebastian Stasik ◽  
Karsten Salomo ◽  
Ulrike Heberling ◽  
Michael Froehner ◽  
Ulrich Sommer ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cell Free Dna ◽  
Free Dna ◽  
Tert Promoter ◽  
Tert Promoter Mutations ◽  
Promoter Mutations

scholarly journals Development of Sensitive Droplet Digital PCR Assays for Detecting Urinary TERT Promoter Mutations as Non-Invasive Biomarkers for Detection of Urothelial Cancer

Cancers ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 3541 ◽  
Author(s):  
Md Ismail Hosen ◽  
Nathalie Forey ◽  
Geoffroy Durand ◽  
Catherine Voegele ◽  
Selin Bilici ◽  
...  
Keyword(s):  
Urothelial Cancer ◽  
Digital Pcr ◽  
Kappa Coefficient ◽  
Droplet Digital Pcr ◽  
Cell Free Dna ◽  
Non Invasive ◽  
Free Dna ◽  
Tert Promoter ◽  
Tert Promoter Mutations ◽  
Promoter Mutations

Somatic mutations in the telomerase reverse transcriptase (TERT) promoter regions are frequent events in urothelial cancer (UC) and their detection in urine (supernatant cell-free DNA or DNA from exfoliated cells) could serve as putative non-invasive biomarkers for UC detection and monitoring. However, detecting these tumor-borne mutations in urine requires highly sensitive methods, capable of measuring low-level mutations. In this study, we developed sensitive droplet digital PCR (ddPCR) assays for detecting TERT promoter mutations (C228T, C228A, CC242-243TT, and C250T). We tested the C228T and C250T ddPCR assays on all samples with sufficient quantity of urinary DNA (urine supernatant cell-free DNA (US cfDNA) or urine pellet cellular DNA (UP cellDNA)) from the DIAGURO (n = 89/93 cases and n = 92/94 controls) and from the IPO-PORTO (n = 49/50 cases and n = 50/50 controls) series that were previously screened with the UroMuTERT assay and compared the performance of the two approaches. In the DIAGURO series, the sensitivity and specificity of the ddPCR assays for detecting UC using either US cfDNA or UP cellDNA were 86.8% and 92.4%. The sensitivity was slightly higher than that of the UroMuTERT assay in the IPO-PORTO series (67.4% vs. 65.3%, respectively), but not in the DIAGURO series (86.8% vs. 90.7%). The specificity was 100% in the IPO-PORTO controls for both the UroMuTERT and ddPCR assays, whereas in the DIAGURO series, the specificity dropped for ddPCR (92.4% versus 95.6%). Overall, an almost perfect agreement between the two methods was observed for both US cfDNA (n = 164; kappa coefficient of 0.91) and UP cellDNA (n = 280; kappa coefficient of 0.94). In a large independent series of serial urine samples from DIAGURO follow-up BC cases (n = 394), the agreement between ddPCR and UroMuTERT was (i) strong (kappa coefficient of 0.87), regardless of urine DNA types (kappa coefficient 0.89 for US cfDNA and 0.85 for UP cellDNA), (ii) the highest for samples with mutant allelic fractions (MAFs) > 2% (kappa coefficient of 0.99) and (iii) only minimal for the samples with the lowest MAFs (< 0.5%; kappa coefficient 0.32). Altogether, our results indicate that the two methods (ddPCR and UroMuTERT) for detecting urinary TERT promoter mutations are comparable and that the discrepancies relate to the detection of low-allelic fraction mutations. The simplicity of the ddPCR assays makes them suitable for implementation in clinical settings.

scholarly journals Sensitive droplet digital PCR method for detection of TERT promoter mutations in cell free DNA from patients with metastatic melanoma

Oncotarget ◽  
2017 ◽  
Vol 8 (45) ◽  
pp. 78890-78900 ◽  
Author(s):  
Ashleigh C. McEvoy ◽  
Leslie Calapre ◽  
Michelle R. Pereira ◽  
Tindaro Giardina ◽  
Cleo Robinson ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Cell Free Dna ◽  
Free Dna ◽  
Tert Promoter ◽  
Tert Promoter Mutations ◽  
Pcr Method ◽  
Promoter Mutations

scholarly journals The role of telomerase reverse transcriptase (TERT) promoter mutations in prognosis in bladder cancer

Bioengineered ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 1495-1504
Author(s):  
Song Wan ◽  
Xuan Liu ◽  
Wei Hua ◽  
Ming Xi ◽  
Yulin Zhou ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Reverse Transcriptase ◽  
Telomerase Reverse Transcriptase ◽  
Tert Promoter ◽  
Tert Promoter Mutations ◽  
Promoter Mutations

scholarly journals Urinary TERT promoter mutations are detectable up to 10 years prior to clinical diagnosis of bladder cancer: Evidence from the Golestan Cohort Study

EBioMedicine ◽  
2020 ◽  
Vol 53 ◽  
pp. 102643 ◽  
Author(s):  
Md Ismail Hosen ◽  
Mahdi Sheikh ◽  
Maria Zvereva ◽  
Ghislaine Scelo ◽  
Nathalie Forey ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cohort Study ◽  
Clinical Diagnosis ◽  
Tert Promoter ◽  
Tert Promoter Mutations ◽  
Promoter Mutations

scholarly journals Urine TERT promoter mutations‑based tumor DNA detection in patients with bladder cancer: A pilot study

2021 ◽  
Vol 15 (6) ◽  
Author(s):  
Mark Jain ◽  
David Kamalov ◽  
Alexander Tivtikyan ◽  
Alexander Balatsky ◽  
Larisa Samokhodskaya ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Pilot Study ◽  
Dna Detection ◽  
Tert Promoter ◽  
Tert Promoter Mutations ◽  
Promoter Mutations ◽  
Tumor Dna

scholarly journals TERT Promoter Mutation Analysis of Whole-Organ Mapping Bladder Cancers

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 230
Author(s):  
Veronika Weyerer ◽  
Markus Eckstein ◽  
Pamela L. Strissel ◽  
Adrian Wullweber ◽  
Fabienne Lange ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Telomere Length ◽  
Hot Spot ◽  
Tumor Development ◽  
Non Invasive ◽  
Tert Promoter ◽  
Mutational Status ◽  
Tert Promoter Mutations ◽  
Promoter Mutations ◽  
Normal Urothelium

Background: Multifocal occurrence is a main characteristic of urothelial bladder cancer (UBC). Whether urothelial transformation is caused by monoclonal events within the urothelium, or by polyclonal unrelated events resulting in several tumor clones is still under debate. TERT promoter mutations are the most common somatic alteration identified in UBC. In this study, we analyzed different histological tissues from whole-organ mapping bladder cancer specimens to reveal TERT mutational status, as well as to discern how tumors develop. Methods: Up to 23 tissues from nine whole-organ mapping bladder tumor specimens, were tested for TERT promoter mutations including tumor associated normal urothelium, non-invasive urothelial lesions (hyperplasia, dysplasia, metaplasia), carcinoma in situ (CIS) and different areas of muscle invasive bladder cancers (MIBC). The mutational DNA hotspot region within the TERT promoter was analyzed by SNaPshot analysis including three hot spot regions (−57, −124 or −146). Telomere length was measured by the Relative Human Telomere Length Quantification qPCR Assay Kit. Results: TERT promoter mutations were identified in tumor associated normal urothelium as well as non-invasive urothelial lesions, CIS and MIBC. Analysis of separate regions of the MIBC showed 100% concordance of TERT promoter mutations within a respective whole-organ bladder specimen. Polyclonal events were observed in five out of nine whole-organ mapping bladder cancers housing tumor associated normal urothelium, non-invasive urothelial lesions and CIS where different TERT promoter mutations were found compared to MIBC. The remaining four whole-organ mapping bladders were monoclonal for TERT mutations. No significant differences of telomere length were observed. Conclusions: Examining multiple whole-organ mapping bladders we conclude that TERT promoter mutations may be an early step in bladder cancer carcinogenesis as supported by TERT mutations detected in tumor associated normal urothelium as well as non-invasive urothelial lesions. Since mutated TERT promoter regions within non-invasive urothelial lesions are not sufficient alone for the establishment of cancerous growth, this points to the contribution of other gene mutations as a requirement for tumor development.

scholarly journals Liquid Biopsy Analysis of FGFR3, TERT Promoter and STAG2 Hotspot Mutations for Disease Surveillance in Bladder Cancer

2020 ◽  
pp. 1-7
Author(s):  
Victor Romanov ◽  
Dimitri Gnatenko ◽  
Edward Forsyth ◽  
Liang Xiaohui ◽  
Olga Povcher ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Sensitivity And Specificity ◽  
Liquid Biopsy ◽  
Disease Surveillance ◽  
Digital Pcr ◽  
Cell Free Dna ◽  
Free Dna ◽  
Tert Promoter ◽  
Mutational Status ◽  
Muscle Invasive

Patients with non-muscle invasive bladder cancer (NMIBC) are followed by frequent cystoscopies. Innovative approaches partly replacing cystoscopy (uncomfortable, expensive, low sensitive procedure) are demanded. The current study aims to establish a fast, reliable, non-invasive, and inexpensive procedure for NMIBC patient surveillance. Liquid biopsy is a reliable source of biomarkers for cancer patient monitoring. Urine is the most suitable biological liquid to search for bladder cancer biomarkers. Cell-free DNA in urine represents tumor-related mutations for several cancers, including the bladder. We investigated mutations in FGFR3, TERT promoter, and STAG2 as markers for diagnostics and follow-up in NMIBC. Digital PCR was used to detect mutations in urine-derived cell-free DNA. The sensitivity and specificity of the markers in relation to clinical outcomes served as criteria of the assay efficiency. The sensitivity with a single marker (TERT) reached 87%, with a specificity of 77%. Combining two biomarkers (TERT+FGFR3) increased the specificity of the assay to 100% with a sensitivity of 72%. Different mutational status of STAG2 can indicate NMIBC presence or recurrence. Therefore, applying the suggested combination of biomarkers with simple detection procedures to larger patient cohorts will allow developing procedures for BC detection and surveillance with optimal sensitivity and specificity. Based on the results of this proof-in-concept study, we conclude that this simple, fast and inexpensive assay can add diagnostic and prognostic value to cystoscopy/cytology analysis of NMIBC patients.

scholarly journals GABPA is a master regulator of luminal identity and restrains aggressive diseases in bladder cancer

2019 ◽  
Vol 27 (6) ◽  
pp. 1862-1877 ◽  
Author(s):  
Yanxia Guo ◽  
Xiaotian Yuan ◽  
Kailin Li ◽  
Mingkai Dai ◽  
Lu Zhang ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Transcription Factors ◽  
Gene Methylation ◽  
Tert Promoter ◽  
Tumorigenic Potential ◽  
Tert Promoter Mutations ◽  
Geo Dataset ◽  
Ets Family ◽  
Promoter Mutations

AbstractTERT promoter mutations occur in the majority of glioblastoma, bladder cancer (BC), and other malignancies while the ETS family transcription factors GABPA and its partner GABPB1 activate the mutant TERT promoter and telomerase in these tumors. GABPA depletion or the disruption of the GABPA/GABPB1 complex by knocking down GABPB1 was shown to inhibit telomerase, thereby eliminating the tumorigenic potential of glioblastoma cells. GABPA/B1 is thus suggested as a cancer therapeutic target. However, it is unclear about its role in BC. Here we unexpectedly observed that GABPA ablation inhibited TERT expression, but robustly increased proliferation, stem, and invasive phenotypes and cisplatin resistance in BC cells, while its overexpression exhibited opposite effects, and inhibited in vivo metastasizing in a xenograft transplant model. Mechanistically, GABPA directly activates the transcription of FoxA1 and GATA3, key transcription factors driving luminal differentiation of urothelial cells. Consistently, TCGA/GEO dataset analyses show that GABPA expression is correlated positively with luminal while negatively with basal signatures. Luminal tumors express higher GABPA than do basal ones. Lower GABPA expression is associated with the GABPA gene methylation or deletion (especially in basal subtype of BC tumors), and predicted significantly shorter patient survival based on TCGA and our cohort of BC patient analyses. Taken together, GABPA dictates luminal identity of BC cells and inhibits aggressive diseases in BC by promoting cellular differentiation despite its stimulatory effect on telomerase/TERT activation. Given these biological functions and its frequent methylation and/or deletion, GABPA serves as a tumor suppressor rather than oncogenic factor in BC. The GABPA effect on oncogenesis is context-dependent and its targeting for telomerase inhibition in BC may promote disease metastasizing.

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