Multicenter Evaluation of Independent High-Throughput and RT-qPCR Technologies for the Development of Analytical Workflows for Circulating miRNA Analysis
Autor: | Martin H D Neumann, Vera Kloten, Rita Lampignano, Thomas Schlange, Klaus Pantel, Martin Schlumpberger, Melanie Janning, Francesca Di Pasquale, Mikael Kubista, Sonja Loges, Anna Babayan, Franca Kobus, Andrei Herdean, Jonathan M Shaffer, Thomas Krahn, Markus Sprenger-Haussels |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
Oncology Cancer Research medicine.medical_specialty multicentric study Context (language use) lcsh:RC254-282 Article 03 medical and health sciences 0302 clinical medicine Internal medicine microRNA medicine Blood test 1112 Oncology and Carcinogenesis Liquid biopsy Lung cancer high-throughput medicine.diagnostic_test liquid biopsy business.industry Confounding RT-qPCR medicine.disease lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens Biomarker (cell) circulating miRNA 030104 developmental biology Real-time polymerase chain reaction 030220 oncology & carcinogenesis business |
Zdroj: | Cancers, Vol 12, Iss 1166, p 1166 (2020) Cancers Volume 12 Issue 5 |
ISSN: | 2072-6694 |
Popis: | Background: Among emerging circulating biomarkers, miRNA has the potential to detect lung cancer and follow the course of the disease. However, miRNA analysis deserves further standardization before implementation into clinical trials or practice. Here, we performed international ring experiments to explore (pre)-analytical factors relevant to the outcome of miRNA blood tests in the context of the EU network CANCER-ID. Methods: Cell-free (cfmiRNA) and extracellular vesicle-derived miRNA (EVmiRNA) were extracted using the miRNeasy Serum/Plasma Advanced, and the ExoRNeasy Maxi kit, respectively, in a plasma cohort of 27 NSCLC patients and 20 healthy individuals. Extracted miRNA was investigated using small RNA sequencing and hybridization platforms. Validation of the identified miRNA candidates was performed using quantitative PCR. Results: We demonstrate the highest read counts in healthy individuals and NSCLC patients using QIAseq. Moreover, QIAseq showed 15.9% and 162.9% more cfmiRNA and EVmiRNA miRNA counts, respectively, in NSCLC patients compared to healthy control samples. However, a systematic comparison of selected miRNAs revealed little agreement between high-throughput platforms, thus some miRNAs are detected with one technology, but not with the other. Adding to this, 35% (9 of 26) of selected miRNAs in the cfmiRNA and 42% (11 of 26) in the EVmiRNA fraction were differentially expressed by at least one qPCR platform about half of the miRNAs (54%) were concordant for both platforms. Conclusions: Changing of (pre)-analytical methods of miRNA analysis has a significant impact on blood test results and is therefore a major confounding factor. In addition, to confirm miRNA biomarker candidates screening studies should be followed by targeted validation using an independent platform or technology. |
Databáze: | OpenAIRE |
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