Current evidence shows that serum miR-371a-3p can identify disease recurrence in testicular germ cell tumour (TGCT) patients and correlates with tumour load. Despite convincing evidence showing the advantages of including miR-371a-3p testing to complement and overcome the classical serum tumour markers limitations, the successful introduction of a serum miRNA based test into clinical practice has been impeded by a lack of consensus regarding optimal methodologies and lack of a universal protocol and thresholds. Herein, we investigate two quantitative real-time PCR (qRT-PCR) based pipelines in detecting disease recurrence in stage I TGCT patients under active surveillance, and compare the sensitivity and specificity for each method.
Sequential serum samples collected from 33 stage I TGCT patients undergoing active surveillance were analysed for miR-371a-3p via qRT-PCR with and without an amplification step included.
Using a pre-amplified protocol, all known recurrences were detected via elevated miR-371a-3p expression, while without pre-amplification, we failed to detect recurrence in 3/10 known recurrence patients. For pre-amplified analysis, sensitivity and specificity was 90% and 94.4% respectively. Without amplification, sensitivity dropped to 60%, but exhibited 100% specificity.
We conclude that incorporating pre-amplification increases sensitivity of miR-371a-3p detection, but produces more false positive results. The ideal protocol for quantification of miR-371a-3p still needs to be determined. TGCT patients undergoing active surveillance may benefit from serum miR-371a-3p quantification with earlier detection of recurrences compared to current standard methods. However, larger cross-institutional studies where samples are processed and data is analysed in a standardised manner are required prior to its routine clinical implementation.
Frontiers in oncology. 2022 Dec 08*** epublish ***
Ailsa J Christiansen, João Lobo, Christian D Fankhauser, Christian Rothermundt, Richard Cathomas, Aashil A Batavia, Josias B Grogg, Arnoud J Templeton, Anita Hirschi-Blickenstorfer, Anja Lorch, Silke Gillessen, Holger Moch, Jörg Beyer, Thomas Hermanns
Department of Urology, University Hospital Zurich, University of Zurich, Zurich, Switzerland., Cancer Biology and Epigenetics Group, Research Center of Portuguese Institute of Oncology (IPO) Porto, RISE@CI-IPOP Health Research Network, Portuguese Oncology Institute of Porto, Porto Comprehensive Cancer Center, Porto, Portugal., University of Zurich, Zurich, Switzerland., Department of Oncology, Kantonsspital, St. Gallen, Switzerland., Division of Oncology/Hematology, Kantonsspital Graubünden, Chur, Switzerland., Department of Pathology and Molecular Pathology, University Hospital Zurich, University of Zurich, Zurich, Switzerland., St. Clara Research, St. Claraspital Basel and Faculty of Medicine, University of Basel, Basel, Switzerland., Onkozentrum Hirslanden, Klinik Hirslanden, Zürich, Switzerland., Department of Oncology, University Hospital Zurich, University of Zurich, Zurich, Switzerland., Department of Medical Oncology, Inselspital, University Hospital of Bern, Bern, Switzerland.