Parallel Liquid Biopsy Analysis of miR‐371a‐3p and Cell‐Free DNA in an Unselected Cohort of Patients With Testicular Germ Cell Tumors
Laura Matouskova, Romy Kralova, Eva Parobkova, Violeta Bakardjieva‐Mihaylova, Martina Zwyrtkova, Karolina Skvarova Kramarzova, Ludmila BoublikovaABSTRACT
Background
Despite robust data showing strong potential to improve diagnosis and disease monitoring in testicular germ cell tumors (TGCTs), miR‐371a‐3p testing has not yet been implemented in routine clinical practice. Cell‐free DNA (cfDNA) represents another promising liquid biopsy biomarker that may provide complementary molecular information.
Objectives
To evaluate miR‐371a‐3p expression in an unselected TGCT patient cohort, compare its performance with simultaneous analysis of cfDNA levels, and with clinical and laboratory data to address their impact on the TGCT management.
Materials and Methods
In total, 119 prospectively collected plasma samples from 77 unselected TGCT patients treated at a single center were analyzed. miR‐371a‐3p was detected using a commercial assay, normalized to miR‐30b‐5p as an internal control. cfDNA levels were quantified by qPCR using albumin as a housekeeping gene.
Results
miR‐371a‐3p was present in 19 of 112 (17%) evaluable samples, with no significant correlation to cfDNA levels ( p > 0.05). The sensitivity and specificity for active disease detection were 38% and 95%, respectively, with receiver operating characteristic and area under the curve 0.6619, p = 0.0217. miR‐371a‐3p was detected at a much higher frequency in samples from the time points of active malignant disease ( p = 0.0014) or with positive serum tumor markers ( p = 0.012) than in samples from remission. On the contrary, total cfDNA levels were highest in samples taken during the chemotherapy course ( p < 0.0001).
Conclusion
miR‐371a‐3p and cfDNA provide independent information in TGCT, miR‐371a‐3p being associated with the active disease presence, while total cfDNA levels are increased during the active treatment, reflecting different biology and dynamics of the two circulating biomarkers. These findings have implications for future studies on disease monitoring in TGCT. The sensitivity of miR‐371a‐3p detection in this real practice cohort is lower than reported previously, suggesting assay standardization and clinical studies setting are warranted before miR‐371a‐3p implementation into clinical practice.