Enhancing Early Relapse Detection in Testicular Cancer through Rolling Circle Amplification of microRNA Biomarkers

This project aims to improve the accuracy of early-stage testicular germ cell tumor (GCT) relapse detection in patients through rolling circle amplification (RCA) of plasma microRNA miR371. GCTs pose a significant challenge in clinical management due to the lack of sensitive and specific biomarkers for early relapse detection. Patients are at risk of over-treatment and long-term chemotherapy toxicity that negatively impacts life expectancy and quality of life. Patient blood samples from the BC provincial genitourinary biobank will be used.

 

Commonly used Polymerase Chain Reaction (PCR)-based detection methods have limitations such as precise temperature control requirements that other isothermal amplification techniques lack. RCA is an isothermal method that is effective in treatment and research applications. The clinical validity of microRNA miR371a-3p has been shown to be a potential biomarker for non-teratoma GCTs. However, the sensitivity of this biomarker in early-stage GCT detection using current PCR methods is low, and more accurate methods to detect smaller amounts of circulating miR371 are needed. RCA result sensitivity will be evaluated against PCR results.

 

The successful implementation of RCA will validate a more sensitive method for miR371 analysis. This will enhance clinical decision-making, reduce treatment toxicity, and extend the reach and applicability of early GCT relapse detection to resource-limited settings.
Andy Jia is a Master of Science student specializing in Interdisciplinary Oncology at the University of British Columbia (UBC). The project will be conducted in the Nappi Lab at the Vancouver Prostate Centre, which has extensive experience in miR371 analysis.

 

The Canadian Consortium of Clinical Trial Training Platform (CANTRAIN) and Michael Smith Health Research BC provide funding through the CANTRAIN-CTTP & Michael Smith Health Research BC Master’s Studentship 2023 Award Program.

End of Award Update – December 2024

 

Results

Our promising preliminary results demonstrated that the RCA method can successfully amplify and detect miR371 samples. Our findings indicate that synthetic miR371 was amplified to significant concentrations, and optimizations to our probe designs improved our results with each modification.

 

Impact

The project relies on plasma samples from patients enrolled in the provincial Genitourinary Cancers (GU) biobank that have already been collected and analyzed. Hence, the findings of this project can profoundly impact the clinical management of testicular cancer. RCA can be easily implemented in resource-limited settings, including rural areas and developing countries, extending its reach and applicability beyond the laboratory setting.

 

Potential Influence

The impact of this research project is expected to be significant. The primary expected result is validating a new sensitive method for miR371 analysis that could predict tumor relapse in patients with clinical stage 1 testicular germ cell tumors. This validation will enhance clinical decision-making, improve patient management, and reduce unnecessary treatments, leading to reduced healthcare expenses in British Columbia.

 

Next Steps

Continued optimization of the RCA protocol will be implemented. Patient samples will be used extensively, and to reach a lower temperature of RCA incubation and ensure accurate detection, new probes will be designed, and different reagents will be tested. RCA implementation with electrochemical biosensors for point-of-care detection will be investigated in depth when a lower temperature of RCA amplification has been successfully reached.

 

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