Abstract A005: Exploring liquid biopsy for the detection of bladder cancer: Unraveling RNA fusions
Authors: Mayer Saidian, Jason Saenz, Carlos Hernandez, Lauren Lee, Andrew Dunnigan, Yabin Lu, Nafiseh Jafari
Journal: Clinical Cancer Research (2024)

Abstract Introduction: This study delves into the promising realm of liquid biopsy for the non-invasive detection of bladder cancer. Leveraging urine as a valuable source, our investigation focuses on identifying RNA fusions associated with bladder cancer, with specific attention to ETV6-NTRK3 and PRCC-TFE3. The utilization of liquid biopsy offers a less invasive and more accessible approach to diagnosing bladder cancer, potentially revolutionizing the field of cancer diagnostics. These specific RNA fusions in urine samples may serve as reliable biomarkers for early detection and monitoring of bladder cancer. This research contributes to the ongoing efforts to enhance diagnostic methodologies to improve patient outcomes and streamline cancer management. Materials and Methods: Urine was collected from healthy male donors using UAS, a urine preservative from Novosanis, for improved urine preservation and stability. The urine samples with UAS were centrifuged at 16,000 xg for 10 min to obtain cf-urine. Three 20 mL aliquots of the precleared supernatant urine samples were spiked with an RNA fusion reference standard from Anchor Molecular containing ETV6-NTRK3 and PRCC-TFE3RNA at a concentration of 20ng/mL. All samples were extracted using the nRichDX Revolution Max20 cfTNA Isolation Kit. The eluants from the extracted samples were DNAse treated. The extracted cfTNAs profile was assessed on 4150 TapeStation System, Agilent, using RNA ScreenTape. Extracted nucleic acids from urine were evaluated using a RT-qPCR assay to quantify the number of copies of the fusion RNAs. Results: The Renal RNA fusions ETV6-NTRK3 and PRCC-TFE3RNA were successfully extracted using the nRichDX Revolution cfTNA Isolation Kit. TapeStation analysis demonstrated prominent 18S and 28S peaks, confirming the successful extraction of RNA from 20 mL urine samples. The mean percent recovery of the recovered RNA Fusion mutation was about 70% via RT-qPCR. Conclusion: Our findings indicate a recovery rate of approximately 70% for spiked RNA fusions in the urine samples. This research holds the promise of revolutionizing cancer diagnostics by introducing a method that is not only reliable but also convenient for early detection and monitoring. Identifying specific RNA fusions in urine samples as biomarkers is a significant step in enhancing diagnostic methodologies. As we contribute to these ongoing efforts, we aim to improve patient outcomes and streamline the management of bladder cancer through innovative and effective diagnostic approaches. Citation Format: Mayer Saidian, Jason Saenz, Carlos Hernandez, Lauren Lee, Andrew Dunnigan, Yabin Lu, Nafiseh Jafari. Exploring liquid biopsy for the detection of bladder cancer: Unraveling RNA fusions [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr A005.