Grant: Project Grant

The main concept of liquid biopsies for cancer monitoring is the detection of cancer-derived extracellular vesicles (EVs), which often have the same set of biomarkers as cancer cells. However, cancer-derived EVs typically have low concentration in the localized disease setting, which makes detection and quantification challenging. Ultracentrifugation (UC) is the most utilized EV-enriching technique, but it is known to co-isolate impurities, expose EVs to high shear stresses, and lead to poor quality of isolated EVs for analyses. We have created a technology that utilizes an Aqueous Two-Phase System (ATPS) fluid to isolate EVs and their genetic cargo, ctDNA and RNA, from biofluids, while maintaining the integrity of the isolated EVs. This technology achieves high EV concentrations for analyses, and improved accuracy for biomarker identification. We have successfully verified the functionality of our technology in a series of in vitro EV isolation and purification experiments, and received an NSERC Idea-to-Innovation (I2I) grant to advance this technology toward commercialization. Here, we are developing this technology for next-generation non-invasive prostate cancer (PCa) active surveillance (AS). We aim to use the technique to understand if there are RNA biomarkers within PCa EVs related to tumor evolution. We will isolate prostate EVs from longitudinally collected plasmas of patients who were previously on AS. These patients elected to defer their prostatectomy, and instead, undergo annual needle biopsies and multiparametric magnetic resonance imaging (mpMRI) to determine if and when their cancer becomes aggressive. We will use our technology to isolate PCa EVs from these samples, submit EVs to RNA isolation and RNAseq, and perform bioinformatics analyses to determine if there are RNA biomarkers correlated to a negative (upgrading) or stable (no upgrading) trajectory. These findings will help bring EV-based cancer monitoring to the clinical setting.