The objective of our projects is to satisfy the need for novel disease biomarkers that reliably predict clinical benefit in tumor recurrence in prostate cancer (PCa). We will utilize multiple different approaches to discover and evaluate novel noninvasive diagnostic and prognostic biomarkers, including measurements of four kallikrein markers and microseminoprotein-beta (MSP) in blood adjusted with the interaction from GWAS-based genetic variation in certain biomarker-encoding gene loci to further improve the specificity in our previously reported detection of lethal PCa-forms at very early, curable stages. We will also use internally standardized qRT-PCR assays to measure circulating tumor cells (CTCs) associated transcripts in whole blood, and use novel ultrasonic wave based microfluidics (acoustophoresis) to enable unbiased, label-free isolation of CTCs and extracellular vesicles (EVs) for subsequent molecular interrogation of the isolated/enriched populations of CTCs and EVs. As our acoustic separation methods are label-free, we are likely to detect new populations of CTCs and EVs compared to currently available methods, particularly due to mesenchyme-epithelium transitioning of CTCs. We aim to characterize the isolated CTCs and EVs after acoustic isolation and identify markers that may enable personalized medicine and more efficient cancer treatment.