DEVELOPMENT OF A RAPID IN VITRO SCREENING ASSAY FOR DETECTION OF CANCER BIOMARKERS IN URINE AND BLOOD.

The goal of the project is to develop a rapid in vitro screening assay for detection of cancer biomarkers in urine and blood. The aim is testing and optimization of nanoparticle sensors, based on cyanine dyes that are chemically linked to Fe/Fe3O4 nanoparticles via protease-selective consensus (cleavage) sequences. The focus is on the quantitative determination of active cancer-specific proteases in urine and blood via simple fluorescence measurements. The severing from the nanoparticle stops the efficientfluorescence quenching of the organic luminophore by the nanoparticle, thus allowing the detection of the protease activity. The matrix metalloproteinases, urokinase-type plasminogen activator, and cathepsins, are up-regulated in the vast majority of progressing cancers and can, therefore, serve as markers for cell survival/tumor progression, angiogenesis, and tissue remodeling/ invasion. The focus will be on pancreatic cancer, and the studies will be done on urine and blood from 30 patients and 12 healthy individuals. Measuring twelve proteases in total that are linked to pancreatic cancer progression will allow us to obtain statistically significant measurements within 1 hour.