Bright nanoparticle probes for amplified fluorescence in situ hybridization in cancer diagnostics
Rapidly expanding field of cancer diagnostics generates strong demand for new biosensing tools. In particular, it concerns probes for fluorescence in situ hybridization (FISH), that enables detection of DNA and RNA cancer biomarkers directly in diseased cells. Although FISH technique has already been accepted for cancer diagnostics in clinics, its applications are still limited because the method is slow, expensive and requires complicated amplification protocols to obtain sufficient fluorescence signal. Here, we propose a solution based on ultrabright DNA-functionalized dye-loaded fluorescent polymeric nanoparticles recently developed within the ERC grant BrightSens and protected by two patent applications. We aim to develop ultrabright FISH probes (nanoprobes) for cancer research and clinical diagnostics. These nanoprobes will feature single-molecule DNA/RNA sensitivity, low cost, fast and direct one step cell staining protocol, and compatibility with clinical samples and single-cell assays. The amplification of FISH performance will be ensured by our dye-loaded polymeric nanoparticles of 7-20 nm size that are ~1000 fold brighter than single organic dyes. The project is composed of five tasks: (1) Synthesis and optimization of FISH nanoprobes; (2) validation of FISH nanoprobes in cancer cell lines; (3) development of multiplexing assays for detection at least 9 biomarkers; (4) their validation in clinically relevant samples; and, finally, (5) commercialization of FISH nanoprobes. The developed probes will constitute highly competitive products that can greatly improve performance of FISH assays in clinics and research laboratories and thus impact human health. Therefore, they will be directly proposed for commercialization by licensing to existing companies and/or through creation of a startup.