Tracking the effect of cell senescence on cancer stem cell formation in vitro using a nanoparticle-based reporter system

The project aims to increase our knowledge of cellular ageing and improve its detection. In all experiments with ageing cells, the first step is to obtain them under laboratory conditions. By observing markers of ageing, the formation of these cells can be confirmed. One marker of ageing cells is the observed lack of cell division. Another is a protein called β-galactosidase, which is present in significantly higher amounts in ageing cells than in non-ageing cells. Typically, the presence of these markers can be detected in two separate tests. To improve the efficiency of this process, we will develop a nanotechnology that simultaneously confirms both markers in a single test. Our invention is based on nanoparticles that emit fluorescence when a cell undergoes cellular ageing due to stress or damage. This method will allow the identification of live ageing cells from a population for subsequent quantitative analysis by flow cytometry. The project plan consists of three main tasks involving interdisciplinary research in biology, with links to medicine, chemistry and, to some extent, physics. In the first step, we will synthesise fluorogenic nanoparticles. We will investigate the best methods to obtain them and develop protocols to test their quality. We will then study the biological fate of the nanoparticles inside the target cells. We will investigate the capabilities of the nanoparticle-based reporter system using a variety of cell lines. It is worth noting that our team has recently observed that ageing markers can appear in cancer cells, requiring other methods to confirm their presence. The third aim of the project is to understand the mechanism by which dormant cancer cells (senescent cells) are reprogrammed into disease-causing cancer stem cells (CSCs). The development of our nanoparticle-based reporter system will allow us to understand how the same type of cell in our body behaves and reacts under different conditions. The origin of CSCs is poorly understood. Understanding their nature at the molecular level will allow us to find new methods to identify their source and block their formation. One potential outcome of our concept is its use in biomedical applications, drug discovery and diagnosis of age-related diseases.