What Chemical compounds can perturb biological systems - cells or organisms - and the precise manner, in which they do is referred to as their mode-of-action (MoA). MoA-studies are often rate-limiting in the development of new bioactive compounds that could e.g. find use as molecular tools in cellular biology. We will establish an experimental method - called morphological profiling - for making "fingerprints" of bioactive compounds that can be used to gain MoA-information. For instance, similarity between the profiles of an unknown and a well-studied compound indicates that they impact the same cellular processes. Conversely, a unique profile indicates a unique MoA. When scaled to large databases of profiles, this method enables MoA-hypotheses to be generated for any new bioactive compound. Why Alternative profiling methods have previously been used to gain MoA-information, but the transformative aspect of morphological profiling is its high speed and low cost. Whereas MoA-studies previously were only prioritized for a few or even just one bioactive compound within a family, we will now be able to test all analogs as well synthetic intermediates that will e.g. be made during efforts to chemically construct a molecule of interest. In other words, we will be searching for new biological activity (new MoA's) in places that would otherwise never be considered and this potential for discoveries is a very important aspect. Two interesting molecular families will be studied as part of this project, but the perspectives reach much further. How The project will commence with the purchase of a high-throughput microscope needed to establish the morphological profiling method. We will then prepare a series of structural analogs of two different bioactive compounds, one of which is quite toxic to cancer cells while the other is known to stimulate the growth of nerve cells, but common to them is a lack of knowledge about their specific MoA. Morphological profiling of these collections of compounds will allow us to master the method and to learn how the changes in structure impacts the profile and thus alters the cellular activity. We will then combine morphological profiling with other methods to conclusively delineate the MoA for these two compounds. SSR The project involves absolutely cutting-edge methods within studies of bioactive molecules and therefore will be an excellent training ground for the scientists that will be involved. The unique skill set - a mixture of chemistry and cellular biology - that they will acquire on this project will benefit eventual future industrial employers. As mentioned, a key intention of the project is to search for new biological activity and - depending on the context - the proprietary knowledge thus generated can be further explored for its potential commercial potential. Furthermore, we will be able to build a platform that will facilitate collaborations locally as well as with other Danish universities and these interactions will further increase the potential for innovations.