MEDUSA(GM): an analysis platform for resolving mechanisms of action for anti-cancer drugs - PROJECT SUMMARY/ABSTRACT Understanding mechanisms of drug action is important for identifying settings in which a drug will be effective, and for interpreting – even predicting – potential mechanisms of drug resistance. For anti-cancer drugs, a key aspect of their mechanism of action is the mechanism by which these drugs promote cell death. The first identified death pathway was apoptosis, a form of death that is now reasonably understood. In addition, the community has now identified at least ten distinct forms of regulated necrosis. Our recent studies highlight that these non-apoptotic forms of cell death contribute to the efficacy of many anti-cancer therapies, with each form of non-apoptotic death activated by at least one clinically used compound. A current limitation in the use of drugs that activate non-apoptotic forms of death is that we lack a detailed understanding of how most of these pathways function. To address this issue, we recently developed MEDUSA (Method for Evaluating Death Using a Simulation-assisted Approach). MEDUSA is an analytical method used to evaluate genome-wide CRISPR screens of drug efficacy (which are sometimes referred to as “chemo-genetic profiles”). Our recent publications demonstrate that MEDUSA is uniquely effective at revealing a drug’s mechanism of action and mechanisms of drug-induced lethality, including the signaling and regulatory pathways that facilitate activation of cell death. Furthermore, while many methods exist for evaluating apoptotic death – due to the stability of the apoptotic corpse and the existence of measurable biomarkers that are specific to apoptotic cells – MEDUSA is likely the only method that will be effective at identifying the genetic dependencies and mechanisms of resistance for drugs that activate non-apoptotic forms of cell death. Although MEDUSA is effective, it requires: 1) a detailed understanding of drug function to parameterize a drug-specific simulation, 2) specialized experimental requirements, such as time-resolved data, which are not common in most experiments, and 3) programming proficiency to run the MEDUSA simulations. The central goal of this proposal is to extend the MEDUSA method, to enable easier use and broad utility, particularly in the evaluation of publicly available data. We call our new version MEDUSA: Generalized Method (MEDUSA(GM)). This new method: 1) can be used without any drug-specific information, 2) does not require any specialized data, and 3) will be available on a web portal and can be used without any programming. In this proposal, we will add new functions to our MEDUSA(GM) method, stress test the MEDUSA(GM) method with therapeutically relevant drug scenarios that have not been previously evaluated, and develop the MEDUSA(GM) web portal, which will enable open access use of MEDUSA(GM).
