Exploiting protein stability changes to identify next generation chemical probes and target engagement technologies

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Background  My research focusses on cutting-edge chemical biology and medicinal chemistry projects that prioritize the concept of "target engagement" - the direct physical interaction of chemical probes with their cellular target. Major aims within this program include developing innovative target engagement technologies and using them to study interactions between chemical probes and their target proteins. Additionally, this program includes an innovative new approach to produce targeted protein degraders (proteolysis targeting chimeras, PROTACs). Objectives and Approach Long-term goal: My research program aims to produce target engagement technologies and PROTACs that are used to explore complex biological phenomena. These tools will allow us to develop a better understanding of our target proteins, such as STAT1, STAT3, CNOT3 and TK1. Further, our novel target engagement technologies will have far-reaching effects within chemical probe discovery, facilitating the development of future generations of chemical probes. Short-term Objectives: 1.Optimize protocols for producing target engagement biosensors within our CeTEAM platform My research team has generated a novel target engagement technology called "CeTEAM" - Cellular Target Engagement by Accumulation of Mutant. This technique uses a biosensor approach for target engagement that is high-throughput and does not require an external stimulus, such as heat. We have demonstrated that CeTEAM biosensors can effectively indicate if an endogenous target protein is engaged by a chemical probe. Within this objective we will expand our repertoire of CeTEAM biosensors and focus on optimizing our process for CeTEAM biosensor development. 2.Develop novel target engagement assays that exploit unique aspects of E. coli biology To improve throughput and reduce barriers to implementation, we have turned to non-mammalian systems for investigating cellular target engagement. In this objective we aim to exploit unique aspects of E. coli biology to produce a novel target engagement technique called LIBR-TEA (ligand-induced Inclusion Body Rescue - Target Engagement Assay, "liberty"). 3. Exploit small biomolecules to produce targeted degraders as chemical biology tools As a highly innovative approach to PROTACs, we will employ naturally occurring biomolecules to target our PROTACs to proteins of interest. As a proof of concept study, we have developed thymidine-inspired PROTACs to target thymidine kinase 1 (TK1) and demonstrated that they can reduce TK1 protein levels in cells. This objective will focus on optimizing bio-inspired PROTACs for TK1 and other targets.  Impact Our novel target engagement techniques and new approach to PROTAC development is set to have a transformative impact on the chemical biology field. These tools will allow us to develop a better understanding of complex biological systems and will facilitate the discovery and validation of future chemical probes and targeted degraders.