Inhibitor-Mediated Programming of Glycoforms

Site-directed mutagenesis revolutionised the study of proteins and enabled the development of protein-based therapeutics. Our long-term vision is to have equivalent impact through the precise manipulation of the glycans (carbohydrates) on cells and recombinant proteins that will enable the discovery and production of the next generation of therapies for cancer, neurodegeneration and other disease families. The glycans that are present on most proteins and cells have a substantial impact on their biological functions, yet the untemplated nature of their synthesis leads to inherent heterogeneity in both their structure and activity. This heterogeneity is very difficult to control, making it impossible to generate defined glycan ensembles with optimal activity using current technology. Here we present a radically new approach to the controlled manipulation of glycans that will be the functional equivalent of site-directed mutagenesis for manipulating proteins. We will use the delivery of computationally defined mixtures of enzyme-specific inhibitors to the site of glycan biosynthesis in the cell to tune the activity of glycosyltransferases. Our approach, termed Inhibitor-Mediated Programming of Glycoforms (IMProGlyco) will provide an effective strategy to manipulate the glycosylation machinery and thereby generate proteins with defined ensembles of glycans. It will enable the production of precision glycan engineered therapeutic proteins and vaccines. Moreover, shaping cellular glycan profiles will aid discovery science to uncover glycan functions and improve therapeutic cells, such as those used in Chimeric Antigen Receptors cell Therapy (CAR-T). Our technology will be adaptable and expandable into other cell types and organisms allowing glycan shaping in all areas of eukaryotic cell biology to enable new biotechnological applications and fundamental studies of biology.