Grant: Discovery Grants Program - Individual

Carbohydrates (also called glycans) play critical roles in many biological processes. Over the past 20 years my lab has developed state-of-the-art mass spectrometry (MS) methods. We apply these methods to accelerate discovery and characterization of biologically-relevant interactions between glycans and glycan-binding proteins (GBPs) and evaluate the activities and substrate specificities of Carbohydrate-Active enZymes (CAZymes). With these methods, we further our understanding of the roles that these glycan interactions play in human health and disease. In this Discovery Grant application, we propose three new projects that push the limits of current MS capabilities for interrogating glycan binding and processing. This research will lead first and foremost to innovative MS methods to identify new glycan-GBP interactions, quantify their thermodynamic properties and measure CAZyme activities under physiological conditions. These methods will also be used in down-stream studies with collaborators to build a deeper appreciation of the biological implications of glycan recognition and processing and to translate these findings into new therapeutics and diagnostics. Our three projects are: 1. Developing a MS-based shotgun glycomics assay for discovery of natural glycan ligands of GBPs. The assay will combine catch-and-release electrospray ionization (CaR-ESI)-MS and time-resolved thermal dissociation measurements. We will implement the assay using a newly acquired 15T Fourier transform ion cyclotron resonance mass spectrometer (the first of its kind in Canada). We will then apply CaR-ESI-MS to identify natural ligands of GBP targets that have high therapeutic value. 2. Developing a MS-based assay to simultaneously identify and quantify GBP binding to mixtures of glycolipids in membranes. Application of this assay will improve molecular-level understanding of cell recognition by GBPs. Such insights will guide the rational design of inhibitors and therapeutic antibodies that target glycolipids. 3. Developing an ESI-MS assay that is compatible with biological fluids, to quantify CAZyme activities. Through collaborations with clinicians we will apply the assay to clinical samples to elucidate how CAZyme activities are affected by specific cancers. These insights have potential in developing a new, non-invasive method to diagnose various cancers. The planned projects will train six graduate students and one undergraduate each year. All trainees will gain skills in high-performance MS and its applications to glycan binding and processing. In some projects, they will gain specialized skills in binding and enzyme assay development, and in immunology and cancer biomarker discovery. They will work in an environment that has been highly successful in training individuals for careers in the natural sciences and engineering, one that combines outstanding research training with professional development opportunities tailored to each individual's career objectives.