Electrophilic Metabolite/Drug-Target Discovery and Investigations into On-target Signaling Mechanisms Leveraging Larval Zebrafish and Cultured Cells

This thesis presents novel methodologies and discoveries in the study of reactive electrophilic metabolites and electrophilic drugs, using mammalian cells and live zebrafish larvae as models. Chapter 1 details our current understanding and strategies for studying mitochondrial-derived reactive electrophilic metabolites (mt-REMs). Chapter 2 introduces Z-REX, an innovative platform for delivering specific electrophiles to specific proteins in specific tissues of larval zebrafish, enabling the precise study of ligand/drug-target interactions and associated downstream pathways in live animals with high spatiotemporal resolution and minimal off-target effects. Chapter 3 demonstrates that an immunomodulatory electrophilic drug, dimethyl fumarate (DMF, a derivative of the mt-REM fumarate), elicits an innate immune-cell-specific apoptotic program mediated by a novel KEAP1/WDR1 axis, independent of canonical KEAP1/NRF2 signaling.