“Targeted Protein Relocalization: Tool Development, Principle Discovery, and Disease Applications”
Subcellular protein localization is crucial for protein function and its disruption is a common feature of cancers and neurodegenerative diseases. The ability to relocalize proteins in a targeted and modular fashion opens new avenues for therapeutic intervention in mislocalization-driven diseases and enables the systematic exploration of how spatial redistribution alters cellular behavior. We develop Targeted Relocalization Activating Molecules (TRAMs) that enable precise modulation of target protein localization via coupling with a shuttle protein. Combined with a custom computational image analysis pipeline, we uncover the hierarchy of protein localization through various TRAM-induced target and shuttle protein combinations. We showcase the ability of TRAM-induced relocalization of NMNAT1 to mitigate degeneration following axonal injury in primary neurons. TRAMs that engage nuclear hormone receptors as shuttles induce nuclear import of mislocalized mutant proteins associated with cancer (SMARCB1Q318X) and neurodegeneration (FUSR495X, TDP43ΔNLS). We demonstrate that this approach works in native physiological contexts, utilizing endogenous nuclear and cytoplasmic shuttles to build an endogenous comparative matrix. Combined, we present TRAM-induced relocalization as a generalizable platform that enables both the discovery of fundamental rules governing protein localization, and the development of novel therapeutic strategies via targeted modulation of subcellular protein distribution.