PRO-FitS: a novel phenotypic assay to identify enhancers of proteostasis in C. elegans

Abstract

The prevalence of neurodegenerative diseases (NDs) continues to rise with the aging of populations worldwide, representing a pressing need for the establishment of therapeutic strategies. Maintaining proteostasis is crucial for healthy aging, as the accumulation of misfolded and aggregated proteins is a key contributor to age-related cellular dysfunction and disease. This study introduces a novel phenotypic assay using Caenorhabditis elegans to screen for small molecule enhancers of proteostasis, aiming at mitigating the proteotoxic stress associated with NDs. This new methodology- PRO-FitS- uses C. elegans motor activity as a proxy for the PROteome Fitness State upon a noxious protein-denaturating stimulus, while allowing a fast and experimenter-free readout. We demonstrate the efficacy of the assay by validating the role of pharmacological mTOR inhibition and serotonergic signaling activation in reducing heat shock-induced proteotoxic damage at the whole-organism level. PRO-FitS will allow the identification of novel compounds that alleviate protein aggregation disorders, potentially revealing new pathways and cellular targets not previously implicated in proteotoxicity.