High throughput small molecule screening reveals NRF2-dependent and -independent pathways of cellular stress resistance

Biological aging is the dominant risk factor for most chronic diseases. Development of anti-aging interventions offers the promise of simultaneously preventing or even treating many chronic illnesses. Cellular stress resistance is an evolutionarily conserved feature of longevity. Here, we identify compounds that confer resistance to the superoxide generator paraquat (PQ), the heavy metal cadmium (Cd), and the DNA alkylator methyl methanesulfonate (MMS). Some rescue compounds conferred resistance to a single stressor, while others conferred multiplex stress resistance. Induction of stress resistance in mouse fibroblasts was highly predictive of longevity extension in a published large-scale longevity screen in C. elegans. Transcriptomic analysis implicated NRF2 signaling in stress resistance provided by two protective compounds, cardamonin and AEG 3482. Molecules that conferred resistance to PQ also induced cellular inflammatory pathways, and core pathways such as AMPK signaling. Following further optimization, small molecules identified in this work, particularly those that promote resistance to multiple distinct stressors, may represent attractive candidates to evaluate for potential pro-health and pro-longevity effects in mammals. Overall design: RNA-seq was performed on mouse tail fibroblasts treated with a drug (three replicates) or DMSO control (5 replicates), with a total of 30 samples on an Illumina HiSeq4000. The transcriptome was analyzed using Kallisto, counted using Tximport, and differential expression was determined using DESeq2.