Inhibition of the neuromuscular acetylcholine receptor with atracurium activates FOXO/DAF-16-induced longevity Rebecca L. McIntyre1, Simone W. Denis1, Rashmi Kamble1, Marte Molenaars1, Michael Petr2, Bauke V. Schomakers1,3, Mizan Rahman4, Siddhartha Gupta5, Marton Toth5, Siva A. Vanapalli4,5, Aldo Jongejan6, Morten Scheibye-Knudsen2, Riekelt H. Houtkooper1, Georges E. Janssens1* 1 - Laboratory Genetic Metabolic Diseases, Amsterdam Gastroenterology and Metabolism, Amsterdam Cardiovascular Sciences, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands 2 - Center for Healthy Aging, Department of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark 3 - Core Facility Metabolomics, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands 4 - Dept. of Chemical Engineering, Texas Tech University, Lubbock, TX, USA 5 - NemaLife Inc., Lubbock, TX, USA 6 - Bioinformatics Laboratory, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands Transcriptome-based drug screening is emerging as a powerful tool to identify geroprotective compounds to intervene in age-related disease. We hypothesized that, by mimicking the transcriptional signature of the highly conserved longevity intervention of FOXO3 (daf-16 in worms) overexpression, we could identify and repurpose compounds with similar downstream effects to increase longevity. Our in silico screen, utilizing the LINCS transcriptome database of genetic and compound interventions, identified several FDA-approved compounds that activate FOXO downstream targets in mammalian cells. These included the neuromuscular blocker atracurium, which also robustly extends both lifespan and healthspan in C. elegans. This longevity is dependent on both daf-16 signaling and inhibition of the neuromuscular acetylcholine receptor subunit unc-38. Both atracurium treatment and unc-38 RNAi similarly improve healthspan, lifespan, and stimulate DAF-16 nuclear localization. Finally, using RNAseq transcriptomics, we show that atracurium activates DAF-16 downstream effectors. Together, these data demonstrate the capacity to mimic genetic lifespan interventions with drugs, and in doing so, reveal that the neuromuscular acetylcholine receptor regulates the highly conserved FOXO/DAF-16 longevity pathway.