Investigating the effects of novel AKT inhibitors on the insulin/IGF-1 like signalling pathway in C. elegans

Caenorhabditis elegans, a free-living nematode, has developed into an exciting model for the study of ageing. It has a lifespan of around three weeks and a number of evolutionarily conserved pathways that can be manipulated to extend its lifespan and healthspan(definition). The best characterised of these pathways is the insulin/IGF-1 signalling pathway (IIS) with the FoxO transcription factor DAF-16 acting as the main target of the pathway. Inhibition of IIS results in the translocation of DAF-16 into the nucleus, allowing it to activate a host of target genes causing lifespan-extension and stress resistance to a wide range of different factors. The aim of this study was to pharmacologically inhibit the IIS pathway using the ATP-competitive AKT inhibitors AZD5363 and AT14148, the allosteric AKT inhibitor MK-2206, the IGF1R inhibitor OSI-906 and the ROCK inhibitor GSK-269962. A C. elegans transgene strain possessing a DAF-16::GFP reporter in order to monitor localisation of DAF-16::GFP under a fluorescent microscope. Worms were treated with the compounds and scored for the nuclear localisation of DAF-16. A bus-8 strain of C. elegans with increased sensitivity to drugs was also used in the investigation. Preliminary findings suggest that MK-2206, AT1148 and GSK-269962 could be having an effect on DAF-16 nuclear localisation. Further studies are needed in order to clarify whether this nuclear localisation is due to inhibition of the IIS pathway or whether it is part of a stress response to high concentrations of the drug.