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Using automated methods to quantify both efficacy and toxicity of compounds for drug development using the nematode worm Caenorhabditis elegans
Introduction/Background & aims The nematode Caenorhabditis elegans has been an extremely successful model organism for understanding fundamental biology but is currently not a standard component of the industrial drug discovery process, despite the benefits of using a small, genetically amenable animal model to test both efficacy and toxicity without any regulatory restrictions. One potential barrier to uptake for pharmacological studies is the lack of standardised quantitative assays. Most assays for animal health require manual scoring by a skilled researcher. Here we describe an automated system that provides a data rich readout and straightforward quantification of both developmental and reproductive toxicity and healthspan.
Method/Summary of work Media: Defined amino acid media in agar in 60 mm petri dishes (Maynard et al 2018).
Compounds: FUdR (floxuridine, 5-Fluorodeoxyuridine), sulfamethoxazole (SMX), metformin were added to the agar before pouring into the Petri dishes.
Bacteria: E. coli OP50: an overnight culture in LB was added to the top of the agar and allow to grow for 48 hours at 24°C
Worms: C. elegans N2 for developmental and reproductive toxicity, SS105 glp-4(bn2), strain that is sterile at 24°C was used for healthspan measurements.
Imaging: Healthspan machine multicamera imaging with automated movement tracking and quantification.
Results/Discussion In developmental and reproductive toxicity studies, FUdR (0.5microM) resulted in a 17% increase in the number of days taken to reach a population threshold of 125 progeny/gravid mother (n=8), without any significant delay in the start of egg-laying. In anti-ageing efficacy studies, metformin (12.5, 25 and 50 mM) resulted in an increase in the mean number of moving hours of 18.5, 33 and 48% compared to solvent control, respectively, over 10 days (n= 350). This increase was significant at 25 and 50mM, but not 12.5 mM. Similar studies with sulfamethoxazole (SMX, 1, 4, 16 microg/ml) resulted in a 2.5 % decrease at 1 microg/ml, but 14 and 34% increase at 4 and 16 microg/ml, respectively, over 12.5 days. The increase was significant between day 2 and day 4 for 4 microg/ml and between day 2 and day 7 for 16 microg/ml, but not before or beyond those periods.
Conclusion(s) This method is more sensitive and gives richer data than manual methods. As well as showing efficacy for compounds that prolong health during ageing, the methods can be used to assess acute, developmental, reproductive and chronic toxicity before mouse studies. Once a compound shows positive effects in C. elegans, the C. elegans system can also be used for target identification and drug screens of similar compounds in a chemical series.