The number of new products such as drugs, cosmetics and food additives introduced into the market is growingand poses a significant challenge for risk prediction. Currently, vertebrate testing is the industry standard forhazard assessment but has limitations including high costs, and the growing public and corporate sentimentagainst using vertebrates for chemical testing. Importantly, current toxicity assessment does not incorporategenetic diversity leading to failure of drug candidates and unintended risks to human sub-populations. Thecelebrated invertebrate model C. elegans has the potential to fill this gap since thousands of genetically diversenematode populations are available. However, the main challenge is that chronic studies of chemicals in thenematode model is still tedious, precluding large-scale toxicity testing in nematode populations. This proposalaims to develop a high-throughput technology for chronic toxicity screening of diverse genetic populations inadult C. elegans. The focus of our Phase I plan is to pursue chronic toxicity studies on select toxins and generatefoundational data highlighting the impact of genetic diversity, and subsequently scale the NemaLife technologyto enable large-scale genetic diversity screening.AIM 1: To generate chronic dose-response data on toxins affecting diverse C. elegans strains. We willtest the effects of five toxins for which we know the variability in human and rodent responses. Animals will beexposed to toxins daily, followed by scoring of survival, reproductive and neuromuscular health. About 1200whole-life assays will be conducted with data acquired on 500+ animals per assay condition. We will generatedata that will define the rank-order of C. elegans toxin response vis-Ã -vis mammalian systems, optimizephenotyping conditions for large numbers of strains per species, and define the amount of heritable variation intoxin responses. These pilot data will validate the capabilities afforded by our screening platform.AIM 2: To scale the throughput of NemaLife Technology for thousands of chronic toxicity assays.Currently, NemaLife's technology has a throughput of hundreds of chronic toxicity assays. However, given thelarge number of toxins and vast library of genetic strains and wild isolates available, there is a significant needto increase the throughput of our technology. Therefore, we will develop the technology infrastructure andworkflow to scale the throughput which includes multiplexing our approach, scale-up of chip fabrication andstreamlining data analysis. Technology development at the scale of thousands of whole-life toxicity assays in afew weeks with readouts on animal death and neuromuscular impairments will be a major breakthrough in thefield opening up new business opportunities in toxicity testing in the drug and consumer industry.
Public Health Relevance Statement: Project Narrative
Non-parasitic nematodes offer a powerful in vivo model to scale and measure toxin responses in
genetically diverse species anchoring a transformative means to bridge current gaps in predictive
toxicology with clinically-relevant end points. Our study aims to combine state-of-the art advances in
microfluidics, computer vision, laboratory automation and a massively curated genetically diverse
nematode species to produce a new screening approach capable of testing thousands of chemicals
relevant to agriculture, nutraceutical and biotech markets.
Project Terms: Adult ; 21+ years old ; Adult Human ; adulthood ; Affect ; Agriculture ; agricultural ; Animals ; Arsenic ; Automation ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Biotechnology ; Biotech ; Cadmium ; Cd element ; Cells ; Cell Body ; Clinical Trials ; Computer Vision Systems ; computer vision ; Cosmetics ; cosmetic product ; Data Analyses ; Data Analysis ; data interpretation ; Cessation of life ; Death ; Pharmaceutical Preparations ; Drugs ; Medication ; Pharmaceutic Preparations ; drug/agent ; Environment ; Food Additives ; Foundations ; Gait ; Gene Library ; genetic library ; Population Genetics ; Genome ; Health ; Human ; Modern Man ; Industry ; Invertebrates ; Invertebrata ; Laboratories ; Life Cycle Stages ; Life Cycle ; life course ; Maintenance ; Metformin ; Dimethylguanylguanidine ; N,N-dimethyl-imidodicarbonimidic diamide ; Dimethylbiguanidine ; Methods ; Genetic Models ; Nematoda ; Nematodes ; roundworm ; Paraquat ; 1,1'-dimethyl-4,4'-bipyridinium ; Methyl Viologen ; Phenotype ; Public Health ; Risk ; Rodent ; Rodentia ; Rodents Mammals ; Safety ; Technology ; Testing ; Toxic Environmental Substances ; Environmental Toxin ; Toxic Environmental Agents ; environmental toxicant ; Toxicology ; Toxin ; Genetic Variation ; Genetic Diversity ; Vertebrates ; Vertebrate Animals ; vertebrata ; Work ; Measures ; Businesses ; Population Heterogeneity ; diverse populations ; heterogeneous population ; population diversity ; Caenorhabditis elegans ; C elegans ; C. elegans ; C.elegans ; base ; Chronic ; Phase ; Variant ; Variation ; Chemicals ; Predisposition ; Susceptibility ; Failure ; Toxicity Tests ; Toxicity Testing ; fluid ; liquid ; Liquid substance ; Exposure to ; Life ; System ; genetic strain ; tech development ; technology development ; Toxicities ; Toxic effect ; novel technologies ; new technology ; reproductive ; Modeling ; response ; high throughput technology ; genetic resource ; µfluidic ; Microfluidics ; Dose ; Data ; in vivo Model ; Nutraceutical ; Molecular ; Pathway interactions ; pathway ; neuromuscular ; Hazard Assessment ; cost ; scale up ; Population ; Heritability ; Impairment ; clinically relevant ; clinical relevance ; resistant strain ; resistance strain ; drug candidate ; screening ; Industry Standard ; Infrastructure ; risk prediction ; forecasting risk ;
