The number of new products such as drugs, cosmetics and food additives introduced into the market is growing and poses a significant challenge for risk prediction. Currently, vertebrate testing is the industry standard for hazard assessment but has limitations including high costs, and the growing public and corporate sentiment against using vertebrates for chemical testing. Importantly, current toxicity assessment does not incorporate genetic diversity leading to failure of drug candidates and unintended risks to human sub-populations. The celebrated invertebrate model C. elegans has the potential to fill this gap since thousands of genetically diverse nematode populations are available. However, the main challenge is that chronic studies of chemicals in the nematode model is still tedious, precluding large-scale toxicity testing in nematode populations. This proposal aims to develop a high-throughput technology for chronic toxicity screening of diverse genetic populations in adult C. elegans. The focus of our Phase I plan is to pursue chronic toxicity studies on select toxins and generate foundational data highlighting the impact of genetic diversity, and subsequently scale the NemaLife technology to enable large-scale genetic diversity screening.
AIM 1 : To generate chronic dose-response data on toxins affecting diverse C. elegans strains. We will test the effects of five toxins for which we know the variability in human and rodent responses. Animals will be exposed to toxins daily, followed by scoring of survival, reproductive and neuromuscular health. About 1200 whole-life assays will be conducted with data acquired on 500+ animals per assay condition. We will generate data that will define the rank-order of C. elegans toxin response vis--vis mammalian systems, optimize phenotyping conditions for large numbers of strains per species, and define the amount of heritable variation in toxin 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 the large number of toxins and vast library of genetic strains and wild isolates available, there is a significant need to increase the throughput of our technology. Therefore, we will develop the technology infrastructure and workflow to scale the throughput which includes multiplexing our approach, scale-up of chip fabrication and streamlining data analysis. Technology development at the scale of thousands of whole-life toxicity assays in a few weeks with readouts on animal death and neuromuscular impairments will be a major breakthrough in the field opening up new business opportunities in toxicity testing in the drug and consumer industry.
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.
