SBIR-STTR Award

This project responds to the NIEHS RFA-ES-20-005 “Organotypic culture models developed from experimental animals for chemical toxicity screening,” that is aligned with the needs of the NTP’s Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) for development and evaluation of new, revised, and alternative methods to identify potential hazards to human health and the environment. Through our ongoing NCATS SBIR Phase II, Lena Biosciences (LB) developed and commercialized an organ- on-a-chip-like, Perfused Organ Panel, with a proprietary liquid breathing technology that provides uniform interstitial perfusion, superior delivery of oxygen and stable pH to 48 statistically independent organ cultures. In this project, Lena Biosciences will use the Perfused Organ Panel to develop physiologically-relevant in vitro screening systems using cells derived from animal species typically utilized for toxicological testing. Next, LB will replicate biological interactions and toxicological responses observed in animal tissues. Lastly, LB will produce assay data that is suitable for comparisons between in vitro and in vivo animal toxicology studies, and Tox21 HTS data. Ultimately, this project will provide thoroughly characterized and validated, alternative in vitro test systems with high specificity and sensitivity to reduce or replace the use of animals in toxicity testing. In Phase I, LB will develop biologically and xenobiotic-metabolically competent rodent liver and brain models having in vivo like cellular respiratory metabolism to achieve optimal mitochondrial responsiveness and susceptibility to toxicants. The liver model will provide high activity of drug metabolizing enzymes for in situ generation of reactive metabolites, and mimic parallel processes of parent drug deletion and metabolite formation to better model and predict toxicological outcomes. The brain model(s) will mimic the brain’s innate immunity with robust toxicological responses to drug overdose, Acetylcholinesterase inhibition with acute neurotoxicity (Phase I), and neuropathy target esterase inhibition that cause delayed neuropathy (Phase II) following the exposure to organophosphorus (OP) chemicals and their toxic metabolites. Perfused Organ Panel and 3 sets of in vitro controls will be treated with Acetaminophen (APAP) and Malathion, an OP insecticide with a neurotoxic metabolite, Malaoxon, that are relevant to the testing of specific tissue models and for which species-matched in vivo data already exists. This will facilitate benchmarking and show the Perfused Organ Panel’s utility as an alternative to in vivo models currently used by the U.S. Environmental Protection Agency. To successfully carry out these studies and ensure the project’s success, we assembled a team of experts in advanced cell culture models of liver and brain (LB’s PI and CSO, Dr. Shoemaker), drug metabolism and metabolite formation (Dr. Morgan, Emory, Department of Pharmacology and Chemical Biology), APAP toxicity (Dr. Jaeschke, University of Kansas Medical Center), and brain and liver toxicology (Dr. Caudle, Emory, Department of Environmental Health, Dr. Jaeschke, and Dr. Morgan).

Public Health Relevance Statement:
This project will lay out foundation for new, alternative, animal-relevant test systems for sensitive, reliable and predictive in vitro testing of environmental toxicants, such as organophosphate and carbamate pesticides, to ethically and humanely identify potential hazards to human health and the environment.

Project Terms:
3-Dimensional; Acetaminophen; acetaminophen overdose; Acetylcholinesterase; Ache; Acute; Agreement; Albumins; animal tissue; Animals; Basal metabolic rate; Benchmarking; Biological; Biological Assay; Biological Markers; Biological Models; Biological Sciences; biological systems; Biology; Brain; Breathing; Carbamates; Cell Culture Techniques; Cell Survival; cell type; Cells; Chemical Warfare Agents; Chemicals; Cholinesterase Inhibitors; cost; Data; Database Management Systems; Databases; drug metabolism; Endotoxins; Engineering; Ensure; Environment; Environmental Health; Enzymes; Ethics; Evaluation; experience; Exposure to; Foundations; Funding; Gases; Generations; glucose uptake; Glutathione; Goals; hazard; Health; Hour; Human; improved; In Situ; In Vitro; in vitro Model; in vitro testing; in vivo; in vivo Model; Inflammatory Response; Innate Immune Response; Insecticides; interstitial; Journals; Kansas; Legal patent; Lipid Peroxidation; Liquid substance; Liver; liver injury; Lung; Lymphatic; malaoxon; Malathion; Measures; Medical center; Metabolic; Metabolic Pathway; Metabolism; metabolome; method development; Methods; Mitochondria; Modeling; Monitor; mouse model; N-acetyl-4-benzoquinoneimine; National Institute of Environmental Health Sciences; Natural Immunity; Nervous system structure; Neuropathy; neuropathy target esterase; neurotoxic; neurotoxicity; Nonlytic; novel; Organ; Organ Culture Techniques; organ on a chip; Organophosphates; organophosphorus insecticide; Outcome; Overdose; Oxygen; Parents; Peer Review; Perfusion; Pesticides; Pharmaceutical Preparations; Pharmacology; Phase; Physiological; Positioning Attribute; Predisposition; Process; Publications; Publishing; Reactive Oxygen Species; Research; respiratory; response; Rodent; Sampling; scaffold; screening; Sensitivity and Specificity; Small Business Innovation Research Grant; success; Synapses; System; Technology; Testing; Thiobarbituric Acid Reactive Substances; three dimensional cell culture; Time; Tissue Model; Tissues; tool; Toxic effect; Toxic Environmental Substances; toxicant; Toxicity Tests; Toxicology; United States Environmental Protection Agency; Universities; Urea; Xenobiotic Metabolism; Xenobiotics

This project responds to the NIEHS RFA-ES-20-005 "Organotypic culture models developed from experimentalanimals for chemical toxicity screening," that is aligned with the needs of the NTP's Interagency Center for theEvaluation of Alternative Toxicological Methods (NICEATM) for development and evaluation of new, revised,and alternative methods to identify potential hazards to human health and the environment.Through our ongoing NCATS SBIR Phase II, Lena Biosciences (LB) developed and commercialized an organ-on-a-chip-like, Perfused Organ Panel, with a proprietary liquid breathing technology that provides uniforminterstitial perfusion, superior delivery of oxygen and stable pH to 48 statistically independent organ cultures.In this project, Lena Biosciences will use the Perfused Organ Panel to develop physiologically-relevant in vitroscreening systems using cells derived from animal species typically utilized for toxicological testing. Next, LBwill replicate biological interactions and toxicological responses observed in animal tissues. Lastly, LB willproduce assay data that is suitable for comparisons between in vitro and in vivo animal toxicology studies, andTox21 HTS data. Ultimately, this project will provide thoroughly characterized and validated, alternative in vitrotest systems with high specificity and sensitivity to reduce or replace the use of animals in toxicity testing.In Phase I, LB will develop biologically and xenobiotic-metabolically competent rodent liver and brain modelshaving in vivo like cellular respiratory metabolism to achieve optimal mitochondrial responsiveness andsusceptibility to toxicants. The liver model will provide high activity of drug metabolizing enzymes for in situgeneration of reactive metabolites, and mimic parallel processes of parent drug deletion and metaboliteformation to better model and predict toxicological outcomes. The brain model(s) will mimic the brain's innateimmunity with robust toxicological responses to drug overdose, Acetylcholinesterase inhibition with acuteneurotoxicity (Phase I), and neuropathy target esterase inhibition that cause delayed neuropathy (Phase II)following the exposure to organophosphorus (OP) chemicals and their toxic metabolites.Perfused Organ Panel and 3 sets of in vitro controls will be treated with Acetaminophen (APAP) and Malathion,an OP insecticide with a neurotoxic metabolite, Malaoxon, that are relevant to the testing of specific tissuemodels and for which species-matched in vivo data already exists. This will facilitate benchmarking and showthe Perfused Organ Panel's utility as an alternative to in vivo models currently used by the U.S. EnvironmentalProtection Agency. To successfully carry out these studies and ensure the project's success, we assembled ateam of experts in advanced cell culture models of liver and brain (LB's PI and CSO, Dr. Shoemaker), drugmetabolism and metabolite formation (Dr. Morgan, Emory, Department of Pharmacology and ChemicalBiology), APAP toxicity (Dr. Jaeschke, University of Kansas Medical Center), and brain and liver toxicology (Dr.Caudle, Emory, Department of Environmental Health, Dr. Jaeschke, and Dr. Morgan).

Public Health Relevance Statement:
This project will lay out foundation for new, alternative, animal-relevant test systems for sensitive, reliable and predictive in vitro testing of environmental toxicants, such as organophosphate and carbamate pesticides, to ethically and humanely identify potential hazards to human health and the environment.

Project Terms:
<3-Dimensional><3-D><3D><3-D modeling><3D modeling>