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Principal Investigator: Sudip Mondal
Organization: VIVOVERSE, LLC
Fiscal Year: 2024
Award: $287,021
Funding agency: National Institute of Environmental Health Sciences
Neurodevelopmental disorders such as ADHD, autism, and learning disabilities are rising globally. In the US, 1
in 6 children suffer from some form of developmental disability. One potentially important factor in their increase
is exposure to chemicals of unknown toxicity in the environment. While there are over 350,000 chemicals in
commercial use and around 13,000 produced in large amounts, only a small percentage have known
toxicological data. Current testing methods for developmental neurotoxicology (DNT) are mostly based on
observing behavioral, physiological, and anatomical defects in small mammals. These studies are prohibitively
time-consuming and expensive for testing the huge backlog of commercial chemicals, and may produce hard-
to-interpret results due to the complexity of observed endpoints and differences between multiple laboratories.
Additionally, many jurisdictions are limiting the numbers of animals that can be used for testing. New approach
methodologies (NAMs) are a new paradigm in toxicology testing that bring together alternative approaches such
as in silico predictions, cell culture, and microphysiological systems-based assays, plus alternative model
animals such as zebrafish, drosophila, and C. elegans. While in silico and in vitro methods can provide high
throughput testing at much lower costs than mammalian studies with easier to interpret and predictive results, in
vitro methods still have substantial costs and complex culturing conditions and neither approach can recapitulate
the effects of a substance on a complete developing organism. C. elegans is an ideal alternative animal model
for DNT testing. It has a complete nervous system with a fully characterized connectome and neuronal lineages,
most of the same important neurotransmitter systems as humans, is fast and easy to grow to maturity in large
numbers, and is amenable to targeted genetic manipulation. We propose to use C. elegans in a comprehensive
DNT test battery where we will track neuronal, synaptic, and behavioral changes as the organism develops from
late larval to adult stage after early exposure to DNT toxicants. In Aim 1, we will develop novel reporter strains
with fluorescent markers for GABAergic and dopaminergic neurons and synapses, expose them to reference
chemicals, and monitor the behavior of growing animals in culture before imaging them on our high-content
microfluidic imaging platform at L4 and D1 stages. Specific behavioral endpoints will be analyzed by ML-based
approaches from the behavioral videos. The high-resolution whole-body 3D images from on-chip imaging will be
analyzed by semi-automated methods for key DNT indicators such as neuronal loss and mislocalization,
membrane blebbing, and defective or missing synapses. All these endpoints will be combined to find the dose
of a chemical at which the point of departure from the control occurs. In Aim 2, we plan to investigate potential
modes of action using expression analysis of developmental and stress genes. We will test a representative
subset of chemicals from the DNT-DIVERS library to establish confidence in our model and lay the foundations
for a fully automated low-cost, high-throughput DNT testing method to assess the risks of common chemicals.
Terms: <0-11 years old><21+ years old><3-D Images><3-D image><3D image><3D images><AD/HD><ADHD><ASD><Adult><Adult Human><Adverse effects><Affect><Age><Anatomic Sites><Anatomic structures><Anatomy><Animal Alternatives><Animal Model><Animal Models and Related Studies><Animal Testing Alternatives><Animals><Assay><Attention deficit hyperactivity disorder><Autism><Autistic Disorder><Behavior><Behavior monitoring><Behavioral><Bioassay><Biological Assay><Biological Markers><Bleb><Blister><Brachydanio rerio><Brain><Brain Nervous System><Bulla><Bullous Lesion><C elegans><C. elegans><C.elegans><Caenorhabditis elegans><Categories><Cell Body><Cell Culture Techniques><Cells><Chemical Exposure><Chemicals><Child><Child Development Disorders><Child Youth><Children (0-21)><Code><Coding System><Color><Common Rat Strains><Complex><Consumption><DA Neuron><Danio rerio><Data><Defect><Development><Developmental Disabilities><Devices><Diagnosis><Dopamine neuron><Dose><Drosophila><Drosophila genus><Drugs><Early Infantile Autism><Encephalon><Environment><Exposure to><Foundations><Future><Gel><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profiling><Genes><Genetic Markers><Hazard Assessment><Human><Image><Immobilization><In Vitro><Individual><Industrialization><Infantile Autism><Intoxication><Kanner's Syndrome><Kinetics><Label><Laboratories><Learning Disabilities><Learning disability><Libraries><Life Cycle><Life Cycle Stages><Mammalia><Mammals><Manuals><Medication><Membrane><Methodology><Methods><Mice><Mice Mammals><Microfluidics><Modeling><Modern Man><Motility><Murine><Mus><NOAEL><Nematoda><Nematodes><Nerve Cells><Nerve Degeneration><Nerve Transmitter Substances><Nerve Unit><Nervous System><Neural Cell><Neural Development><Neurocyte><Neurodevelopmental Disorder><Neurologic Body System><Neurologic Organ System><Neurological Development Disorder><Neuron Degeneration><Neurons><Neurotoxins><Neurotransmitters><No-Observed-Adverse-Effect Level><Organism><Pathway interactions><Pesticides><Pharmaceutical Preparations><Phase><Phenotype><Physiologic><Physiological><Predominantly Hyperactive-Impulsive Type Attention-Deficit Disorder><Predominantly Hyperactive-Impulsive Type Hyperactivity Disorder><Proteins><Publications><Rat><Rats Mammals><Rattus><Regulation><Reporter><Resolution><Risk Assessment><Scientific Publication><Stress><Swimming><Synapses><Synaptic><System><Testing><Three-Dimensional Image><Time><Toxic effect><Toxicities><Toxicology><Transcript Expression Analyses><Transcript Expression Analysis><Validation><Vesication><Whole Organism><Work><Zebra Danio><Zebra Fish><Zebrafish><adulthood><ages><alternatives to animals in research><analyze gene expression><autism spectral disorder><autism spectrum disorder><autistic spectrum disorder><automated analysis><automated image analysis><behavior phenotype><behavioral monitoring><behavioral phenotyping><bio-markers><biologic marker><biomarker><biomarker identification><cell culture><cell cultures><connectome><cost><data to train><dataset to train><developmental><developmental disease><developmental disorder><developmental toxicity><dopaminergic neuron><drug/agent><experiment><experimental research><experimental study><experiments><fruit fly><gene biomarker><gene expression analysis><gene expression assay><gene expression biomarker><gene manipulation><gene marker><gene signature biomarker><genetic biomarker><genetic manipulation><genetically manipulate><genetically perturb><hazard><high resolution imaging><identification of biomarkers><identification of new biomarkers><imaging><imaging platform><in silico><in vitro Model><in vivo><kids><life course><living system><mRNA Expression><marker identification><membrane structure><microphysiologic model><microphysiologic platform><microphysiologic system><microphysiology model><microphysiology platform><microphysiology system><model of animal><model organism><nerve cell death><nerve cell loss><nervous system development><neural degeneration><neural network><neurodegeneration><neurodegenerative><neurodevelopment><neurodevelopmental disease><neurological degeneration><neuron cell death><neuron cell loss><neuron death><neuron development><neuron loss><neuron toxicity><neuronal><neuronal cell death><neuronal cell loss><neuronal death><neuronal degeneration><neuronal development><neuronal loss><neuronal toxicity><neurotoxic><neurotoxicant><neurotoxicity><neurotoxicology><new approaches><novel><novel approaches><novel strategies><novel strategy><orthopedic freezing><pathway><reproductive toxicity><research alternatives to animal use><resolutions><roundworm><specific biomarkers><synapse><synapse formation><synaptogenesis><toxicant><training data><transcriptional profiling><validations><welfare><youngster><µfluidic>