High throughput multiplexed assay for chemicals affecting neuron differentiation

DESCRIPTION provided by applicant High throughput multiplexed assay for chemicals affecting neuron differentiation Neurodevelopmental disorders NDDs affect to of the million children born in the US each year The incidence rates for NDDs have been rising Exposures to toxic chemicals may have contributed to NDDs Due to neuronal development and maturation processes fetus newborns and young children are especially vulnerable to neurotoxins at concentration levels that would not impact adult human health In addition children with genetic susceptibilities to NDDs may have higher risk than non susceptible children For neurodevelopmental toxicity EPA and National Toxicity Program at the National Institute of Environmental Health Science rely on animal studies which are inadequate for subtle neuronal changes not human lengthy in time prohibitively costly with low throughput To address these limitations and to enable high throughput assessment of chemicalsandapos potential impact on human neuronal development we propose an innovative gene expression profiling assay based on differentiating human induced pluripotent stem cells iPSCs iPSCs can be generated from people of various ethnicity and of diverse genetic background which include those who are predisposed to neurodevelopmental and neurodegenerative diseases JuvoBio Pharmaceuticalsandapos scientific team consists of world leading experts in using iPSCs derived neurons to study brain development and disease mechanisms We can reproducibly differentiate iPSCs into high purity neuronal subtypes that are effected by ASD and other NDDs FGNs andgt purity GABAergic neuron andgt and hippocampal neurons andgt in large enough quantities to support high throughput screen By quantitatively assessing the expression level of key markers we can identify the chemicalsandapos impact on neuron differentiation We will ascertain assay conditions that would allow the determination of increased susceptibility of genetically predisposed individual to chemical toxins Our aims Aim Design probes and optimize compound treatment conditions Using six known toxins as reference we will optimize concentration titration timing and length of treatment Aim Define the readout of the assay to predict impact on neuron differentiation The impact of the neurotoxins on quantitative expression of up to differentiation markers for both normal and autism iPSC lines will be analyzed for a smaller set of potentially predictive markers Aim Validate the readout using three alternative assays Readout will be validated using Q PCR antibody staining and mini brain organoid assays PUBLIC HEALTH RELEVANCE There is growing evidence that exposures to toxic chemicals contribute to the growing incidence of neurodevelopment disorders which is a leading and increasingly urgent public health problem in the US and worldwide JuvoBio Pharmaceuticals has developed high throughput platform to differentiate human induced pluripotent stem cells into neurons We propose to use this platform to assess chemicalsandapos potential toxicity on neuron differentiation and to assess if people who are genetically predisposed to neurodevelopment disorders such as autism will have increased risk to chemicalsandapos toxicity