Hydrogel-enabled self-assembled human brain organoids for developmental neurotoxicity applications

Project Summary AbstractThere is a well recognized need for more accurate and cost effective toxicology screening for therapeutic and environmental compoundsCurrent methods using cell cultures or animal models lack predictability and can be costlyRecent advances in the development of stem cell derived brain organoids have led to increasing interest in these models for neural toxicology screening as well as for drug screening and the study of neural developmental mechanismsHoweverbecause of their costcomplexityand workflow requirementsthese methods have yet to be effectively implemented beyond research applicationsCurrently neural organoids are typically formed on a complex extracellular matrix derived from tumors cultured in rodentscommonly known as MatrigelThe process is cumbersomemakes interrogation difficultand lacks reproducibilityMore recentlyinvestigatorsincluding a Stem Pharm cofounderhave demonstrated that complex neural organoids can be formedculturedand assayed reproducibly in a plate based system on engineered hydrogel substrates with human embryonic stemEScell derived precursor cellsWork in this proposal will further that development and lead to the validation of an iPSC derived neural organoid that can be self assembled and cultured on Stem Pharmandapos s specialized synthetic hydrogel in awell plate format that is amenable to automated screening applicationsSpecific Aims willvalidate iPSC derived human neural organoids formed on Stem Pharm synthetic hydrogels utilizing neural progenitors as well as vascular and microglial cellstransition the organoid formationgrowthand interrogation to awell plate formatThis will require experimentation and optimization of hydrogel characteristicscell seeding densitiesand media replenishmentOrganoid sizemorphologyneural maturationand health will be assessed using microscopy and immunologic and molecular methods and will yield potential biomarkers for future studiesandcompare the inflammatory cytokine response to lipopolysaccharideLPSmicroglial activation in the neural organoid system to the response generated by microglia in isolation in adimensionalDculture systemCompletion of this study will result in a hydrogel composition and culturing protocols to support the formation of reproducible brain organoids in medium throughput toxicology screening applicationsPhase II studies will utilize data from Phase I transcript expression analyses to identify quantitative molecular panels to assess toxicity and develop phenotypic screens that will be used to validate these assays with known developmental neural toxins and non toxic controlsThis work will develop more accurate models for neural health and pathogenesis and ultimately lead to commercially available assays for screening applications Project Narrative This project will develop human neural organoid models that are amenable to toxicology screening inwell formatsaddressing the well recognized need for more accurate and costeffective methodsThe study will use Stem Pharmandapos s specialized hydrogel material for the formation of neural organoids from induced pluripotent stem cell derived cellsValidation of this model and development of biomarkers associated with toxicity will lead to commercially available accurate assays for screening chemicals and drug candidates for neurotoxicity