A Large-Scale Microfluidics Imaging Platform for High-Throughput Toxicity Testing using Canine Intestinal Organoids

Project SummaryCurrentlytoxicology studies require a large number of animal testing to identify better and safer drug candidatesIn an attempt to reduce the use of large animals in pharmaceutical industrythere is a need to bridge the gap between the conventional in vitro cell based assay and the non human animal studiesRecentlythreedimensional models are gaining momentum and replacing conventionalD cell culture assayD organoids derived from animals can serve the purpose of drug testing recapitulating the drug effect in the whole animalOrganoids derived from dogwith many shared disease pathophysiologyis advantageous since multiple biopsy samples can be operated without sacrificing the animalallowing longitudinal observation of the same animal under the disease pathologyDog spontaneously develops cancer with similar clinical pathology appearancetherapeutic responsesand acquired resistance to treatmentssimilar to that in humanGrowing and investigatingD organoids in large quantities from fresh and frozen biopsy samples are pivotal roles in implementing the large scale screening approach using the organoid based approachIn order to image a large number of organoids for high throughput phenotypic screenswe will pursue our goals in two specific aimswe will develop the microfluidic device and characterize the culturingandperform dose dependent toxicity test by treating the enteroids and coloinds with doxorubicinan intravenousIVchemotherapy agentIn Aimwe will design a new microfluidic platform to immobilize and image ten organoids from eight different populations using a confocal microscopeThe specific geometry of the chip will immobilize the organoids completely in an optically favorable orientation while eliminating motion blur during stage motion to move the samplesIn Aimwe will characterize the growth and physiological states of the two organoidsenterdoids and colonoidsusing multiple passages from both fresh and frozen tissue samplesOn the successful characterization of the cultures and the imaging methodthe organoids will be evaluated for dose toxicity in Aimby treating with Doxorubicinewhich causes gastrointestinal toxicitymucositisin patientsusing multiple stainingThe dose responses will be measured and analyzed using the MCP Mod statisticalbased method and compared to toxicity effects measure on human organoidsin Aimto demonstrate the clinical relevance and the predictive nature of our approachA streamlined protocol and chip fabrication pipeline for large scale production will follow the development of a fully automated large scale organoid screening platformIn Phase IIwe will expand the chip toandwell format for high throughput and develop image analysis algorithm to handle a large volume of images generated from such phenotypic screensLeveraging our unique expertiseNewormics will be able to sale OrganoidChipsTM to the consumer market and simultaneously have the organoid biobank to provide services with the large collection of canine based organotypic models from multiple genetic backgrounds Project NarrativeThe lengthy and escalating price of the drug development process suffers from nearlyfailure ratesa large portion of which is contributed to toxicity and efficacy of new drug moleculesCurrently withdrugs in developmentfaster and better models are needed to screen these compounds to improve the translation and meet the patientsneedespecially for chronic and life threatening disordersOrganotypic models derived from large animals such as dogs recapitulates the human disease pathology better and can be screened in large quantities to identify most effective drug with lowest toxicology effect prior to the first in human test