A high throughput alcohol screening platform (HT-ASP) incorporating cortical organoids.

ABSTRACT Organoids are spontaneously formed heterogeneous, multi-cellular 3D tissues created by differentiating progenitor cells (often induced pluripotent stem cells, iPSCs). Organoids-based MPS models currently represent the most complex microscale, in vitro models of the human brain and the best in vitro model for drug screening. However, establishing organoid MPS models is complex, requires significant hands-on time for regular medium changes (often every 48 hours or less) and multiple differentiation mediums. Variability in feeding schedule and handling protocols creates further unwanted variability in organoid culture, which is difficult to overcome in a manual system. Moreover, alcohol is a complex insult with many, diverse effects on the human brain. While this makes brain organoids heterogenicity valuable, no single organoid system replicates the entire brain and therefore there is a need to determine how well a specific organoid can respond to the alcohol insult. Moreover, no single brain organoid replicates the entire human brain. Alcohol affects many parts of the brain (Figure 1), including the basal ganglia, extended amygdala, and the prefrontal cortex. While the deep brain structures are heavily involved in intoxication and the negative effects of withdrawal, preoccupation and anticipation, the effects that most often lead to relapse in recovering patients, occur in the prefrontal cortex which is responsible for conscious decision making and executive function. Current, approved medication for the treatment of alcohol use disorder (AUD) focus on this area of the brain. Therefore, we propose that cortical organoids are a useful model for screening potential therapeutics. Overall, we propose to adapt our developed microphysiological system (MPS) platform for the development of vascularized thick tissue and our organoid as a living biological sensor platform to create a high-throughput alcohol assay platform (HT-ASP) that will incorporate automation to mature, maintain, and assay over 100 organoids in culture. The developed platform will provide a highly robust and genetically diverse platform for screening multiple drug candidates.