Project Summary/Abstract The goal of this proposal is to investigate the role of a human microphysiologic intestine-on-a-chip platform as a precision medicine tool to model a devastating disease affecting premature infants known as necrotizing enterocolitis (NEC). We developed preclinical models of NEC using both organoids and a “NEC-on-a-chip” model system to recapitulate the intestinal environment of the human disease in vitro, gain new insights into disease pathogenesis and test the functional and clinical utility of our models to evaluate the efficacy of candidate therapeutics. Our NEC-on-a-chip model utilizes a combination of premature infant intestinal organoids along with human endothelial cells and patient-derived microbiota, to recreate critical aspects of premature gut pathophysiology. Our preliminary studies demonstrate that co-culture of these components on intestine-on-a- chip microfluidic devices produces clinical features seen in human NEC such as gut barrier failure with the breakdown of cellular tight junctions, decreased epithelial cell proliferation, a dramatic increase in the pro- inflammatory cytokine response, as well as a significant amount of cell death. In this proposal, we will use several multi-omic approaches to characterize our NEC-on-a-chip model and compare to the human NEC phenotype. To achieve this, we developed a multi-center NEC Biorepository, which consists of detailed clinical metadata corresponding to a plethora of human specimens, including intestinal organoids cultured from the biopsies of premature infants with or without NEC. Furthermore, we have created a high-throughput and high-content drug screening platform using premature intestinal organoids to identify drugs or compounds that inhibit the pathogenic inflammatory responses seen in vitro. Moreover, we will demonstrate the functional and clinical utility of our patient-derived NEC-on-a-chip model as a precision medicine platform to test the dosing, efficacy, and toxicity of candidate therapeutics. To successfully complete these studies, we established a multi-disciplinary team with the expertise of a Neonatologist, Cell Biologist, Pediatric Surgeon, Genome Scientist and Bioinformatician. Taken together, these studies will make a significant conceptual advance in our understanding of the multicellular interactions with the microbiome of the developing premature intestine and provide new model systems and preclinical platforms by which the identification and testing of therapeutics for NEC and other intestinal diseases can be performed in this vulnerable patient population.