Development and Characterization of a Human Intestinal Organoid Monolayer Model to Predict Oral Drug Disposition

Abstract

The intestine plays an important role in governing the disposition of orally administered medication, serving important gate-keeping functions that can profoundly impact the systemic blood exposure of such medications. While currently used in vitro systems have substantially enhanced our knowledge of intestinal physiology as it relates to drug disposition, they suffer from limitations which often hinders their ability to broadly and accurately predict the fate of orally administered drugs. The use of complex in vitro systems, such as the intestinal microphysiological system (MPS) and adult human stem cell-derived intestinal organoids (enteroids), have profoundly expanded our understanding of intestinal development, physiology, and pathology. However, research employing these models for use in the field of pharmaceutical sciences remains relatively sparse. The projects described in this dissertation proposal seek to develop and characterize an intestinal MPS and a long-term cultured enteroid monolayer model for use in pharmaceutical sciences research.Chapter 2 describes the development and characterization of an intestinal MPS model cultured with LS180 cells, an immortalized colorectal adenocarcinoma cell line, and primary human umbilical vein endothelial cells (HUVECs). In Chapter 3, the development and biochemical characterization of a long-term cultured human enteroid monolayer model was explored. Chapter 4 provides further characterization of the long-term cultured enteroid monolayer model, utilizing RNA-seq deconvolution methods to determine the cell type proportions present in our differentiated enteroid cultures. In Chapter 5, the utility of the enteroid monolayer model to recapitulate and uncover the mechanisms precipitating a select natural product-drug interaction between goldenseal and metformin is explored. This work presented in this dissertation suggests that human enteroid monolayers are a promising model to predict orally administered drug disposition for use in preclinical drug development. Additionally, human enteroid monolayers display promise as a screening tool to predict drug-drug and natural product-drug interactions in vitro to better inform prescribing clinicians and improve the health and safety of patients.