MetabolGut: a rapid assay platform to evaluate the impact drugs on lipid-handlingpathways and chylomicron-associated drug distribution using stem cell-drivenhuman absorptive enterocytes.

Project Summary
Maintaining healthy physiology of the human gut is a large focus of Pharma. Models to study
drug/nutrient absorption, xenobiotics, toxicology, and preclinical efficacies are hampered by the lack of
accurate, reproducible, and easy to use cell culture models to evaluate such topics. For these reasons,
there is a strong need for better in vitro models that recapitulate disease states of the human gut, and
better platforms for drug discovery and validation. Lipid-handling is central to human health
conditions and pharmacokinetics. Lipid-related metabolic disorders (i.e. obesity, insulin resistance,
hyperlipidemia, and hypertension) are a global epidemic and predicted to increase as sedentary jobs and
unhealthy diets increase. Treatments for metabolic disorders are sparse with limited efficacy highlighting
the need for more broadly effective drugs. Lipid-handling mechanisms by Absorptive Enterocytes (AEs)
can strongly influence oral drug Absorption, Distribution, Metabolism and Excretion (ADME), and drug
bioavailability can be negatively and positively regulated by AE lipid-handing as many lipid soluble
drugs are associated with chylomicrons (CMs). CM-associated-drug export by AEs is a first-line metric
of bioavailability for lipophilic drugs with no accurate preclinical model. New tactics that harness lipid-
handling mechanisms have strong potential to improve drug engineering for metabolic diseases,
bioavailability and efficacy. To meet this need Altis Biosystems Inc., an early-stage biotechnology
company, will collaborate with scientists at the University of North Carolina at Chapel Hill to develop
Absorptive Enterocytes (AEs) on a high-throughput 96-Traswell format. ‘MetabolGut’ is a monolayer of
differentiated AEs derived from the foundational technology, RepliGutTM, which is a stem cell-driven
monolayer of human epithelium derived the small intestine or colon of organ donors and contains all of
the proliferative and differentiated cell types found in vivo. The goal of this Phase I proposal is to
develop four innovative elements: 1) rapid fluorescent readouts for lipid absorption and export, 2)
simultaneous rapid detection of barrier integrity and fatty acid export, 3) rapid and highly sensitive
identification of fatty acid metabolic species using non-radioisotope methods, 4) rapid fluorescent
quantification of chylomicron (CM) export. While elements 1-3 have shown substantial utility for
academic studies, they will be explored for commercial viability. Element 4 will be developed de novo as
it represents a new assay for high-throughput detection of CM export and CM-associated drug
quantification.