Controlling Epithelial Polarity: A Human Enteroid Model for Host-Pathogen Interactions

Cell Reports
Julia Y CoManuel R Amieva

Abstract

Human enteroids-epithelial spheroids derived from primary gastrointestinal tissue-are a promising model to study pathogen-epithelial interactions. However, accessing the apical enteroid surface is challenging because it is enclosed within the spheroid. We developed a technique to reverse enteroid polarity such that the apical surface everts to face the media. Apical-out enteroids maintain proper polarity and barrier function, differentiate into the major intestinal epithelial cell (IEC) types, and exhibit polarized absorption of nutrients. We used this model to study host-pathogen interactions and identified distinct polarity-specific patterns of infection by invasive enteropathogens. Salmonella enterica serovar Typhimurium targets IEC apical surfaces for invasion via cytoskeletal rearrangements, and Listeria monocytogenes, which binds to basolateral receptors, invade apical surfaces at sites of cell extrusion. Despite different modes of entry, both pathogens exit the epithelium within apically extruding enteroid cells. This model will enable further examination of IECs in health and disease.

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Methods Mentioned

BETA
IEC
biopsies
dissection
flow cytometry
confocal microscopy
Assay
IECs
dissecting
PCR
fluorescence microscopy

Software Mentioned

Zen
Volocity Image Analysis
GraphPad Prism
OpenLab
Improvision
FlowJo
ImageJ

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