Enteroid Monolayers Reveal an Autonomous WNT and BMP Circuit Controlling Intestinal Epithelial Growth and Organization

Abstract

The intestinal epithelium maintains a remarkable balance between proliferation and differentiation despite rapid cellular turnover. A central challenge is to elucidate mechanisms required for robust control of tissue renewal. Opposing WNT and BMP signaling is essential in establishing epithelial homeostasis. However, it has been difficult to disentangle contributions from multiple sources of morphogen signals in the tissue. Here, to dissect epithelial-autonomous morphogenic signaling circuits, we developed an enteroid monolayer culture system that recapitulates four key properties of the intestinal epithelium, namely the ability to maintain proliferative and differentiated zones, self-renew, polarize, and generate major intestinal cell types. We systematically perturb intrinsic and extrinsic sources of WNT and BMP signals to reveal a core morphogenic circuit that controls proliferation, tissue organization, and cell fate. Our work demonstrates the ability of intestinal epithelium, even in the absence of 3D tissue architecture, to control its own growth and organization through morphogen-mediated feedback.

Regulatory network analysis of Paneth cell and goblet cell enriched gut organoids using transcriptomics approaches

“…This finding correlates with previous investigations at both the transcriptomic and proteomic levels (25,(27)(28)(29) and is likely due to the shared differentiation pathways of these secretory cells. Nevertheless, as enteroids contain a mixed population of cell types by nature and because intercellular communication is key to a functioning epithelium (19,71), the increased proportion of non-targeted secretory lineages should not be an issue for the application of these models to research. In fact, the enrichment of specific cell types is beneficial for enteroid-based research to increase the signal originating from a specific population of cells and to provide a larger population of cells of interest for downstream single cell analysis of enteroids, which is particularly beneficial when studying rare populations such as Paneth cells.…”

Utility Evaluation of Porcine Enteroids as PDCoV Infection Model in vitro

“…Culture of the MPIEs was carried out as recommended Thorne et al, 2018;van der Hee et al, 2018). Highly confluent enteroids were obtained after 7 days culture and dissociated into a single-cell suspension with EDTA-trypsin (Invitrogen, United States).…”

Transit-amplifying cells coordinate changes in intestinal epithelial cell-type composition

“…To understand how combinations of microenvironmental signals are interpreted to regulate tissue cell-type composition, we made use of a recently developed enteroid monolayer culture system. These cultures recapitulate key features of the intestinal epithelium including representation of each major intestinal epithelial cell-type (Lgr5 + stem, transit-amplifying, and differentiated secretory and absorptive cells), apicalbasolateral polarization, and self-organization 4 . Importantly, enteroid monolayers can be systematically perturbed and assayed via imaging.…”

“…Enteroid monolayers were derived as previously described 4 . Briefly, jejunum was isolated from male mice between 6-12 weeks of age.…”

“…Here, we address these questions in the context of the intestinal epithelium, a model renewing tissue 1,2,3 . Our approach builds on a previously established enteroid monolayer culture system that recapitulates key properties of the intestinal epithelium and is amenable to quantitative microscopy due to its 2D nature 4 . We combine enteroid culture with high-throughput combinatorial perturbation and quantitative imaging of stem, TA, and differentiated cell-types.…”