Dataset Information

Chemically Induced Paneth Cells Provide Superior In Vivo Mimic to Organoid-derived Analog

ABSTRACT: Paneth cells (PCs) are post-mitotic secretory cells of the small intestine that produce antimicrobials, growth factors, and cytokines. Multiple diseases have been linked to alterations in PC function. Past studies of PCs have relied on in vivo models, and limited in vitro models, including organoids. While useful, whether the in vitro models recapitulate in vivo biology is an important, yet unanswered question. Here, we present a bona fide in vitro PC-enriched population derived via chemically-induced (CI) differentiation of primary murine adult stem cells. We define the identity of PCs in our system and compare to PCs of established organoid systems and those found in vivo. Importantly, we demonstrate stimulant-induced and microbe-responsive antimicrobial function of our CI-PCs – hallmark functions of PCs in vivo. Thus, we define and produce the most physiologically-relevant in vitro PC population to date, providing a superior tool to study cellular development, host-microbe interactions, and inflammatory disease.

ORGANISM(S): Mus musculus

PROVIDER: GSE100274 | GEO | 2018/04/02

REPOSITORIES: GEO

ACCESS DATA

Similar Datasets

Project description:Mead BE, Ordovas-Montanes J, Braun AP, Levy LE, Bhargava P, Szucs MJ, Ammendolia DA, MacMullan MA, Zheng Y, Yin X, Hughes TK, Wadsworth MH, Ahmad R, Rakoff-Nahoum S, Carr SA, Langer R, Collins JJ, Shalek AK, Karp JM. BMC Biology 2018. Background: Single-cell genomic methods now provide unprecedented resolution for characterizing the component cell types and states of tissues, such as the epithelial subsets of the gastrointestinal tract. Nevertheless, functional studies of these subsets at scale require faithful in vitro models of identified in vivo biology. While intestinal organoids have been invaluable in providing mechanistic insights in vitro, the extent to which organoid-derived cell types recapitulate their in vivo counterparts remains formally untested, with no systematic approach for improving model fidelity. Results: Here, we present a generally applicable framework that utilizes massively-parallel single-cell RNA-seq to compare cell types and states found in vivo to those of in vitro models, such as organoids. Furthermore, we leverage identified discrepancies to improve model fidelity. Using the Paneth cell (PC), which supports the stem cell niche and produces the largest diversity of antimicrobials in the small intestine, as an exemplar, we uncover fundamental gene expression differences in lineage-defining genes between in vivo PCs and those of the current in vitro organoid model. With this information, we nominate a molecular intervention to rationally improve the physiological fidelity of our in vitro PCs. We then perform transcriptomic, cytometric, morphologic, and proteomic characterization, and demonstrate functional (antimicrobial activity, niche support) improvements in Paneth cell physiology. Conclusions: Our systematic approach provides a simple workflow for identifying the limitations of in vitro models and enhancing their physiological fidelity. Using adult stem cell-derived PCs within intestinal organoids as a model system, we successfully benchmark organoid representation relative to in vivo of a specialized cell type and use this comparison to generate a functionally improved in vitro PC population. We predict that the generation of rationally-improved cellular models will facilitate mechanistic exploration of specific disease-associated genes in their respective cell types.

Dataset Information

Chemically Induced Paneth Cells Provide Superior In Vivo Mimic to Organoid-derived Analog

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure