Differential Modulation by Akkermansia muciniphila and Faecalibacterium prausnitzii of Host Peripheral Lipid Metabolism and Histone Acetylation in Mouse Gut Organoids

Lukovac, Sabina; Belzer, Clara; Pellis, Linette; Keijser, Bart J. F.; Vos, Willem M. de; Montijn, R.C.; Roeselers, Guus

doi:10.1128/mbio.01438-14

Cited by 382 publications

(251 citation statements)

References 40 publications

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“…Recently, others have utilized mouse-derived enteroids (epithelium-only organoids) to study host-microbe interactions (29,30). A key difference between those reports and the data presented here is that our system utilizes organoids derived from human cells rather than mouse intestinal crypts.…”

Section: Discussioncontrasting

confidence: 44%

Persistence and Toxin Production by Clostridium difficile within Human Intestinal Organoids Result in Disruption of Epithelial Paracellular Barrier Function

et al. 2015

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f Clostridium difficile is the leading cause of infectious nosocomial diarrhea. The pathogenesis of C. difficile infection (CDI) results from the interactions between the pathogen, intestinal epithelium, host immune system, and gastrointestinal microbiota. Previous studies of the host-pathogen interaction in CDI have utilized either simple cell monolayers or in vivo models. While much has been learned by utilizing these approaches, little is known about the direct interaction of the bacterium with a complex host epithelium. Here, we asked if human intestinal organoids (HIOs), which are derived from pluripotent stem cells and demonstrate small intestinal morphology and physiology, could be used to study the pathogenesis of the obligate anaerobe C. difficile. Vegetative C. difficile, microinjected into the lumen of HIOs, persisted in a viable state for up to 12 h. Upon colonization with C. difficile VPI 10463, the HIO epithelium is markedly disrupted, resulting in the loss of paracellular barrier function. Since similar effects were not observed when HIOs were colonized with the nontoxigenic C. difficile strain F200, we directly tested the role of toxin using TcdA and TcdB purified from VPI 10463. We show that the injection of TcdA replicates the disruption of the epithelial barrier function and structure observed in HIOs colonized with viable C. difficile. C lostridium difficile is an anaerobic, spore-forming bacterium that is the leading cause of infectious nosocomial diarrhea and is responsible for over 14,000 deaths annually (1). Human exposure to C. difficile results in a range of manifestations, from asymptomatic colonization, to diarrhea, to lethal toxic megacolon. Various models have been used to study C. difficile infection (CDI), including in vitro models using transformed cell lines and a variety of in vivo models (2-5). In vitro cell culture models are limited in their ability to recapitulate complexities of the human gastrointestinal tract, and detailed, real-time study of the mucosal epithelium during infection in an animal model is technically challenging.Human intestinal organoids (HIOs) are three-dimensional spheroids of human epithelium generated through directed differentiation of human pluripotent stem cells (hPSCs), which include human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). HIOs contain both mesenchymal and epithelial tissues that are structurally arranged around a central luminal cavity. The epithelial compartment of the HIO possesses an array of small intestinal cell types, including absorptive enterocytes and secretory Paneth, goblet, and enteroendocrine cells, in addition to Lgr5 ϩ intestinal stem cells (6). HIOs have been used to model features of embryonic development, viral infection, and inflammatory bowel disease (7-9). Due to their similarity to the human gastrointestinal tract, HIOs serve as a tractable and physiologically relevant model of the human intestine.We sought to use HIOs to study the interaction between C. difficile and complex human epit...

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Section: Discussioncontrasting

confidence: 44%

Persistence and Toxin Production by Clostridium difficile within Human Intestinal Organoids Result in Disruption of Epithelial Paracellular Barrier Function

et al. 2015

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“…Enteroendocrine function such as incretin release will be valuable for studying microbiome product-intestinal interactions [35]. Overall, these studies confirm that generating HIO is a viable option for producing intestinal epithelial-like cells from human iPSCs.…”

Section: Discussionsupporting

confidence: 60%

Directed differentiation protocols for successful human intestinal organoids derived from multiple induced pluripotent stem cell lines

Kauffman¹,

Ekert²,

Gyurdieva³

et al. 2015

Stem Cell Biol Res

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Background: Relatively little has been reported comparing the ability of different induced pluripotent stem cells (iPSCs) and protocols to derive human intestinal organoids (HIO), although there is potential to supply HIO for translational research and regenerative medicine. In view of the time and effort required to differentiate HIO, protocols for differentiation were compared and five iPSC lines, produced by retroviral or non-viral reprogramming methods, were concurrently differentiated to HIO to evaluate the robustness and repeatability of using cellular markers by flow cytometry and immunohistochemistry. Methods: iPSCs were differentiated to definitive endoderm by Activin A treatment (Protocol I) or Myostatin (GDF8), GSK-3β inhibitor (CHIR99021) and B27 supplement (Protocol II) and then differentiated to hindgut using FGF4 and WNT3A (Protocol I), or KGF and Retinoic Acid (Protocol II) resulting in spheroids that were differentiated to HIO in 3-dimensional culture in matrigel containing EGF, Noggin, and R-Spondin1. Results: Definitive endoderm markers (CXCR4 and SOX17) were similar using both protocols and in all cell lines, but in their absence, spheroids to not develop. HIO derived from excised hindgut spheroids (Protocol II) showed more consistent expression of epithelial markers E-Cadherin, CDX2, villin, and chromogranin A. Morphological characteristics such as budding are predictive of robust enterocyte differentiation (villin positive) in HIO. Conclusions: Substituting different members of the growth factor families (e.g., TGFβ) is equally or more effective for producing epithelial lineages in HIO. Although there are no early quantitative predictors of level of success, all reprogrammed somatic cells could be differentiated to produce HIO. Several morphological characteristics (excisable spheroids and budding in HIO) were associated with HIO success. This could improve cost-effectiveness and ease of differentiation as HIO become more commonly available as translational 3D models of the gut.

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“…Bacterial metabolites like short-chain fatty acids (SCFA) produced by Akkermansia muciniphila and Faecalibacterium prausnitzii also influence immunity. For instance, butyrate from these organisms increases the differentiation of Tregs in the colon (26,27).…”

Section: Microbial and Dietary Influences On Intestinal Immunitymentioning

confidence: 99%

Immunologic impact of the intestine in metabolic disease

Winer¹,

Winer²,

Dranse³

et al. 2017

Journal of Clinical Investigation

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Differential Modulation by Akkermansia muciniphila and Faecalibacterium prausnitzii of Host Peripheral Lipid Metabolism and Histone Acetylation in Mouse Gut Organoids

Cited by 382 publications

References 40 publications

Persistence and Toxin Production by Clostridium difficile within Human Intestinal Organoids Result in Disruption of Epithelial Paracellular Barrier Function

Persistence and Toxin Production by Clostridium difficile within Human Intestinal Organoids Result in Disruption of Epithelial Paracellular Barrier Function

Directed differentiation protocols for successful human intestinal organoids derived from multiple induced pluripotent stem cell lines

Immunologic impact of the intestine in metabolic disease

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