Autologous organoid co-culture model reveals T cell-driven epithelial cell death in Crohn’s Disease

Hammoudi, Nassim; Hamoudi, Sarah; Bonnereau, Julie; Bottois, Hugo; Pérez, Kevin; Bezault, Madeleine; Hassid, Déborah; Chardiny, Victor; Grand, Céline; Gergaud, Brice; Bonnet, Joëlle; Chedouba, Leïla; Minh, My-Linh Tran; Gornet, Jean–Marc; Baudry, Clotilde; Maggiori, Léon; Toubert, Antoine; McBride, Jacqueline; Brochier, Camille; Neighbors, Margaret; Bourhis, Lionel Le; Allez, Matthieu

doi:10.3389/fimmu.2022.1008456

Cited by 14 publications

(13 citation statements)

References 48 publications

(50 reference statements)

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“…Our findings demonstrate that E-Cadherin co-stimulates CD8 + T cells as evident by the upregulation of CD69 and the secretion of cytotoxic mediators and suggest that this mechanism might be important in the context of IBD. Together with a previous report showing that the interaction of αEβ7 with E-Cadherin also drives T cell infiltration to epithelial organoids 36 , it emerges that αEβ7 is a key modulator of epithelial-lymphocyte crosstalk. However, while etrolizumab seems to be able to block adhesive lympho-epithelial interaction and cytotoxicity resulting from direct cell-to-cell contact of T lymphocytes with the epithelium 36 , our data suggest that co-stimulatory pathways leading to the secretion of soluble cytotoxic mediators are not sufficiently blocked and we therefore hypothesize that these mediators might cause epithelial cell death in the inflamed gut in a paracrine manner as demonstrated in the literature 37 , 38 .…”

Section: Discussionsupporting

confidence: 62%

Etrolizumab-s fails to control E-Cadherin-dependent co-stimulation of highly activated cytotoxic T cells

Wiendl,

Dedden,

Liu

et al. 2024

Nat Commun

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Despite promising preclinical and earlier clinical data, a recent phase III trial on the anti-β7 integrin antibody etrolizumab in Crohn’s disease (CD) did not reach its primary endpoint. The mechanisms leading to this outcome are not well understood. Here we characterize the β7+ T cell compartment from patients with CD in comparison to cells from individuals without inflammatory bowel disease. By flow cytometric, transcriptomic and functional profiling of circulating T cells, we find that triple-integrin-expressing (α4+β7+β1hi) T cells have the potential to home to the gut despite α4β7 blockade and have a specific cytotoxic signature. A subset of triple-integrin-expressing cells readily acquires αE expression and could be co-stimulated via E-Cadherin-αEβ7 interactions in vitro. Etrolizumab-s fails to block such αEβ7 signalling at high levels of T cell stimulation. Consistently, in CD patients treated with etrolizumab, T cell activation correlates with cytotoxic signatures. Collectively, our findings might add one important piece to the puzzle to explain phase III trial results with etrolizumab, while they also highlight that αEβ7 remains an interesting target for future therapeutic approaches in inflammatory bowel disease.

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Section: Discussionsupporting

confidence: 62%

Etrolizumab-s fails to control E-Cadherin-dependent co-stimulation of highly activated cytotoxic T cells

Wiendl,

Dedden,

Liu

et al. 2024

Nat Commun

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“…For instance, organoid‐immune cell co‐cultures (Support Protocol 3) could shed light on the interplay between MP/NP uptake by intestinal epithelial cells and immune responses by leukocytes (Rubio et al., 2020; Staab et al., 2020). Models of organoid‐immune cell co‐cultures have been established and used in investigation of inflammatory responses in the intestinal system (Hammoudi et al., 2022; Schreurs et al., 2021). Specifically, the presence of MP/NP exacerbated the severity of inflammatory bowel disease (IBD) and Crohn's Disease (CD) (Yan et al., 2022; Zhao et al., 2023).…”

Section: Commentarymentioning

confidence: 99%

Impact of Micro‐ and Nano‐Plastics on Human Intestinal Organoid‐Derived Epithelium

Wang,

Iglesias‐Ledon,

Bishop

et al. 2024

Current Protocols

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The development of patient‐derived intestinal organoids represents an invaluable model for simulating the native human intestinal epithelium. These stem cell‐rich cultures outperform commonly used cell lines like Caco‐2 and HT29‐MTX in reflecting the cellular diversity of the native intestinal epithelium after differentiation. In our recent study examining the effects of polystyrene (PS), microplastics (MPs), and nanoplastics (NPs), widespread pollutants in our environment and food chain, on the human intestinal epithelium, these organoids have been instrumental in elucidating the absorption mechanisms and potential biological impacts of plastic particles. Building on previously established protocols in human intestinal organoid culture, we herein detail a streamlined protocol for the cultivation, differentiation, and generation of organoid‐derived monolayers. This protocol is tailored to generate monolayers incorporating microfold cells (M cells), key for intestinal particle uptake but often absent in current in vitro models. We provide validated protocols for the characterization of MPs/NPs via scanning electron microscopy (SEM) for detailed imaging and their introduction to intestinal epithelial monolayer cells via confocal immunostaining. Additionally, protocols to test the impacts of MP/NP exposure on the functions of the intestinal barrier using transendothelial electrical resistance (TEER) measurements and assessing inflammatory responses using cytokine profiling are detailed. Overall, our protocols enable the generation of human intestinal organoid monolayers, complete with the option of including or excluding M cells, offering crucial techniques for observing particle uptake and identifying inflammatory responses in intestinal epithelial cells to advance our knowledge of the potential effects of plastic pollution on human gut health. These approaches are also amendable to the study of other gut‐related chemical and biological exposures and physiological responses due to the robust nature of the systems. © 2024 Wiley Periodicals LLC.Basic Protocol 1: Human intestinal organoid culture and generation of monolayers with and without M cellsSupport Protocol 1: Culture of L‐WRN and production of WRN‐conditioned mediumSupport Protocol 2: Neuronal cell culture and integration into intestinal epitheliumSupport Protocol 3: Immune cell culture and integration into intestinal epitheliumBasic Protocol 2: Scanning electron microscopy: sample preparation and imagingBasic Protocol 3: Immunostaining and confocal imaging of MP/NP uptake in organoid‐derived monolayersBasic Protocol 4: Assessment of intestinal barrier function via TEER measurementsBasic Protocol 5: Cytokine profiling using ELISA post‐MP/NP exposure.

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“…T cells are known to drive the pathogenesis of intestinal inflammatory disorders due to excessive cytotoxicity and production of pro-inflammatory cytokines. T cells and organoids have been co-cultured indirectly using transwell systems (with T cells cultured in the basolateral chamber) [87], or directly by embedding both populations in an ECM [88,89]. Using the indirect coculture method, it was demonstrated that secretion of inflammatory mediators by T cells is sufficient to destroy colonoid monolayers [87].…”

Section: Epithelial Cell-cell Interactionsmentioning

confidence: 99%

Human intestinal organoids: Modeling gastrointestinal physiology and immunopathology — current applications and limitations

Flood,

Hanrahan,

Nally

et al. 2023

Eur J Immunol

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Human intestinal organoids are an ideal model system to study gastrointestinal physiology and immunopathology. Altered physiology and mucosal immune response are hallmarks of numerous intestinal functional and inflammatory diseases, including inflammatory bowel disease (IBD), coeliac disease, irritable bowel syndrome (IBS), and obesity. These conditions impact the normal epithelial functions of the intestine, such as absorption, barrier function, secretion, and host‐microbiome communication. They are accompanied by characteristic intestinal symptoms and have significant societal, economic, and healthcare burdens. To develop new treatment options, cutting‐edge research is required to investigate their aetiology and pathology. Human intestinal organoids derived from patient tissue recapitulate the key physiological and immunopathological aspects of these conditions, providing a promising platform for elucidating disease mechanisms. This review will summarise recent reports on patient derived human small intestinal and colonic organoids and highlight how these models have been used to study intestinal epithelial functions in the context of inflammation, altered physiology and immune response. Furthermore, it will elaborate on the various organoid systems in use and the techniques/assays currently available to study epithelial functions. Finally, it will conclude by discussing the limitations and future perspectives of organoid technology.This article is protected by copyright. All rights reserved

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Autologous organoid co-culture model reveals T cell-driven epithelial cell death in Crohn’s Disease

Cited by 14 publications

References 48 publications

Etrolizumab-s fails to control E-Cadherin-dependent co-stimulation of highly activated cytotoxic T cells

Etrolizumab-s fails to control E-Cadherin-dependent co-stimulation of highly activated cytotoxic T cells

Impact of Micro‐ and Nano‐Plastics on Human Intestinal Organoid‐Derived Epithelium

Human intestinal organoids: Modeling gastrointestinal physiology and immunopathology — current applications and limitations

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