A18 BUILDING BETTER ENTEROIDS: A NOVEL STRATEGY FOR ENRICHING SECRETORY EPITHELIAL CELL SUBTYPES
Notice bibliographique
Résumé
Abstract Background Inflammatory bowel diseases (IBD) are chronic gastrointestinal disorders that affect more than 270,000 Canadians, including over 7,000 children. Inflammation arising from IBD severely damages intestinal epithelial cells (IECs), however their role in disease pathogenesis is not fully understood, in part due to a lack of in vitro systems that recapitulate the epithelium’s complex cellular composition. Enteroids are an in vitro model where primary IECs are isolated and cultured as 3D ‘mini guts.’ They offer distinct advantages over cell lines, however current protocols generate enteroids primarily composed of enterocytes that seldom contain rarer IEC subtypes (goblet, enteroendocrine, tuft and Paneth cells). These cells are responsible for the gut’s mucus, antimicrobial and hormone secretory functions, yet their role in the pathophysiology of IBD is unclear. Aims Manipulate cell differentiation pathways in mouse and human enteroids to increase the prevalence of secretory IECs, as well as determine if enteroids derived from pediatric IBD patients exhibit impaired responses to differentiation treatments. Methods Mouse enteroids were derived from ileal, cecal and colonic crypts of C57BL/6 mice while human enteroids were isolated from healthy or pediatric IBD patient intestinal biopsies. Enteroids were initially supplemented with Wnt signaling activators and the extracellular matrix modified to enhance enteroid culture “stemness”. Differentiation was induced by growth media modulation of the Notch and Wnt signaling pathways. Enteroids were analyzed via flow cytometry to quantify expression of secretory cell markers, while immunofluorescent and Peroidic acid Schiff/Alcian Blue staining was used to visualize goblet cells. Results “Stem” treatment potentiated Wnt signaling and enhanced enteroid “stemness” as measured by Lgr5 and CD44 expression. Comparatively, differentiation of these “stem” enteroids led to a larger relative increase in secretory markers. Differentiation treatment increased expression of goblet cell markers (Muc2 and lectin) in the “stem” treated mouse cecal and colonic enteroids, but not in ileal enteroids. Notch inhibition produced increased expression of lysozyme, a Paneth cell marker, in all enteroids. A similar increase in secretory cell numbers was observed in control human enteroids following differentiation treatment. In contrast, enteroids derived from pediatric IBD patients displayed irregular differentation responses to treatment. Conclusions We demonstrate that manipulation of cell differentiation pathways increases the number of secretory IEC subtypes within enteroids. Furthermore, these pro-secretory cell responses differ in IBD patient enteroids, indicating that an alteration of the targeted cell signaling pathways may be linked to IBD pathogenesis. Funding Agencies CCC, CIHRBCCHRI Summer Studentship
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».