Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids

Dogs are increasingly recognized as valuable large animal models for understanding human intestinal diseases, as they naturally develop conditions similar to those in humans, such as Enterohemorrhagic E. coli, Clostridium difficile infection, inflammatory bowel disease, and ulcerative colitis. Given...

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Main Authors: Shino Yoshida, Meg Nakazawa, Minae Kawasaki, Yoko M. Ambrosini
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-12-01
Series:Frontiers in Veterinary Science
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fvets.2024.1483421/full
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author Shino Yoshida
Meg Nakazawa
Minae Kawasaki
Yoko M. Ambrosini
author_facet Shino Yoshida
Meg Nakazawa
Minae Kawasaki
Yoko M. Ambrosini
author_sort Shino Yoshida
collection DOAJ
description Dogs are increasingly recognized as valuable large animal models for understanding human intestinal diseases, as they naturally develop conditions similar to those in humans, such as Enterohemorrhagic E. coli, Clostridium difficile infection, inflammatory bowel disease, and ulcerative colitis. Given the similarity in gut flora between dogs and humans, canine in vitro intestinal models are ideal for translational research. However, conventional extracellular matrix-embedded organoids present challenges in accessing the lumen, which is critical for gut function. This study aimed to investigate the feasibility of inducing polarity reversal and differentiation in canine apical-out colonic organoids (colonoids), evaluate their barrier integrity, and visualize host-pathogen interactions. Our results demonstrated successful polarity reversal and differentiation induction while maintaining barrier integrity. Polarity reversal allowed for enhanced observation of host-pathogen interactions, facilitating visual assessments and membrane integrity evaluations using both pathogenic and nonpathogenic E. coli. This process led to the downregulation of stem cell marker LGR5 and upregulation of intestinal epithelial cell marker ALPI, indicating differentiation. Further differentiation was observed with the use of a differentiation culture medium, resulting in significant upregulation of ALPI and goblet cell marker MUC2. The findings suggest that apical-out canine colonoids can serve as physiologic and valuable models for studying the pathogenic mechanisms and clinical significance of intestinal diseases in dogs. This model has the potential to advance both canine and human gastrointestinal research, enhancing our understanding of gastrointestinal physiology and pathology and aiding in the development of novel therapeutics.
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publishDate 2024-12-01
publisher Frontiers Media S.A.
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spelling doaj-art-7f128acb31eb4dcb9435da4c7e81d51a2024-12-18T06:43:58ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692024-12-011110.3389/fvets.2024.14834211483421Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoidsShino YoshidaMeg NakazawaMinae KawasakiYoko M. AmbrosiniDogs are increasingly recognized as valuable large animal models for understanding human intestinal diseases, as they naturally develop conditions similar to those in humans, such as Enterohemorrhagic E. coli, Clostridium difficile infection, inflammatory bowel disease, and ulcerative colitis. Given the similarity in gut flora between dogs and humans, canine in vitro intestinal models are ideal for translational research. However, conventional extracellular matrix-embedded organoids present challenges in accessing the lumen, which is critical for gut function. This study aimed to investigate the feasibility of inducing polarity reversal and differentiation in canine apical-out colonic organoids (colonoids), evaluate their barrier integrity, and visualize host-pathogen interactions. Our results demonstrated successful polarity reversal and differentiation induction while maintaining barrier integrity. Polarity reversal allowed for enhanced observation of host-pathogen interactions, facilitating visual assessments and membrane integrity evaluations using both pathogenic and nonpathogenic E. coli. This process led to the downregulation of stem cell marker LGR5 and upregulation of intestinal epithelial cell marker ALPI, indicating differentiation. Further differentiation was observed with the use of a differentiation culture medium, resulting in significant upregulation of ALPI and goblet cell marker MUC2. The findings suggest that apical-out canine colonoids can serve as physiologic and valuable models for studying the pathogenic mechanisms and clinical significance of intestinal diseases in dogs. This model has the potential to advance both canine and human gastrointestinal research, enhancing our understanding of gastrointestinal physiology and pathology and aiding in the development of novel therapeutics.https://www.frontiersin.org/articles/10.3389/fvets.2024.1483421/fullorganoidcaninepolarity reversalapical-outintestinedifferentiation
spellingShingle Shino Yoshida
Meg Nakazawa
Minae Kawasaki
Yoko M. Ambrosini
Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids
Frontiers in Veterinary Science
organoid
canine
polarity reversal
apical-out
intestine
differentiation
title Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids
title_full Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids
title_fullStr Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids
title_full_unstemmed Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids
title_short Bacterial attachment and junctional transport function in induced apical-out polarized and differentiated canine intestinal organoids
title_sort bacterial attachment and junctional transport function in induced apical out polarized and differentiated canine intestinal organoids
topic organoid
canine
polarity reversal
apical-out
intestine
differentiation
url https://www.frontiersin.org/articles/10.3389/fvets.2024.1483421/full
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AT megnakazawa bacterialattachmentandjunctionaltransportfunctionininducedapicaloutpolarizedanddifferentiatedcanineintestinalorganoids
AT minaekawasaki bacterialattachmentandjunctionaltransportfunctionininducedapicaloutpolarizedanddifferentiatedcanineintestinalorganoids
AT yokomambrosini bacterialattachmentandjunctionaltransportfunctionininducedapicaloutpolarizedanddifferentiatedcanineintestinalorganoids