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Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis

BACKGROUND & AIMS: Epithelial disruption in eosinophilic esophagitis (EoE) encompasses both impaired differentiation and diminished barrier integrity. We have shown that lysyl oxidase (LOX), a collagen cross-linking enzyme, is upregulated in the esophageal epithelium in EoE. However, the functio...

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Autores principales: Sasaki, Masaru, Hara, Takeo, Wang, Joshua X., Zhou, Yusen, Kennedy, Kanak V., Umeweni, Nicole N., Alston, Maiya A., Spergel, Zachary C., Nakagawa, Ritsu, Mcmillan, Emily A., Whelan, Kelly A., Karakasheva, Tatiana A., Hamilton, Kathryn E., Ruffner, Melanie A., Muir, Amanda B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10081173/
https://www.ncbi.nlm.nih.gov/pubmed/37034590
http://dx.doi.org/10.1101/2023.03.27.534387
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author Sasaki, Masaru
Hara, Takeo
Wang, Joshua X.
Zhou, Yusen
Kennedy, Kanak V.
Umeweni, Nicole N.
Alston, Maiya A.
Spergel, Zachary C.
Nakagawa, Ritsu
Mcmillan, Emily A.
Whelan, Kelly A.
Karakasheva, Tatiana A.
Hamilton, Kathryn E.
Ruffner, Melanie A.
Muir, Amanda B.
author_facet Sasaki, Masaru
Hara, Takeo
Wang, Joshua X.
Zhou, Yusen
Kennedy, Kanak V.
Umeweni, Nicole N.
Alston, Maiya A.
Spergel, Zachary C.
Nakagawa, Ritsu
Mcmillan, Emily A.
Whelan, Kelly A.
Karakasheva, Tatiana A.
Hamilton, Kathryn E.
Ruffner, Melanie A.
Muir, Amanda B.
author_sort Sasaki, Masaru
collection PubMed
description BACKGROUND & AIMS: Epithelial disruption in eosinophilic esophagitis (EoE) encompasses both impaired differentiation and diminished barrier integrity. We have shown that lysyl oxidase (LOX), a collagen cross-linking enzyme, is upregulated in the esophageal epithelium in EoE. However, the functional roles of LOX in the esophageal epithelium remains unknown. METHODS: We investigated roles for LOX in the human esophageal epithelium using 3-dimensional organoid and air-liquid interface cultures stimulated with interleukin (IL)-13 to recapitulate the EoE inflammatory milieu, followed by single-cell RNA sequencing, quantitative reverse transcription-polymerase chain reaction, western blot, histology, and functional analyses of barrier integrity. RESULTS: Single-cell RNA sequencing analysis on patient-derived organoids revealed that LOX was induced by IL-13 in differentiated cells. LOX-overexpressing organoids demonstrated suppressed basal and upregulated differentiation markers. Additionally, LOX overexpression enhanced junctional protein genes and transepithelial electrical resistance. LOX overexpression restored the impaired differentiation and barrier function, including in the setting of IL-13 stimulation. Transcriptome analyses on LOX-overexpressing organoids identified enriched bone morphogenetic protein (BMP) signaling pathway compared to wild type organoids. Particularly, LOX overexpression increased BMP2 and decreased BMP antagonist follistatin. Finally, we found that BMP2 treatment restored the balance of basal and differentiated cells. CONCLUSIONS: Our data support a model whereby LOX exhibits non-canonical roles as a signaling molecule important for epithelial homeostasis in the setting of inflammation via activation of BMP pathway in esophagus. The LOX/BMP axis may be integral in esophageal epithelial differentiation and a promising target for future therapies.
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spelling pubmed-100811732023-04-08 Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis Sasaki, Masaru Hara, Takeo Wang, Joshua X. Zhou, Yusen Kennedy, Kanak V. Umeweni, Nicole N. Alston, Maiya A. Spergel, Zachary C. Nakagawa, Ritsu Mcmillan, Emily A. Whelan, Kelly A. Karakasheva, Tatiana A. Hamilton, Kathryn E. Ruffner, Melanie A. Muir, Amanda B. bioRxiv Article BACKGROUND & AIMS: Epithelial disruption in eosinophilic esophagitis (EoE) encompasses both impaired differentiation and diminished barrier integrity. We have shown that lysyl oxidase (LOX), a collagen cross-linking enzyme, is upregulated in the esophageal epithelium in EoE. However, the functional roles of LOX in the esophageal epithelium remains unknown. METHODS: We investigated roles for LOX in the human esophageal epithelium using 3-dimensional organoid and air-liquid interface cultures stimulated with interleukin (IL)-13 to recapitulate the EoE inflammatory milieu, followed by single-cell RNA sequencing, quantitative reverse transcription-polymerase chain reaction, western blot, histology, and functional analyses of barrier integrity. RESULTS: Single-cell RNA sequencing analysis on patient-derived organoids revealed that LOX was induced by IL-13 in differentiated cells. LOX-overexpressing organoids demonstrated suppressed basal and upregulated differentiation markers. Additionally, LOX overexpression enhanced junctional protein genes and transepithelial electrical resistance. LOX overexpression restored the impaired differentiation and barrier function, including in the setting of IL-13 stimulation. Transcriptome analyses on LOX-overexpressing organoids identified enriched bone morphogenetic protein (BMP) signaling pathway compared to wild type organoids. Particularly, LOX overexpression increased BMP2 and decreased BMP antagonist follistatin. Finally, we found that BMP2 treatment restored the balance of basal and differentiated cells. CONCLUSIONS: Our data support a model whereby LOX exhibits non-canonical roles as a signaling molecule important for epithelial homeostasis in the setting of inflammation via activation of BMP pathway in esophagus. The LOX/BMP axis may be integral in esophageal epithelial differentiation and a promising target for future therapies. Cold Spring Harbor Laboratory 2023-03-28 /pmc/articles/PMC10081173/ /pubmed/37034590 http://dx.doi.org/10.1101/2023.03.27.534387 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Sasaki, Masaru
Hara, Takeo
Wang, Joshua X.
Zhou, Yusen
Kennedy, Kanak V.
Umeweni, Nicole N.
Alston, Maiya A.
Spergel, Zachary C.
Nakagawa, Ritsu
Mcmillan, Emily A.
Whelan, Kelly A.
Karakasheva, Tatiana A.
Hamilton, Kathryn E.
Ruffner, Melanie A.
Muir, Amanda B.
Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis
title Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis
title_full Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis
title_fullStr Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis
title_full_unstemmed Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis
title_short Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis
title_sort lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10081173/
https://www.ncbi.nlm.nih.gov/pubmed/37034590
http://dx.doi.org/10.1101/2023.03.27.534387
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