Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis

Sasaki, M.; Hara, T.; Wang, J. X.; Zhou, Y.; Kennedy, K. V.; Umeweni, N. N.; Alston, M. A.; Spergel, Z. C.; Nakagawa, R.; McMillan, E. A.; Whelan, K. A.; Karakasheva, T. A.; Hamilton, K. E.; Ruffner, M. A.; Muir, A. B.

doi:10.1101/2023.03.27.534387

bioRxiv · 10.1101/2023.03.27.534387

Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis

Sasaki, M.·Hara, T.·Wang, J. X.·Zhou, Y.·Kennedy, K. V.·Umeweni, N. N.·Alston, M. A.·Spergel, Z. C.·Nakagawa, R.·McMillan, E. A.·Whelan, K. A.·Karakasheva, T. A.·Hamilton, K. E.·Ruffner, M. A.·Muir, A. B.

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Abstract

Background & AimsEpithelial 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. MethodsWe 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. ResultsSingle-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. ConclusionsOur 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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Sasaki, M., Hara, T., Wang, J. X., Zhou, Y., Kennedy, K. V., Umeweni, N. N., Alston, M. A., Spergel, Z. C., Nakagawa, R., McMillan, E. A., Whelan, K. A., Karakasheva, T. A., Hamilton, K. E., Ruffner, M. A., Muir, A. B.. 2023-03-28. Lysyl oxidase regulates epithelial differentiation and barrier integrity in eosinophilic esophagitis. https://doi.org/10.1101/2023.03.27.534387

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