Development of New Models of Oral Mucosa to Investigate the Impact of the Structure of Transmembrane Mucin-1 on the Mucosal Pellicle Formation and Its Physicochemical Properties

Clément Nivet; Irma Custovic; Laure Avoscan; Floris J. Bikker; Aline Bonnotte; Eric Bourillot; Loïc Briand; Hélène Brignot; Jean Marie Heydel; Noémie Herrmann; Mélanie Lelièvre; Eric Lesniewska; Fabrice Neiers; Olivier Piétrement; Mathieu Schwartz; Christine Belloir; Francis Canon

doi:10.3390/biomedicines12010139

Development of New Models of Oral Mucosa to Investigate the Impact of the Structure of Transmembrane Mucin-1 on the Mucosal Pellicle Formation and Its Physicochemical Properties

Clément Nivet, Irma Custovic, Laure Avoscan, Floris J. Bikker, Aline Bonnotte, Eric Bourillot, Loïc Briand, Hélène Brignot, Jean Marie Heydel, Noémie Herrmann, Mélanie Lelièvre, Eric Lesniewska, Fabrice Neiers, Olivier Piétrement, Mathieu Schwartz, Christine Belloir, Francis Canon^*

^*Corresponding author for this work

Oral Biochemistry

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

The mucosal pellicle (MP) is a biological film protecting the oral mucosa. It is composed of bounded salivary proteins and transmembrane mucin MUC1 expressed by oral epithelial cells. Previous research indicates that MUC1 expression enhances the binding of the main salivary protein forming the MP, MUC5B. This study investigated the influence of MUC1 structure on MP formation. A TR146 cell line, which does not express MUC1 natively, was stably transfected with genes coding for three MUC1 isoforms differing in the structure of the two main extracellular domains: the VNTR domain, exhibiting a variable number of tandem repeats, and the SEA domain, maintaining the two bound subunits of MUC1. Semi-quantification of MUC1 using dot blot chemiluminescence showed comparable expression levels in all transfected cell lines. Semi-quantification of MUC5B by immunostaining after incubation with saliva revealed that MUC1 expression significantly increased MUC5B adsorption. Neither the VNTR domain nor the SEA domain was influenced MUC5B anchoring, suggesting the key role of the MUC1 N-terminal domain. AFM-IR nanospectroscopy revealed discernible shifts indicative of changes in the chemical properties at the cell surface due to the expression of the MUC1 isoform. Furthermore, the observed chemical shifts suggest the involvement of hydrophobic effects in the interaction between MUC1 and salivary proteins.

Original language	English
Article number	139
Pages (from-to)	1-21
Number of pages	21
Journal	Biomedicines
Volume	12
Issue number	1
Early online date	9 Jan 2024
DOIs	https://doi.org/10.3390/biomedicines12010139
Publication status	Published - Jan 2024

Bibliographical note

This article belongs to the Special Issue: Models for Oral Biology Research 2.0.

Funding Information:
This study was funded by the French National Research Agency (Grant MACARON ANR-20-CE21-002) and by Conseil Regional de Bourgogne Franche-Comté (France) (AAP Région 2020—ANER—Projet AFMdynDNA, AAP Région 2020 DNA-Heritage, AAP Région 2021—Projet MétaboFlav). This work has also been supported by the EIPHI Graduate School (contract ANR-17-EURE-0002). The French Ministry of Higher Education and Research provided the PhD fellowship for Clément Nivet.

Publisher Copyright:
© 2024 by the authors.

Funding

This study was funded by the French National Research Agency (Grant MACARON ANR-20-CE21-002) and by Conseil Regional de Bourgogne Franche-Comté (France) (AAP Région 2020—ANER—Projet AFMdynDNA, AAP Région 2020 DNA-Heritage, AAP Région 2021—Projet MétaboFlav). This work has also been supported by the EIPHI Graduate School (contract ANR-17-EURE-0002). The French Ministry of Higher Education and Research provided the PhD fellowship for Clément Nivet.

Funders	Funder number
ANER
EIPHI	ANR-17-EURE-0002
Association of Academic Physiatrists
Agence Nationale de la Recherche	MACARON ANR-20-CE21-002
Agence Nationale de la Recherche
Ministère de l'Education Nationale, de l'Enseignement Superieur et de la Recherche
Conseil régional de Bourgogne-Franche-Comté

Keywords

AFM-IR
MUC1
MUC5B
mucosal pellicle
oral mucosa
salivary protein interactions

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10.3390/biomedicines12010139

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Nivet, C., Custovic, I., Avoscan, L., Bikker, F. J., Bonnotte, A., Bourillot, E., Briand, L., Brignot, H., Heydel, J. M., Herrmann, N., Lelièvre, M., Lesniewska, E., Neiers, F., Piétrement, O., Schwartz, M., Belloir, C., & Canon, F. (2024). Development of New Models of Oral Mucosa to Investigate the Impact of the Structure of Transmembrane Mucin-1 on the Mucosal Pellicle Formation and Its Physicochemical Properties. Biomedicines, 12(1), 1-21. Article 139. https://doi.org/10.3390/biomedicines12010139

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title = "Development of New Models of Oral Mucosa to Investigate the Impact of the Structure of Transmembrane Mucin-1 on the Mucosal Pellicle Formation and Its Physicochemical Properties",

abstract = "The mucosal pellicle (MP) is a biological film protecting the oral mucosa. It is composed of bounded salivary proteins and transmembrane mucin MUC1 expressed by oral epithelial cells. Previous research indicates that MUC1 expression enhances the binding of the main salivary protein forming the MP, MUC5B. This study investigated the influence of MUC1 structure on MP formation. A TR146 cell line, which does not express MUC1 natively, was stably transfected with genes coding for three MUC1 isoforms differing in the structure of the two main extracellular domains: the VNTR domain, exhibiting a variable number of tandem repeats, and the SEA domain, maintaining the two bound subunits of MUC1. Semi-quantification of MUC1 using dot blot chemiluminescence showed comparable expression levels in all transfected cell lines. Semi-quantification of MUC5B by immunostaining after incubation with saliva revealed that MUC1 expression significantly increased MUC5B adsorption. Neither the VNTR domain nor the SEA domain was influenced MUC5B anchoring, suggesting the key role of the MUC1 N-terminal domain. AFM-IR nanospectroscopy revealed discernible shifts indicative of changes in the chemical properties at the cell surface due to the expression of the MUC1 isoform. Furthermore, the observed chemical shifts suggest the involvement of hydrophobic effects in the interaction between MUC1 and salivary proteins.",

keywords = "AFM-IR, MUC1, MUC5B, mucosal pellicle, oral mucosa, salivary protein interactions",

author = "Cl{\'e}ment Nivet and Irma Custovic and Laure Avoscan and Bikker, {Floris J.} and Aline Bonnotte and Eric Bourillot and Lo{\"i}c Briand and H{\'e}l{\`e}ne Brignot and Heydel, {Jean Marie} and No{\'e}mie Herrmann and M{\'e}lanie Leli{\`e}vre and Eric Lesniewska and Fabrice Neiers and Olivier Pi{\'e}trement and Mathieu Schwartz and Christine Belloir and Francis Canon",

note = "This article belongs to the Special Issue: Models for Oral Biology Research 2.0. Funding Information: This study was funded by the French National Research Agency (Grant MACARON ANR-20-CE21-002) and by Conseil Regional de Bourgogne Franche-Comt{\'e} (France) (AAP R{\'e}gion 2020—ANER—Projet AFMdynDNA, AAP R{\'e}gion 2020 DNA-Heritage, AAP R{\'e}gion 2021—Projet M{\'e}taboFlav). This work has also been supported by the EIPHI Graduate School (contract ANR-17-EURE-0002). The French Ministry of Higher Education and Research provided the PhD fellowship for Cl{\'e}ment Nivet. Publisher Copyright: {\textcopyright} 2024 by the authors.",

year = "2024",

month = jan,

doi = "10.3390/biomedicines12010139",

language = "English",

volume = "12",

pages = "1--21",

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Nivet, C, Custovic, I, Avoscan, L, Bikker, FJ, Bonnotte, A, Bourillot, E, Briand, L, Brignot, H, Heydel, JM, Herrmann, N, Lelièvre, M, Lesniewska, E, Neiers, F, Piétrement, O, Schwartz, M, Belloir, C & Canon, F 2024, 'Development of New Models of Oral Mucosa to Investigate the Impact of the Structure of Transmembrane Mucin-1 on the Mucosal Pellicle Formation and Its Physicochemical Properties', Biomedicines, vol. 12, no. 1, 139, pp. 1-21. https://doi.org/10.3390/biomedicines12010139

Development of New Models of Oral Mucosa to Investigate the Impact of the Structure of Transmembrane Mucin-1 on the Mucosal Pellicle Formation and Its Physicochemical Properties. / Nivet, Clément; Custovic, Irma; Avoscan, Laure et al.
In: Biomedicines, Vol. 12, No. 1, 139, 01.2024, p. 1-21.

Research output: Contribution to Journal › Article › Academic › peer-review

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AU - Nivet, Clément

AU - Custovic, Irma

AU - Avoscan, Laure

AU - Bikker, Floris J.

AU - Bonnotte, Aline

AU - Bourillot, Eric

AU - Briand, Loïc

AU - Brignot, Hélène

AU - Heydel, Jean Marie

AU - Herrmann, Noémie

AU - Lelièvre, Mélanie

AU - Lesniewska, Eric

AU - Neiers, Fabrice

AU - Piétrement, Olivier

AU - Schwartz, Mathieu

AU - Belloir, Christine

AU - Canon, Francis

N1 - This article belongs to the Special Issue: Models for Oral Biology Research 2.0. Funding Information: This study was funded by the French National Research Agency (Grant MACARON ANR-20-CE21-002) and by Conseil Regional de Bourgogne Franche-Comté (France) (AAP Région 2020—ANER—Projet AFMdynDNA, AAP Région 2020 DNA-Heritage, AAP Région 2021—Projet MétaboFlav). This work has also been supported by the EIPHI Graduate School (contract ANR-17-EURE-0002). The French Ministry of Higher Education and Research provided the PhD fellowship for Clément Nivet. Publisher Copyright: © 2024 by the authors.

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N2 - The mucosal pellicle (MP) is a biological film protecting the oral mucosa. It is composed of bounded salivary proteins and transmembrane mucin MUC1 expressed by oral epithelial cells. Previous research indicates that MUC1 expression enhances the binding of the main salivary protein forming the MP, MUC5B. This study investigated the influence of MUC1 structure on MP formation. A TR146 cell line, which does not express MUC1 natively, was stably transfected with genes coding for three MUC1 isoforms differing in the structure of the two main extracellular domains: the VNTR domain, exhibiting a variable number of tandem repeats, and the SEA domain, maintaining the two bound subunits of MUC1. Semi-quantification of MUC1 using dot blot chemiluminescence showed comparable expression levels in all transfected cell lines. Semi-quantification of MUC5B by immunostaining after incubation with saliva revealed that MUC1 expression significantly increased MUC5B adsorption. Neither the VNTR domain nor the SEA domain was influenced MUC5B anchoring, suggesting the key role of the MUC1 N-terminal domain. AFM-IR nanospectroscopy revealed discernible shifts indicative of changes in the chemical properties at the cell surface due to the expression of the MUC1 isoform. Furthermore, the observed chemical shifts suggest the involvement of hydrophobic effects in the interaction between MUC1 and salivary proteins.

AB - The mucosal pellicle (MP) is a biological film protecting the oral mucosa. It is composed of bounded salivary proteins and transmembrane mucin MUC1 expressed by oral epithelial cells. Previous research indicates that MUC1 expression enhances the binding of the main salivary protein forming the MP, MUC5B. This study investigated the influence of MUC1 structure on MP formation. A TR146 cell line, which does not express MUC1 natively, was stably transfected with genes coding for three MUC1 isoforms differing in the structure of the two main extracellular domains: the VNTR domain, exhibiting a variable number of tandem repeats, and the SEA domain, maintaining the two bound subunits of MUC1. Semi-quantification of MUC1 using dot blot chemiluminescence showed comparable expression levels in all transfected cell lines. Semi-quantification of MUC5B by immunostaining after incubation with saliva revealed that MUC1 expression significantly increased MUC5B adsorption. Neither the VNTR domain nor the SEA domain was influenced MUC5B anchoring, suggesting the key role of the MUC1 N-terminal domain. AFM-IR nanospectroscopy revealed discernible shifts indicative of changes in the chemical properties at the cell surface due to the expression of the MUC1 isoform. Furthermore, the observed chemical shifts suggest the involvement of hydrophobic effects in the interaction between MUC1 and salivary proteins.

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KW - MUC5B

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KW - oral mucosa

KW - salivary protein interactions

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Development of New Models of Oral Mucosa to Investigate the Impact of the Structure of Transmembrane Mucin-1 on the Mucosal Pellicle Formation and Its Physicochemical Properties

Abstract

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Keywords

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