Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas

In order to reduce the need for donor corneas, understanding of corneal wound healing and development of an entirely tissue-engineered human cornea (hTECs) is of prime importance. In this study, we exploited the hTEC to determine how deep wound healing affects the transcriptional pattern of corneal...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Christelle Gross, Gaëtan Le-Bel, Pascale Desjardins, Manel Benhassine, Lucie Germain, Sylvain L. Guérin
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>clusterin</i> human cornea tissue-engineering 3D corneal model wound healing transcription factor Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/0dc8252090374a42bb40e44f4ce68fb9
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:0dc8252090374a42bb40e44f4ce68fb9
record_format	dspace
spelling	oai:doaj.org-article:0dc8252090374a42bb40e44f4ce68fb92021-11-25T17:56:32ZContribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas10.3390/ijms2222124261422-00671661-6596https://doaj.org/article/0dc8252090374a42bb40e44f4ce68fb92021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12426https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067In order to reduce the need for donor corneas, understanding of corneal wound healing and development of an entirely tissue-engineered human cornea (hTECs) is of prime importance. In this study, we exploited the hTEC to determine how deep wound healing affects the transcriptional pattern of corneal epithelial cells through microarray analyses. We demonstrated that the gene encoding <i>clusterin</i> (<i>CLU</i>) has its expression dramatically repressed during closure of hTEC wounds. Western blot analyses confirmed a strong reduction in the expression of the <i>clusterin</i> isoforms after corneal damage and suggest that repression of <i>CLU</i> gene expression might be a prerequisite to hTEC wound closure. Transfection with segments from the human <i>CLU</i> gene promoter revealed the presence of three regulatory regions: a basal promoter and two more distal negative regulatory regions. The basal promoter bears DNA binding sites for very potent transcription factors (TFs): Activator Protein-1 (AP-1) and Specificity protein-1 and 3 (Sp1/Sp3). By exploiting electrophoretic mobility shift assays (EMSA), we demonstrated that AP-1 and Sp1/Sp3 have their DNA binding site overlapping with one another in the basal promoter of the <i>CLU</i> gene in hCECs. Interestingly, expression of both these TFs is reduced (at the protein level) during hTEC wound healing, thereby contributing to the extinction of <i>CLU</i> gene expression during that process. The results of this study contribute to a better understanding of the molecular mechanisms accounting for the repression of <i>CLU</i> gene expression during corneal wound healing.Christelle GrossGaëtan Le-BelPascale DesjardinsManel BenhassineLucie GermainSylvain L. GuérinMDPI AGarticle<i>clusterin</i>human corneatissue-engineering3D corneal modelwound healingtranscription factorBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12426, p 12426 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>clusterin</i> human cornea tissue-engineering 3D corneal model wound healing transcription factor Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	<i>clusterin</i> human cornea tissue-engineering 3D corneal model wound healing transcription factor Biology (General) QH301-705.5 Chemistry QD1-999 Christelle Gross Gaëtan Le-Bel Pascale Desjardins Manel Benhassine Lucie Germain Sylvain L. Guérin Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas
description	In order to reduce the need for donor corneas, understanding of corneal wound healing and development of an entirely tissue-engineered human cornea (hTECs) is of prime importance. In this study, we exploited the hTEC to determine how deep wound healing affects the transcriptional pattern of corneal epithelial cells through microarray analyses. We demonstrated that the gene encoding <i>clusterin</i> (<i>CLU</i>) has its expression dramatically repressed during closure of hTEC wounds. Western blot analyses confirmed a strong reduction in the expression of the <i>clusterin</i> isoforms after corneal damage and suggest that repression of <i>CLU</i> gene expression might be a prerequisite to hTEC wound closure. Transfection with segments from the human <i>CLU</i> gene promoter revealed the presence of three regulatory regions: a basal promoter and two more distal negative regulatory regions. The basal promoter bears DNA binding sites for very potent transcription factors (TFs): Activator Protein-1 (AP-1) and Specificity protein-1 and 3 (Sp1/Sp3). By exploiting electrophoretic mobility shift assays (EMSA), we demonstrated that AP-1 and Sp1/Sp3 have their DNA binding site overlapping with one another in the basal promoter of the <i>CLU</i> gene in hCECs. Interestingly, expression of both these TFs is reduced (at the protein level) during hTEC wound healing, thereby contributing to the extinction of <i>CLU</i> gene expression during that process. The results of this study contribute to a better understanding of the molecular mechanisms accounting for the repression of <i>CLU</i> gene expression during corneal wound healing.
format	article
author	Christelle Gross Gaëtan Le-Bel Pascale Desjardins Manel Benhassine Lucie Germain Sylvain L. Guérin
author_facet	Christelle Gross Gaëtan Le-Bel Pascale Desjardins Manel Benhassine Lucie Germain Sylvain L. Guérin
author_sort	Christelle Gross
title	Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas
title_short	Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas
title_full	Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas
title_fullStr	Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas
title_full_unstemmed	Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas
title_sort	contribution of the transcription factors sp1/sp3 and ap-1 to <i>clusterin</i> gene expression during corneal wound healing of tissue-engineered human corneas
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/0dc8252090374a42bb40e44f4ce68fb9
work_keys_str_mv	AT christellegross contributionofthetranscriptionfactorssp1sp3andap1toiclusterinigeneexpressionduringcornealwoundhealingoftissueengineeredhumancorneas AT gaetanlebel contributionofthetranscriptionfactorssp1sp3andap1toiclusterinigeneexpressionduringcornealwoundhealingoftissueengineeredhumancorneas AT pascaledesjardins contributionofthetranscriptionfactorssp1sp3andap1toiclusterinigeneexpressionduringcornealwoundhealingoftissueengineeredhumancorneas AT manelbenhassine contributionofthetranscriptionfactorssp1sp3andap1toiclusterinigeneexpressionduringcornealwoundhealingoftissueengineeredhumancorneas AT luciegermain contributionofthetranscriptionfactorssp1sp3andap1toiclusterinigeneexpressionduringcornealwoundhealingoftissueengineeredhumancorneas AT sylvainlguerin contributionofthetranscriptionfactorssp1sp3andap1toiclusterinigeneexpressionduringcornealwoundhealingoftissueengineeredhumancorneas
_version_	1718411784952479744

Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to <i>Clusterin</i> Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas

Ejemplares similares