Regulation of retinal pigment epithelial cell phenotype by Annexin A8

Abstract The retinoic acid derivative fenretinide (FR) is capable of transdifferentiating cultured retinal pigment epithelial (RPE) cells towards a neuronal-like phenotype, but the underlying mechanisms are not understood. To identify genes involved in this process we performed a microarray analysis...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Katharina Lueck, Amanda-Jayne F. Carr, Dimitrios Stampoulis, Volker Gerke, Ursula Rescher, John Greenwood, Stephen E. Moss
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/51d55e5e3165498991da240361a0634b
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:51d55e5e3165498991da240361a0634b
record_format	dspace
spelling	oai:doaj.org-article:51d55e5e3165498991da240361a0634b2021-12-02T15:04:53ZRegulation of retinal pigment epithelial cell phenotype by Annexin A810.1038/s41598-017-03493-32045-2322https://doaj.org/article/51d55e5e3165498991da240361a0634b2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-03493-3https://doaj.org/toc/2045-2322Abstract The retinoic acid derivative fenretinide (FR) is capable of transdifferentiating cultured retinal pigment epithelial (RPE) cells towards a neuronal-like phenotype, but the underlying mechanisms are not understood. To identify genes involved in this process we performed a microarray analysis of RPE cells pre- and post-FR treatment, and observed a marked down-regulation of AnnexinA8 (AnxA8) in transdifferentiated cells. To determine whether AnxA8 plays a role in maintaining RPE cell phenotype we directly manipulated AnxA8 expression in cultured and primary RPE cells using siRNA-mediated gene suppression, and over-expression of AnxA8-GFP in conjunction with exposure to FR. Treatment of RPE cells with AnxA8 siRNA recapitulated exposure to FR, with cell cycle arrest, neuronal transdifferentiation, and concomitant up-regulation of the neuronal markers calretinin and calbindin, as assessed by real-time PCR and immunofluorescence. In contrast, AnxA8 transient over-expression in ARPE-19 cells prevented FR-induced differentiation. Ectopic expression of AnxA8 in AnxA8-depleted cells led to decreased neuronal marker staining, and normal cell growth as judged by phosphohistone H3 staining, cell counting and cleaved caspase-3 levels. These data show that down-regulation of AnxA8 is both necessary and sufficient for neuronal transdifferentiation of RPE cells and reveal an essential role for AnxA8 as a key regulator of RPE phenotype.Katharina LueckAmanda-Jayne F. CarrDimitrios StampoulisVolker GerkeUrsula RescherJohn GreenwoodStephen E. MossNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Katharina Lueck Amanda-Jayne F. Carr Dimitrios Stampoulis Volker Gerke Ursula Rescher John Greenwood Stephen E. Moss Regulation of retinal pigment epithelial cell phenotype by Annexin A8
description	Abstract The retinoic acid derivative fenretinide (FR) is capable of transdifferentiating cultured retinal pigment epithelial (RPE) cells towards a neuronal-like phenotype, but the underlying mechanisms are not understood. To identify genes involved in this process we performed a microarray analysis of RPE cells pre- and post-FR treatment, and observed a marked down-regulation of AnnexinA8 (AnxA8) in transdifferentiated cells. To determine whether AnxA8 plays a role in maintaining RPE cell phenotype we directly manipulated AnxA8 expression in cultured and primary RPE cells using siRNA-mediated gene suppression, and over-expression of AnxA8-GFP in conjunction with exposure to FR. Treatment of RPE cells with AnxA8 siRNA recapitulated exposure to FR, with cell cycle arrest, neuronal transdifferentiation, and concomitant up-regulation of the neuronal markers calretinin and calbindin, as assessed by real-time PCR and immunofluorescence. In contrast, AnxA8 transient over-expression in ARPE-19 cells prevented FR-induced differentiation. Ectopic expression of AnxA8 in AnxA8-depleted cells led to decreased neuronal marker staining, and normal cell growth as judged by phosphohistone H3 staining, cell counting and cleaved caspase-3 levels. These data show that down-regulation of AnxA8 is both necessary and sufficient for neuronal transdifferentiation of RPE cells and reveal an essential role for AnxA8 as a key regulator of RPE phenotype.
format	article
author	Katharina Lueck Amanda-Jayne F. Carr Dimitrios Stampoulis Volker Gerke Ursula Rescher John Greenwood Stephen E. Moss
author_facet	Katharina Lueck Amanda-Jayne F. Carr Dimitrios Stampoulis Volker Gerke Ursula Rescher John Greenwood Stephen E. Moss
author_sort	Katharina Lueck
title	Regulation of retinal pigment epithelial cell phenotype by Annexin A8
title_short	Regulation of retinal pigment epithelial cell phenotype by Annexin A8
title_full	Regulation of retinal pigment epithelial cell phenotype by Annexin A8
title_fullStr	Regulation of retinal pigment epithelial cell phenotype by Annexin A8
title_full_unstemmed	Regulation of retinal pigment epithelial cell phenotype by Annexin A8
title_sort	regulation of retinal pigment epithelial cell phenotype by annexin a8
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/51d55e5e3165498991da240361a0634b
work_keys_str_mv	AT katharinalueck regulationofretinalpigmentepithelialcellphenotypebyannexina8 AT amandajaynefcarr regulationofretinalpigmentepithelialcellphenotypebyannexina8 AT dimitriosstampoulis regulationofretinalpigmentepithelialcellphenotypebyannexina8 AT volkergerke regulationofretinalpigmentepithelialcellphenotypebyannexina8 AT ursularescher regulationofretinalpigmentepithelialcellphenotypebyannexina8 AT johngreenwood regulationofretinalpigmentepithelialcellphenotypebyannexina8 AT stephenemoss regulationofretinalpigmentepithelialcellphenotypebyannexina8
_version_	1718389015521001472

Regulation of retinal pigment epithelial cell phenotype by Annexin A8

Ejemplares similares