FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress

Abstract Retinal pigment epithelial (RPE) cells that underlie the neurosensory retina are essential for the maintenance of photoreceptor cells and hence vision. Interactions between the RPE and their basement membrane, i.e. the inner layer of Bruch’s membrane, are essential for RPE cell health and f...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Rawshan Choudhury, Nadhim Bayatti, Richard Scharff, Ewa Szula, Viranga Tilakaratna, Maja Søberg Udsen, Selina McHarg, Janet A. Askari, Martin J. Humphries, Paul N. Bishop, Simon J. Clark
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/744e5ccb41c64bcf9d0d0a1eaadb8a8c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:744e5ccb41c64bcf9d0d0a1eaadb8a8c
record_format	dspace
spelling	oai:doaj.org-article:744e5ccb41c64bcf9d0d0a1eaadb8a8c2021-12-02T16:15:06ZFHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress10.1038/s41598-021-93708-52045-2322https://doaj.org/article/744e5ccb41c64bcf9d0d0a1eaadb8a8c2021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-93708-5https://doaj.org/toc/2045-2322Abstract Retinal pigment epithelial (RPE) cells that underlie the neurosensory retina are essential for the maintenance of photoreceptor cells and hence vision. Interactions between the RPE and their basement membrane, i.e. the inner layer of Bruch’s membrane, are essential for RPE cell health and function, but the signals induced by Bruch’s membrane engagement, and their contributions to RPE cell fate determination remain poorly defined. Here, we studied the functional role of the soluble complement regulator and component of Bruch’s membrane, Factor H-like protein 1 (FHL-1). Human primary RPE cells adhered to FHL-1 in a manner that was eliminated by either mutagenesis of the integrin-binding RGD motif in FHL-1 or by using competing antibodies directed against the α5 and β1 integrin subunits. These short-term experiments reveal an immediate protein-integrin interaction that were obtained from primary RPE cells and replicated using the hTERT-RPE1 cell line. Separate, longer term experiments utilising RNAseq analysis of hTERT-RPE1 cells bound to FHL-1, showed an increased expression of the heat-shock protein genes HSPA6, CRYAB, HSPA1A and HSPA1B when compared to cells bound to fibronectin (FN) or laminin (LA). Pathway analysis implicated changes in EIF2 signalling, the unfolded protein response, and mineralocorticoid receptor signalling as putative pathways. Subsequent cell survival assays using H2O2 to induce oxidative stress-induced cell death suggest hTERT-RPE1 cells had significantly greater protection when bound to FHL-1 or LA compared to plastic or FN. These data show a non-canonical role of FHL-1 in protecting RPE cells against oxidative stress and identifies a novel interaction that has implications for ocular diseases such as age-related macular degeneration.Rawshan ChoudhuryNadhim BayattiRichard ScharffEwa SzulaViranga TilakaratnaMaja Søberg UdsenSelina McHargJanet A. AskariMartin J. HumphriesPaul N. BishopSimon J. ClarkNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-17 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rawshan Choudhury Nadhim Bayatti Richard Scharff Ewa Szula Viranga Tilakaratna Maja Søberg Udsen Selina McHarg Janet A. Askari Martin J. Humphries Paul N. Bishop Simon J. Clark FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress
description	Abstract Retinal pigment epithelial (RPE) cells that underlie the neurosensory retina are essential for the maintenance of photoreceptor cells and hence vision. Interactions between the RPE and their basement membrane, i.e. the inner layer of Bruch’s membrane, are essential for RPE cell health and function, but the signals induced by Bruch’s membrane engagement, and their contributions to RPE cell fate determination remain poorly defined. Here, we studied the functional role of the soluble complement regulator and component of Bruch’s membrane, Factor H-like protein 1 (FHL-1). Human primary RPE cells adhered to FHL-1 in a manner that was eliminated by either mutagenesis of the integrin-binding RGD motif in FHL-1 or by using competing antibodies directed against the α5 and β1 integrin subunits. These short-term experiments reveal an immediate protein-integrin interaction that were obtained from primary RPE cells and replicated using the hTERT-RPE1 cell line. Separate, longer term experiments utilising RNAseq analysis of hTERT-RPE1 cells bound to FHL-1, showed an increased expression of the heat-shock protein genes HSPA6, CRYAB, HSPA1A and HSPA1B when compared to cells bound to fibronectin (FN) or laminin (LA). Pathway analysis implicated changes in EIF2 signalling, the unfolded protein response, and mineralocorticoid receptor signalling as putative pathways. Subsequent cell survival assays using H2O2 to induce oxidative stress-induced cell death suggest hTERT-RPE1 cells had significantly greater protection when bound to FHL-1 or LA compared to plastic or FN. These data show a non-canonical role of FHL-1 in protecting RPE cells against oxidative stress and identifies a novel interaction that has implications for ocular diseases such as age-related macular degeneration.
format	article
author	Rawshan Choudhury Nadhim Bayatti Richard Scharff Ewa Szula Viranga Tilakaratna Maja Søberg Udsen Selina McHarg Janet A. Askari Martin J. Humphries Paul N. Bishop Simon J. Clark
author_facet	Rawshan Choudhury Nadhim Bayatti Richard Scharff Ewa Szula Viranga Tilakaratna Maja Søberg Udsen Selina McHarg Janet A. Askari Martin J. Humphries Paul N. Bishop Simon J. Clark
author_sort	Rawshan Choudhury
title	FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress
title_short	FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress
title_full	FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress
title_fullStr	FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress
title_full_unstemmed	FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress
title_sort	fhl-1 interacts with human rpe cells through the α5β1 integrin and confers protection against oxidative stress
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/744e5ccb41c64bcf9d0d0a1eaadb8a8c
work_keys_str_mv	AT rawshanchoudhury fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT nadhimbayatti fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT richardscharff fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT ewaszula fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT virangatilakaratna fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT majasøbergudsen fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT selinamcharg fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT janetaaskari fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT martinjhumphries fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT paulnbishop fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress AT simonjclark fhl1interactswithhumanrpecellsthroughthea5b1integrinandconfersprotectionagainstoxidativestress
_version_	1718384297723822080

FHL-1 interacts with human RPE cells through the α5β1 integrin and confers protection against oxidative stress

Ejemplares similares