Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.

Keratoconus (KC) is a complex thinning disease of the cornea that often requires transplantation. The underlying pathogenic molecular changes in this disease are poorly understood. Earlier studies reported oxidative stress, metabolic dysfunctions and accelerated death of stromal keratocytes in kerat...

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Autores principales:	James Foster, Wai-Hong Wu, Sherri-Gae Scott, Mehak Bassi, Divya Mohan, Yassine Daoud, Walter J Stark, Albert S Jun, Shukti Chakravarti
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f4dec07097db4acd9908f8d0a825204a
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spelling	oai:doaj.org-article:f4dec07097db4acd9908f8d0a825204a2021-11-25T05:59:45ZTransforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.1932-620310.1371/journal.pone.0106556https://doaj.org/article/f4dec07097db4acd9908f8d0a825204a2014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0106556https://doaj.org/toc/1932-6203Keratoconus (KC) is a complex thinning disease of the cornea that often requires transplantation. The underlying pathogenic molecular changes in this disease are poorly understood. Earlier studies reported oxidative stress, metabolic dysfunctions and accelerated death of stromal keratocytes in keratoconus (KC) patients. Utilizing mass spectrometry we found reduced stromal extracellular matrix (ECM) proteins in KC, suggesting ECM-regulatory changes that may be due to altered TGFβ signals. Here we investigated properties of stromal cells from donor (DN) and KC corneas grown as fibroblasts in serum containing DMEM: F12 or in serum-free medium containing insulin, transferrin, selenium (ITS). Phosphorylation of SMAD2/3 of the canonical TGFβ pathway, was high in serum-starved DN and KC fibroblast protein extracts, but pSMAD1/5/8 low at base line, was induced within 30 minutes of TGFβ1 stimulation, more so in KC than DN, suggesting a novel TGFβ1-SMAD1/5/8 axis in the cornea, that may be altered in KC. The serine/threonine kinases AKT, known to regulate proliferation, survival and biosynthetic activities of cells, were poorly activated in KC fibroblasts in high glucose media. Concordantly, alcohol dehydrogenase 1 (ADH1), an indicator of increased glucose uptake and metabolism, was reduced in KC compared to DN fibroblasts. By contrast, in low glucose (5.5 mM, normoglycemic) serum-free DMEM and ITS, cell survival and pAKT levels were comparable in KC and DN cells. Therefore, high glucose combined with serum-deprivation presents some cellular stress difficult to overcome by the KC stromal cells. Our study provides molecular insights into AKT and TGFβ signal changes in KC, and a mechanism for functional studies of stromal cells from KC corneas.James FosterWai-Hong WuSherri-Gae ScottMehak BassiDivya MohanYassine DaoudWalter J StarkAlbert S JunShukti ChakravartiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 9, p e106556 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q James Foster Wai-Hong Wu Sherri-Gae Scott Mehak Bassi Divya Mohan Yassine Daoud Walter J Stark Albert S Jun Shukti Chakravarti Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.
description	Keratoconus (KC) is a complex thinning disease of the cornea that often requires transplantation. The underlying pathogenic molecular changes in this disease are poorly understood. Earlier studies reported oxidative stress, metabolic dysfunctions and accelerated death of stromal keratocytes in keratoconus (KC) patients. Utilizing mass spectrometry we found reduced stromal extracellular matrix (ECM) proteins in KC, suggesting ECM-regulatory changes that may be due to altered TGFβ signals. Here we investigated properties of stromal cells from donor (DN) and KC corneas grown as fibroblasts in serum containing DMEM: F12 or in serum-free medium containing insulin, transferrin, selenium (ITS). Phosphorylation of SMAD2/3 of the canonical TGFβ pathway, was high in serum-starved DN and KC fibroblast protein extracts, but pSMAD1/5/8 low at base line, was induced within 30 minutes of TGFβ1 stimulation, more so in KC than DN, suggesting a novel TGFβ1-SMAD1/5/8 axis in the cornea, that may be altered in KC. The serine/threonine kinases AKT, known to regulate proliferation, survival and biosynthetic activities of cells, were poorly activated in KC fibroblasts in high glucose media. Concordantly, alcohol dehydrogenase 1 (ADH1), an indicator of increased glucose uptake and metabolism, was reduced in KC compared to DN fibroblasts. By contrast, in low glucose (5.5 mM, normoglycemic) serum-free DMEM and ITS, cell survival and pAKT levels were comparable in KC and DN cells. Therefore, high glucose combined with serum-deprivation presents some cellular stress difficult to overcome by the KC stromal cells. Our study provides molecular insights into AKT and TGFβ signal changes in KC, and a mechanism for functional studies of stromal cells from KC corneas.
format	article
author	James Foster Wai-Hong Wu Sherri-Gae Scott Mehak Bassi Divya Mohan Yassine Daoud Walter J Stark Albert S Jun Shukti Chakravarti
author_facet	James Foster Wai-Hong Wu Sherri-Gae Scott Mehak Bassi Divya Mohan Yassine Daoud Walter J Stark Albert S Jun Shukti Chakravarti
author_sort	James Foster
title	Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.
title_short	Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.
title_full	Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.
title_fullStr	Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.
title_full_unstemmed	Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.
title_sort	transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/f4dec07097db4acd9908f8d0a825204a
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Transforming growth factor β and insulin signal changes in stromal fibroblasts of individual keratoconus patients.

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