The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations

Introduction: X-linked Alport syndrome (OMIM 301050) is caused by COL4A5 missense variants in 40% of families. This study examined the effects of chemical chaperone treatment (sodium 4-phenylbutyrate) on fibroblast cell lines derived from men with missense mutations. Methods: Dermal fibroblast cult...

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Main Authors:	Wang, Dongmao, Mohammad, Mardhiah, Wang, Yanyan, Tan, Rachel, Murray, Lydia S., Ricardo, Sharon, Dagher, Hayat, Agtmael, Tom van, Savige, Judy
Format:	Article
Language:	English
Published:	Elsevier 2017
Subjects:	Q Science (General) R Medicine (General)
Online Access:	http://irep.iium.edu.my/62219/ http://irep.iium.edu.my/62219/ http://irep.iium.edu.my/62219/ http://irep.iium.edu.my/62219/1/mardhiah2017.pdf

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spelling	iium-622192018-02-27T03:05:43Z http://irep.iium.edu.my/62219/ The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations Wang, Dongmao Mohammad, Mardhiah Wang, Yanyan Tan, Rachel Murray, Lydia S. Ricardo, Sharon Dagher, Hayat Agtmael, Tom van Savige, Judy Q Science (General) R Medicine (General) Introduction: X-linked Alport syndrome (OMIM 301050) is caused by COL4A5 missense variants in 40% of families. This study examined the effects of chemical chaperone treatment (sodium 4-phenylbutyrate) on fibroblast cell lines derived from men with missense mutations. Methods: Dermal fibroblast cultures were established from 2 affected men and 3 normals. Proliferation rates were examined, the collagen IV a5 chain localized with immunostaining, and levels of the intra- and extracellular chains quantitated with an in-house enzyme-linked immunosorbent assay. COL4A5 mRNA was measured using quantitative reverse transcriptase polymerase chain reaction. Endoplasmic reticulum (ER) size was measured on electron micrographs and after HSP47 immunostaining. Markers of ER stress (ATF6, HSPA5, DDIT3), autophagy (ATG5, BECN1, ATG7), and apoptosis (CASP3, BAD, BCL2) were also quantitated by quantitative reverse transcriptase polymerase chain reaction. Measurements were repeated after 48 hours of incubation with 10 mM sodium 4-phenylbutyrate acid. Results: Both COL4A5 missense variants were associated with reduced proliferation rates on day 6 (P 1⁄4 0.01 and P 1⁄4 0.03), ER enlargement, and increased mRNA for ER stress and autophagy (all P values < 0.05) when compared with normal. Sodium 4-phenylbutyrate treatment increased COL4A5 transcript levels (P < 0.01), and reduced ER size (P < 0.01 by EM and P < 0.001 by immunostaining), ER stress (p HSPA5 and DDIT3, all P values < 0.01) and autophagy (ATG7, P < 0.01). Extracellular collagen IV a5 chain was increased in the M1 line only (P 1⁄4 0.06). Discussion: Sodium 4-phenylbutyrate increases collagen IV a5 mRNA levels, reduces ER stress and autophagy, and possibly facilitates collagen IV a5 extracellular transport. Whether these actions delay end- stage renal failure in men with X-linked Alport syndrome and missense mutations will only be determined with clinical trials. Elsevier 2017-07 Article PeerReviewed application/pdf en http://irep.iium.edu.my/62219/1/mardhiah2017.pdf Wang, Dongmao and Mohammad, Mardhiah and Wang, Yanyan and Tan, Rachel and Murray, Lydia S. and Ricardo, Sharon and Dagher, Hayat and Agtmael, Tom van and Savige, Judy (2017) The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations. KI Reports - Kidney International Reports, 2 (4). pp. 739-748. ISSN 2468-0249 http://www.kireports.org/article/S2468-0249(17)30058-X/pdf http://dx.doi.org/10.1016/j.ekir.2017.03.004
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	Q Science (General) R Medicine (General)
spellingShingle	Q Science (General) R Medicine (General) Wang, Dongmao Mohammad, Mardhiah Wang, Yanyan Tan, Rachel Murray, Lydia S. Ricardo, Sharon Dagher, Hayat Agtmael, Tom van Savige, Judy The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations
description	Introduction: X-linked Alport syndrome (OMIM 301050) is caused by COL4A5 missense variants in 40% of families. This study examined the effects of chemical chaperone treatment (sodium 4-phenylbutyrate) on fibroblast cell lines derived from men with missense mutations. Methods: Dermal fibroblast cultures were established from 2 affected men and 3 normals. Proliferation rates were examined, the collagen IV a5 chain localized with immunostaining, and levels of the intra- and extracellular chains quantitated with an in-house enzyme-linked immunosorbent assay. COL4A5 mRNA was measured using quantitative reverse transcriptase polymerase chain reaction. Endoplasmic reticulum (ER) size was measured on electron micrographs and after HSP47 immunostaining. Markers of ER stress (ATF6, HSPA5, DDIT3), autophagy (ATG5, BECN1, ATG7), and apoptosis (CASP3, BAD, BCL2) were also quantitated by quantitative reverse transcriptase polymerase chain reaction. Measurements were repeated after 48 hours of incubation with 10 mM sodium 4-phenylbutyrate acid. Results: Both COL4A5 missense variants were associated with reduced proliferation rates on day 6 (P 1⁄4 0.01 and P 1⁄4 0.03), ER enlargement, and increased mRNA for ER stress and autophagy (all P values < 0.05) when compared with normal. Sodium 4-phenylbutyrate treatment increased COL4A5 transcript levels (P < 0.01), and reduced ER size (P < 0.01 by EM and P < 0.001 by immunostaining), ER stress (p HSPA5 and DDIT3, all P values < 0.01) and autophagy (ATG7, P < 0.01). Extracellular collagen IV a5 chain was increased in the M1 line only (P 1⁄4 0.06). Discussion: Sodium 4-phenylbutyrate increases collagen IV a5 mRNA levels, reduces ER stress and autophagy, and possibly facilitates collagen IV a5 extracellular transport. Whether these actions delay end- stage renal failure in men with X-linked Alport syndrome and missense mutations will only be determined with clinical trials.
format	Article
author	Wang, Dongmao Mohammad, Mardhiah Wang, Yanyan Tan, Rachel Murray, Lydia S. Ricardo, Sharon Dagher, Hayat Agtmael, Tom van Savige, Judy
author_facet	Wang, Dongmao Mohammad, Mardhiah Wang, Yanyan Tan, Rachel Murray, Lydia S. Ricardo, Sharon Dagher, Hayat Agtmael, Tom van Savige, Judy
author_sort	Wang, Dongmao
title	The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations
title_short	The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations
title_full	The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations
title_fullStr	The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations
title_full_unstemmed	The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations
title_sort	chemical chaperone, pba, reduces er stress and autophagy and increases collagen iv a5 expression in cultured fibroblasts from men with x-linked alport syndrome and missense mutations
publisher	Elsevier
publishDate	2017
url	http://irep.iium.edu.my/62219/ http://irep.iium.edu.my/62219/ http://irep.iium.edu.my/62219/ http://irep.iium.edu.my/62219/1/mardhiah2017.pdf
first_indexed	2023-09-18T21:28:12Z
last_indexed	2023-09-18T21:28:12Z
_version_	1777412334411579392

The chemical chaperone, PBA, reduces ER stress and autophagy and increases collagen IV a5 expression in cultured fibroblasts from men With X-linked alport syndrome and missense mutations

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