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Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro

BACKGROUND: Collagen is a structural protein that provides mechanical stability and defined architectures to skin. In collagen-based skin disorders this stability is lost, either due to mutations in collagens or in the chaperones involved in collagen assembly. This leads to chronic wounds, skin frag...

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Autores principales: Khan, Essak S., Sankaran, Shrikrishnan, Llontop, Lorena, del Campo, Aránzazu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7106624/
https://www.ncbi.nlm.nih.gov/pubmed/32228452
http://dx.doi.org/10.1186/s12860-020-00267-0
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author Khan, Essak S.
Sankaran, Shrikrishnan
Llontop, Lorena
del Campo, Aránzazu
author_facet Khan, Essak S.
Sankaran, Shrikrishnan
Llontop, Lorena
del Campo, Aránzazu
author_sort Khan, Essak S.
collection PubMed
description BACKGROUND: Collagen is a structural protein that provides mechanical stability and defined architectures to skin. In collagen-based skin disorders this stability is lost, either due to mutations in collagens or in the chaperones involved in collagen assembly. This leads to chronic wounds, skin fragility, and blistering. Existing approaches to treat such conditions rely on administration of small molecules to simulate collagen production, like 4-phenylbutyrate (4-PBA) or growth factors like TGF-β. However, these molecules are not specific for collagen synthesis, and result in unsolicited side effects. Hsp47 is a collagen-specific chaperone with a major role in collagen biosynthesis. Expression levels of Hsp47 correlate with collagen deposition. This article explores the stimulation of collagen deposition by exogenously supplied Hsp47 (collagen specific chaperone) to skin cells, including specific collagen subtypes quantification. RESULTS: Here we quantify the collagen deposition level and the types of deposited collagens after Hsp47 stimulation in different in vitro cultures of cells from human skin tissue (fibroblasts NHDF, keratinocytes HaCat and endothelial cells HDMEC) and mouse fibroblasts (L929 and MEF). We find upregulated deposition of fibrillar collagen subtypes I, III and V after Hsp47 delivery. Network collagen IV deposition was enhanced in HaCat and HDMECs, while fibril-associated collagen XII was not affected by the increased intracellular Hsp47 levels. The deposition levels of fibrillar collagen were cell-dependent i.e. Hsp47-stimulated fibroblasts deposited significantly higher amount of fibrillar collagen than Hsp47-stimulated HaCat and HDMECs. CONCLUSIONS: A 3-fold enhancement of collagen deposition was observed in fibroblasts upon repeated dosage of Hsp47 within the first 6 days of culture. Our results provide fundamental understanding towards the idea of using Hsp47 as therapeutic protein to treat collagen disorders.
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spelling pubmed-71066242020-04-01 Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro Khan, Essak S. Sankaran, Shrikrishnan Llontop, Lorena del Campo, Aránzazu BMC Mol Cell Biol Research Article BACKGROUND: Collagen is a structural protein that provides mechanical stability and defined architectures to skin. In collagen-based skin disorders this stability is lost, either due to mutations in collagens or in the chaperones involved in collagen assembly. This leads to chronic wounds, skin fragility, and blistering. Existing approaches to treat such conditions rely on administration of small molecules to simulate collagen production, like 4-phenylbutyrate (4-PBA) or growth factors like TGF-β. However, these molecules are not specific for collagen synthesis, and result in unsolicited side effects. Hsp47 is a collagen-specific chaperone with a major role in collagen biosynthesis. Expression levels of Hsp47 correlate with collagen deposition. This article explores the stimulation of collagen deposition by exogenously supplied Hsp47 (collagen specific chaperone) to skin cells, including specific collagen subtypes quantification. RESULTS: Here we quantify the collagen deposition level and the types of deposited collagens after Hsp47 stimulation in different in vitro cultures of cells from human skin tissue (fibroblasts NHDF, keratinocytes HaCat and endothelial cells HDMEC) and mouse fibroblasts (L929 and MEF). We find upregulated deposition of fibrillar collagen subtypes I, III and V after Hsp47 delivery. Network collagen IV deposition was enhanced in HaCat and HDMECs, while fibril-associated collagen XII was not affected by the increased intracellular Hsp47 levels. The deposition levels of fibrillar collagen were cell-dependent i.e. Hsp47-stimulated fibroblasts deposited significantly higher amount of fibrillar collagen than Hsp47-stimulated HaCat and HDMECs. CONCLUSIONS: A 3-fold enhancement of collagen deposition was observed in fibroblasts upon repeated dosage of Hsp47 within the first 6 days of culture. Our results provide fundamental understanding towards the idea of using Hsp47 as therapeutic protein to treat collagen disorders. BioMed Central 2020-03-30 /pmc/articles/PMC7106624/ /pubmed/32228452 http://dx.doi.org/10.1186/s12860-020-00267-0 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Khan, Essak S.
Sankaran, Shrikrishnan
Llontop, Lorena
del Campo, Aránzazu
Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro
title Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro
title_full Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro
title_fullStr Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro
title_full_unstemmed Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro
title_short Exogenous supply of Hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro
title_sort exogenous supply of hsp47 triggers fibrillar collagen deposition in skin cell cultures in vitro
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7106624/
https://www.ncbi.nlm.nih.gov/pubmed/32228452
http://dx.doi.org/10.1186/s12860-020-00267-0
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