Cargando…

Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease

Fabry disease stems from a deficiency of alpha-galactosidase and results in the accumulation of globotriaosylceramide (Gb3). However, the production of its deacylated form globotriaosylsphingosine (lyso-Gb3) is also observed and its plasma levels have closer association with disease severity. Studie...

Descripción completa

Detalles Bibliográficos
Autores principales: Nikolaenko, Valeria, Warnock, David G, Mills, Kevin, Heywood, Wendy E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10360391/
https://www.ncbi.nlm.nih.gov/pubmed/37145097
http://dx.doi.org/10.1093/hmg/ddad073
_version_ 1785076094473338880
author Nikolaenko, Valeria
Warnock, David G
Mills, Kevin
Heywood, Wendy E
author_facet Nikolaenko, Valeria
Warnock, David G
Mills, Kevin
Heywood, Wendy E
author_sort Nikolaenko, Valeria
collection PubMed
description Fabry disease stems from a deficiency of alpha-galactosidase and results in the accumulation of globotriaosylceramide (Gb3). However, the production of its deacylated form globotriaosylsphingosine (lyso-Gb3) is also observed and its plasma levels have closer association with disease severity. Studies have shown that lyso-Gb3 directly affects podocytes and causes sensitisation of peripheral nociceptive neurons. However, little is understood of the mechanisms of this cytotoxicity. To study the effect on neuronal cells, we incubated SH-Sy5y cells with lyso-Gb3 at low (20 ng/mL) and high (200 ng/mL) levels, to mimic mild and classical FD serum levels. We used glucosylsphingosine as a positive control to determine specific effects of lyso-Gb3. Proteomic analyses revealed that cellular systems affected by lyso-Gb3 included cell signalling particularly protein ubiquitination and protein translation. To confirm ER/proteasome perturbations, we performed an immune enrichment of ubiquitinated proteins and demonstrated specific increased protein ubiquitination at both doses. The most ubiquitinated proteins observed included the chaperone/heat shock proteins, cytoskeletal proteins and synthesis/translation proteins. To detect proteins that interact directly with lyso-Gb3, we immobilised lyso-lipids, then incubated them with neuronal cellular extracts and identified bound proteins using mass spectrometry. Proteins that specifically bound were chaperones and included HSP90, HSP60 and the TRiC complex. In conclusion, lyso-Gb3 exposure affects pathways involved in protein translation and folding. This response is observed as increased ubiquitination and changes in signalling proteins which may explain the multiple biological processes, particularly cellular remodelling, often associated with FD.
format Online
Article
Text
id pubmed-10360391
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-103603912023-07-22 Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease Nikolaenko, Valeria Warnock, David G Mills, Kevin Heywood, Wendy E Hum Mol Genet Original Article Fabry disease stems from a deficiency of alpha-galactosidase and results in the accumulation of globotriaosylceramide (Gb3). However, the production of its deacylated form globotriaosylsphingosine (lyso-Gb3) is also observed and its plasma levels have closer association with disease severity. Studies have shown that lyso-Gb3 directly affects podocytes and causes sensitisation of peripheral nociceptive neurons. However, little is understood of the mechanisms of this cytotoxicity. To study the effect on neuronal cells, we incubated SH-Sy5y cells with lyso-Gb3 at low (20 ng/mL) and high (200 ng/mL) levels, to mimic mild and classical FD serum levels. We used glucosylsphingosine as a positive control to determine specific effects of lyso-Gb3. Proteomic analyses revealed that cellular systems affected by lyso-Gb3 included cell signalling particularly protein ubiquitination and protein translation. To confirm ER/proteasome perturbations, we performed an immune enrichment of ubiquitinated proteins and demonstrated specific increased protein ubiquitination at both doses. The most ubiquitinated proteins observed included the chaperone/heat shock proteins, cytoskeletal proteins and synthesis/translation proteins. To detect proteins that interact directly with lyso-Gb3, we immobilised lyso-lipids, then incubated them with neuronal cellular extracts and identified bound proteins using mass spectrometry. Proteins that specifically bound were chaperones and included HSP90, HSP60 and the TRiC complex. In conclusion, lyso-Gb3 exposure affects pathways involved in protein translation and folding. This response is observed as increased ubiquitination and changes in signalling proteins which may explain the multiple biological processes, particularly cellular remodelling, often associated with FD. Oxford University Press 2023-05-05 /pmc/articles/PMC10360391/ /pubmed/37145097 http://dx.doi.org/10.1093/hmg/ddad073 Text en © The Author(s) 2023. Published by Oxford University Press. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Nikolaenko, Valeria
Warnock, David G
Mills, Kevin
Heywood, Wendy E
Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease
title Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease
title_full Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease
title_fullStr Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease
title_full_unstemmed Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease
title_short Elucidating the toxic effect and disease mechanisms associated with Lyso-Gb3 in Fabry disease
title_sort elucidating the toxic effect and disease mechanisms associated with lyso-gb3 in fabry disease
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10360391/
https://www.ncbi.nlm.nih.gov/pubmed/37145097
http://dx.doi.org/10.1093/hmg/ddad073
work_keys_str_mv AT nikolaenkovaleria elucidatingthetoxiceffectanddiseasemechanismsassociatedwithlysogb3infabrydisease
AT warnockdavidg elucidatingthetoxiceffectanddiseasemechanismsassociatedwithlysogb3infabrydisease
AT millskevin elucidatingthetoxiceffectanddiseasemechanismsassociatedwithlysogb3infabrydisease
AT heywoodwendye elucidatingthetoxiceffectanddiseasemechanismsassociatedwithlysogb3infabrydisease