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Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs

HSP-SPAST is the most common form of hereditary spastic paraplegia (HSP), a neurodegenerative disease causing lower limb spasticity. Previous studies using HSP-SPAST patient-derived induced pluripotent stem cell cortical neurons have shown that patient neurons have reduced levels of acetylated α-tub...

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Autores principales: Wali, Gautam, Siow, Sue-Faye, Liyanage, Erandhi, Kumar, Kishore R., Mackay-Sim, Alan, Sue, Carolyn M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10152469/
https://www.ncbi.nlm.nih.gov/pubmed/37144097
http://dx.doi.org/10.3389/fnins.2023.1073516
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author Wali, Gautam
Siow, Sue-Faye
Liyanage, Erandhi
Kumar, Kishore R.
Mackay-Sim, Alan
Sue, Carolyn M.
author_facet Wali, Gautam
Siow, Sue-Faye
Liyanage, Erandhi
Kumar, Kishore R.
Mackay-Sim, Alan
Sue, Carolyn M.
author_sort Wali, Gautam
collection PubMed
description HSP-SPAST is the most common form of hereditary spastic paraplegia (HSP), a neurodegenerative disease causing lower limb spasticity. Previous studies using HSP-SPAST patient-derived induced pluripotent stem cell cortical neurons have shown that patient neurons have reduced levels of acetylated α-tubulin, a form of stabilized microtubules, leading to a chain of downstream effects eventuating in increased vulnerability to axonal degeneration. Noscapine treatment rescued these downstream effects by restoring the levels of acetylated α-tubulin in patient neurons. Here we show that HSP-SPAST patient non-neuronal cells, peripheral blood mononuclear cells (PBMCs), also have the disease-associated effect of reduced levels of acetylated α-tubulin. Evaluation of multiple PBMC subtypes showed that patient T cell lymphocytes had reduced levels of acetylated α-tubulin. T cells make up to 80% of all PBMCs and likely contributed to the effect of reduced acetylated α-tubulin levels seen in overall PBMCs. We further showed that mouse administered orally with increasing concentrations of noscapine exhibited a dose-dependent increase of noscapine levels and acetylated α-tubulin in the brain. A similar effect of noscapine treatment is anticipated in HSP-SPAST patients. To measure acetylated α-tubulin levels, we used a homogeneous time resolved fluorescence technology-based assay. This assay was sensitive to noscapine-induced changes in acetylated α-tubulin levels in multiple sample types. The assay is high throughput and uses nano-molar protein concentrations, making it an ideal assay for evaluation of noscapine-induced changes in acetylated α-tubulin levels. This study shows that HSP-SPAST patient PBMCs exhibit disease-associated effects. This finding can help expedite the drug discovery and testing process.
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spelling pubmed-101524692023-05-03 Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs Wali, Gautam Siow, Sue-Faye Liyanage, Erandhi Kumar, Kishore R. Mackay-Sim, Alan Sue, Carolyn M. Front Neurosci Neuroscience HSP-SPAST is the most common form of hereditary spastic paraplegia (HSP), a neurodegenerative disease causing lower limb spasticity. Previous studies using HSP-SPAST patient-derived induced pluripotent stem cell cortical neurons have shown that patient neurons have reduced levels of acetylated α-tubulin, a form of stabilized microtubules, leading to a chain of downstream effects eventuating in increased vulnerability to axonal degeneration. Noscapine treatment rescued these downstream effects by restoring the levels of acetylated α-tubulin in patient neurons. Here we show that HSP-SPAST patient non-neuronal cells, peripheral blood mononuclear cells (PBMCs), also have the disease-associated effect of reduced levels of acetylated α-tubulin. Evaluation of multiple PBMC subtypes showed that patient T cell lymphocytes had reduced levels of acetylated α-tubulin. T cells make up to 80% of all PBMCs and likely contributed to the effect of reduced acetylated α-tubulin levels seen in overall PBMCs. We further showed that mouse administered orally with increasing concentrations of noscapine exhibited a dose-dependent increase of noscapine levels and acetylated α-tubulin in the brain. A similar effect of noscapine treatment is anticipated in HSP-SPAST patients. To measure acetylated α-tubulin levels, we used a homogeneous time resolved fluorescence technology-based assay. This assay was sensitive to noscapine-induced changes in acetylated α-tubulin levels in multiple sample types. The assay is high throughput and uses nano-molar protein concentrations, making it an ideal assay for evaluation of noscapine-induced changes in acetylated α-tubulin levels. This study shows that HSP-SPAST patient PBMCs exhibit disease-associated effects. This finding can help expedite the drug discovery and testing process. Frontiers Media S.A. 2023-03-28 /pmc/articles/PMC10152469/ /pubmed/37144097 http://dx.doi.org/10.3389/fnins.2023.1073516 Text en Copyright © 2023 Wali, Siow, Liyanage, Kumar, Mackay-Sim and Sue. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
Wali, Gautam
Siow, Sue-Faye
Liyanage, Erandhi
Kumar, Kishore R.
Mackay-Sim, Alan
Sue, Carolyn M.
Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs
title Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs
title_full Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs
title_fullStr Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs
title_full_unstemmed Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs
title_short Reduced acetylated α-tubulin in SPAST hereditary spastic paraplegia patient PBMCs
title_sort reduced acetylated α-tubulin in spast hereditary spastic paraplegia patient pbmcs
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10152469/
https://www.ncbi.nlm.nih.gov/pubmed/37144097
http://dx.doi.org/10.3389/fnins.2023.1073516
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