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Inhibition of PHLDA3 expression in human superoxide dismutase 1-mutant amyotrophic lateral sclerosis astrocytes protects against neurotoxicity.

Published version
Peer-reviewed

Repository DOI


Change log

Authors

Szebényi, Kornélia 
Vargová, Ingrid 
Turečková, Jana 
Gibbons, George M 

Abstract

Pleckstrin homology-like domain family A-member 3 (PHLDA3) has recently been identified as a player in adaptive and maladaptive cellular stress pathways. The outcome of pleckstrin homology-like domain family A-member 3 signalling was shown to vary across different cell types and states. It emerges that its expression and protein level are highly increased in amyotrophic lateral sclerosis (ALS) patient-derived astrocytes. Whether it orchestrates a supportive or detrimental function remains unexplored in the context of neurodegenerative pathologies. To directly address the role of pleckstrin homology-like domain family A-member 3 in healthy and ALS astrocytes, we used overexpression and knockdown strategies. We generated cultures of primary mouse astrocytes and also human astrocytes from control and ALS patient-derived induced pluripotent stem cells harbouring the superoxide dismutase 1 mutation. Then, we assessed astrocyte viability and the impact of their secretome on oxidative stress responses in human stem cell-derived cortical and spinal neuronal cultures. Here, we show that PHLDA3 overexpression or knockdown in control astrocytes does not significantly affect astrocyte viability or reactive oxygen species production. However, PHLDA3 knockdown in ALS astrocytes diminishes reactive oxygen species concentrations in their supernatants, indicating that pleckstrin homology-like domain family A-member 3 can facilitate stress responses in cells with altered homeostasis. In support, supernatants of PHLDA3-silenced ALS and even control spinal astrocytes with a lower pleckstrin homology-like domain family A-member 3 protein content could prevent sodium arsenite-induced stress granule formation in spinal neurons. Our findings provide evidence that reducing pleckstrin homology-like domain family A-member 3 levels may transform astrocytes into a more neurosupportive state relevant to targeting non-cell autonomous ALS pathology.

Description

Acknowledgements: The authors thank Mr Damion Box for his technical support.


Funder: International Brain Research Organisation; doi: https://doi.org/10.13039/501100001675


Funder: National Research, Development and Innovation Fund; doi: https://doi.org/10.13039/501100012550


Funder: NeuroRecon Project

Keywords

PHLDA3, amyotrophic lateral sclerosis, astrocyte, astrocyte–neuron interaction, cell stress

Journal Title

Brain Commun

Conference Name

Journal ISSN

2632-1297
2632-1297

Volume Title

6

Publisher

Oxford University Press (OUP)
Sponsorship
MRC (MR/X006867/1)
Medical Research Council (MR/P008658/1)
This project and the A.L. laboratory were funded by the Medical Research Council UK (United Kingdom Research and Innovation, Medical Research Council, MR/P008658/1 and MR/X006867/1 to A.L.), which supported the work of K.SZ., G.M.G. and V.P. Additional support was received from the International Brain Research Organisation Return Home Fellowship 2022, the National Research, Development and Innovation Fund/Hungarian Scientific Research Fund (NKFIH/OTKA FK 142223 for K.SZ., A.S.), and the Grant Agency of the Czech Republic (GAC¬R 17-21146 S) and the NeuroRecon Project (Operational Programme Research, Development and Education in the framework of the project “Center of Reconstructive Neuroscience,” registration number CZ.02.1.01/0.0./0.0/15_003/0000419 to P.J., J.C.F.K. and J.W.F.) for the work of I.V., J.T., and M.S.