Transactive response DNA-binding protein-43 proteinopathy in oligodendrocytes revealed using an induced pluripotent stem cell model

Samantha K Barton; Dario Magnani; Owen G James; Matthew R Livesey; Bhuvaneish T Selvaraj; Owain T James; Emma M Perkins; Jenna M Gregory; Elaine Cleary; C Rosanne M Ausems; Roderick N Carter Carter; Navneet A Vasistha; Chen Zhao; Karen Burr; David Story; Alessandra Cardinali; Nicholas M Morton; Giles E Hardingham; David J A Wyllie; Siddharthan Chandran

doi:10.1093/braincomms/fcab255

Transactive response DNA-binding protein-43 proteinopathy in oligodendrocytes revealed using an induced pluripotent stem cell model

Brain Commun. 2021 Oct 26;3(4):fcab255. doi: 10.1093/braincomms/fcab255. eCollection 2021.

Authors

Samantha K Barton^{1

2

3

4}, Dario Magnani^{1

2

3}, Owen G James^{1

2

3}, Matthew R Livesey^{1

5

6}, Bhuvaneish T Selvaraj^{1

2

3}, Owain T James^{1

2

5}, Emma M Perkins^{1

2

5}, Jenna M Gregory^{1

2}, Elaine Cleary^{1

2}, C Rosanne M Ausems^{1

2}, Roderick N Carter Carter^{1

2

3

7}, Navneet A Vasistha^{1

2}, Chen Zhao^{1

2}, Karen Burr^{1

2

3}, David Story^{1

2

3}, Alessandra Cardinali^{1

2

3}, Nicholas M Morton⁷, Giles E Hardingham^{1

3

5}, David J A Wyllie^{1

5

8}, Siddharthan Chandran^{1

2

3

8}

Affiliations

¹ Euan MacDonald Centre for MND, University of Edinburgh, Edinburgh EH16 4SB, UK.
² Centre for Clinical Brain Sciences, University of Edinburgh, Edinburgh EH16 4SB, UK.
³ UK Dementia Research Institute at University of Edinburgh, University of Edinburgh, Edinburgh, EH16 4SB, UK.
⁴ Florey Institute of Neuroscience and Mental Health, Melbourne 3052, Australia.
⁵ Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh EH8 9XD, UK.
⁶ Department of Neuroscience, SITraN, University of Sheffield, Sheffield S10 2HQ, UK.
⁷ Centre for Cardiovascular Science, University of Edinburgh, Edinburgh EH16 4SB, UK.
⁸ Centre for Brain Development and Repair, Institute for Stem Cell Biology and Regenerative Medicine, Bangalore 560065, India.

Abstract

Oligodendrocytes are implicated in amyotrophic lateral sclerosis pathogenesis and display transactive response DNA-binding protein-43 (TDP-43) pathological inclusions. To investigate the cell autonomous consequences of TDP-43 mutations on human oligodendrocytes, we generated oligodendrocytes from patient-derived induced pluripotent stem cell lines harbouring mutations in the TARDBP gene, namely G298S and M337V. Through a combination of immunocytochemistry, electrophysiological assessment via whole-cell patch clamping, and three-dimensional cultures, no differences in oligodendrocyte differentiation, maturation or myelination were identified. Furthermore, expression analysis for monocarboxylate transporter 1 (a lactate transporter) coupled with a glycolytic stress test showed no deficit in lactate export. However, using confocal microscopy, we report TDP-43 mutation-dependent pathological mis-accumulation of TDP-43. Furthermore, using in vitro patch-clamp recordings, we identified functional Ca²⁺-permeable α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor dysregulation in oligodendrocytes. Together, these findings establish a platform for further interrogation of the role of oligodendrocytes and cellular autonomy in TDP-43 proteinopathy.

Keywords: TDP-43; amyotrophic lateral sclerosis; induced pluripotent stem cell; oligodendrocytes.