Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosis

Cytoplasmic mislocalisation and nuclear depletion of TDP-43 are pathological hallmarks of amyotrophic lateral sclerosis (ALS), including mutations in the C9ORF72 gene that characterise the most common genetic form of ALS (C9ALS). Studies in human cells and animal models have associated cytoplasmic m...

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Main Authors: Manpreet Singh Atwal, Jerneja Nimac, Urša Čerček, Sarah Ricarda Goesch, Hannah Rebecca Goesch, Paraskevi Tziortzouda, Tiziana Ercolani, Anna Zatorska, Terouz Pasha, Ivo Carre, Jacqueline Mitchell, Claire Troakes, Bart Tummers, Vera Župunski, Boris Rogelj, Tibor Hortobágyi, Frank Hirth
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Neuroscience
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Online Access:https://www.frontiersin.org/articles/10.3389/fnins.2025.1558227/full
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author Manpreet Singh Atwal
Jerneja Nimac
Jerneja Nimac
Urša Čerček
Urša Čerček
Sarah Ricarda Goesch
Hannah Rebecca Goesch
Paraskevi Tziortzouda
Tiziana Ercolani
Anna Zatorska
Terouz Pasha
Ivo Carre
Jacqueline Mitchell
Claire Troakes
Bart Tummers
Vera Župunski
Boris Rogelj
Boris Rogelj
Tibor Hortobágyi
Tibor Hortobágyi
Frank Hirth
author_facet Manpreet Singh Atwal
Jerneja Nimac
Jerneja Nimac
Urša Čerček
Urša Čerček
Sarah Ricarda Goesch
Hannah Rebecca Goesch
Paraskevi Tziortzouda
Tiziana Ercolani
Anna Zatorska
Terouz Pasha
Ivo Carre
Jacqueline Mitchell
Claire Troakes
Bart Tummers
Vera Župunski
Boris Rogelj
Boris Rogelj
Tibor Hortobágyi
Tibor Hortobágyi
Frank Hirth
author_sort Manpreet Singh Atwal
collection DOAJ
description Cytoplasmic mislocalisation and nuclear depletion of TDP-43 are pathological hallmarks of amyotrophic lateral sclerosis (ALS), including mutations in the C9ORF72 gene that characterise the most common genetic form of ALS (C9ALS). Studies in human cells and animal models have associated cytoplasmic mislocalisation of TDP-43 with abnormalities in nuclear transport receptors, referred to as karyopherins, that mediate the nucleocytoplasmic shuttling of TDP-43. Yet the relationship between karyopherin abnormalities and TDP-43 pathology are unclear. Here we report karyopherin-α4 (KPNA4) pathology in the spinal cord of TDP-43-positive sporadic ALS and C9ALS patients. Structural analyses revealed the selective interaction between KPNA subtypes, especially KPNA4, with the nuclear localisation signal (NLS) of TDP-43. Targeted cytoplasmic mislocalisation and nuclear depletion of TDP-43 caused KPNA4 pathology in human cells. Similar phenotypes were observed in Drosophila whereby cytoplasmic accumulation of the TDP-43 homolog, TBPH, caused the nuclear decrease and cytosolic mislocalisation of the KPNA4 homolog, Importin-α3 (Impα3). In contrast, induced accumulation of Impα3 was not sufficient to cause TBPH mislocalisation. Instead, targeted gain of Impα3 in the presence of accumulating cytosolic TBPH, restored Impα3 localisation and partially rescued nuclear TBPH. These results demonstrate that cytoplasmic accumulation of TDP-43 causes karyopherin pathology that characterises ALS spinal cord. Together with earlier reports, our findings establish KPNA4 abnormalities as a molecular signature of TDP-43 proteinopathies and identify it as a potential therapeutic target to sustain nuclear TDP-43 essential for cellular homeostasis affected in ALS and frontotemporal dementia.
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spelling doaj-art-54d0aaf01b124b4ab409cb1c1ce367312025-08-20T03:25:27ZengFrontiers Media S.A.Frontiers in Neuroscience1662-453X2025-07-011910.3389/fnins.2025.15582271558227Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosisManpreet Singh Atwal0Jerneja Nimac1Jerneja Nimac2Urša Čerček3Urša Čerček4Sarah Ricarda Goesch5Hannah Rebecca Goesch6Paraskevi Tziortzouda7Tiziana Ercolani8Anna Zatorska9Terouz Pasha10Ivo Carre11Jacqueline Mitchell12Claire Troakes13Bart Tummers14Vera Župunski15Boris Rogelj16Boris Rogelj17Tibor Hortobágyi18Tibor Hortobágyi19Frank Hirth20Department of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Biotechnology, Jozef Stefan Institute, Ljubljana, SloveniaGraduate School of Biomedicine, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaDepartment of Biotechnology, Jozef Stefan Institute, Ljubljana, SloveniaGraduate School of Biomedicine, Faculty of Medicine, University of Ljubljana, Ljubljana, SloveniaDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomLondon Neurodegenerative Diseases Brain Bank, Department of Basic and Clinical Neuroscience, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomCentre for Inflammation Biology and Cancer Immunology, King’s College London, London, United KingdomFaculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, SloveniaDepartment of Biotechnology, Jozef Stefan Institute, Ljubljana, SloveniaFaculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, SloveniaDepartment of Neurology, University of Debrecen, Debrecen, HungaryCentre for Healthy Brain Ageing, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomDepartment of Basic and Clinical Neuroscience, Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry, Psychology and Neuroscience, King's College London, London, United KingdomCytoplasmic mislocalisation and nuclear depletion of TDP-43 are pathological hallmarks of amyotrophic lateral sclerosis (ALS), including mutations in the C9ORF72 gene that characterise the most common genetic form of ALS (C9ALS). Studies in human cells and animal models have associated cytoplasmic mislocalisation of TDP-43 with abnormalities in nuclear transport receptors, referred to as karyopherins, that mediate the nucleocytoplasmic shuttling of TDP-43. Yet the relationship between karyopherin abnormalities and TDP-43 pathology are unclear. Here we report karyopherin-α4 (KPNA4) pathology in the spinal cord of TDP-43-positive sporadic ALS and C9ALS patients. Structural analyses revealed the selective interaction between KPNA subtypes, especially KPNA4, with the nuclear localisation signal (NLS) of TDP-43. Targeted cytoplasmic mislocalisation and nuclear depletion of TDP-43 caused KPNA4 pathology in human cells. Similar phenotypes were observed in Drosophila whereby cytoplasmic accumulation of the TDP-43 homolog, TBPH, caused the nuclear decrease and cytosolic mislocalisation of the KPNA4 homolog, Importin-α3 (Impα3). In contrast, induced accumulation of Impα3 was not sufficient to cause TBPH mislocalisation. Instead, targeted gain of Impα3 in the presence of accumulating cytosolic TBPH, restored Impα3 localisation and partially rescued nuclear TBPH. These results demonstrate that cytoplasmic accumulation of TDP-43 causes karyopherin pathology that characterises ALS spinal cord. Together with earlier reports, our findings establish KPNA4 abnormalities as a molecular signature of TDP-43 proteinopathies and identify it as a potential therapeutic target to sustain nuclear TDP-43 essential for cellular homeostasis affected in ALS and frontotemporal dementia.https://www.frontiersin.org/articles/10.3389/fnins.2025.1558227/fullamyotrophic lateral sclerosisTDP-43C9ORF72karyopherinKPNA4nuclear import
spellingShingle Manpreet Singh Atwal
Jerneja Nimac
Jerneja Nimac
Urša Čerček
Urša Čerček
Sarah Ricarda Goesch
Hannah Rebecca Goesch
Paraskevi Tziortzouda
Tiziana Ercolani
Anna Zatorska
Terouz Pasha
Ivo Carre
Jacqueline Mitchell
Claire Troakes
Bart Tummers
Vera Župunski
Boris Rogelj
Boris Rogelj
Tibor Hortobágyi
Tibor Hortobágyi
Frank Hirth
Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosis
Frontiers in Neuroscience
amyotrophic lateral sclerosis
TDP-43
C9ORF72
karyopherin
KPNA4
nuclear import
title Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosis
title_full Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosis
title_fullStr Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosis
title_full_unstemmed Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosis
title_short Accumulation of TDP-43 causes karyopherin-α4 pathology that characterises amyotrophic lateral sclerosis
title_sort accumulation of tdp 43 causes karyopherin α4 pathology that characterises amyotrophic lateral sclerosis
topic amyotrophic lateral sclerosis
TDP-43
C9ORF72
karyopherin
KPNA4
nuclear import
url https://www.frontiersin.org/articles/10.3389/fnins.2025.1558227/full
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