eprintid: 10167363 rev_number: 6 eprint_status: archive userid: 699 dir: disk0/10/16/73/63 datestamp: 2023-03-28 09:44:20 lastmod: 2023-03-28 09:44:20 status_changed: 2023-03-28 09:44:20 type: article metadata_visibility: show sword_depositor: 699 creators_name: Khalil, Bilal creators_name: Chhangani, Deepak creators_name: Wren, Melissa C creators_name: Smith, Courtney L creators_name: Lee, Jannifer H creators_name: Li, Xingli creators_name: Puttinger, Christian creators_name: Tsai, Chih-Wei creators_name: Fortin, Gael creators_name: Morderer, Dmytro creators_name: Gao, Junli creators_name: Liu, Feilin creators_name: Lim, Chun Kim creators_name: Chen, Jingjiao creators_name: Chou, Ching-Chieh creators_name: Croft, Cara L creators_name: Gleixner, Amanda M creators_name: Donnelly, Christopher J creators_name: Golde, Todd E creators_name: Petrucelli, Leonard creators_name: Oskarsson, Bjorn creators_name: Dickson, Dennis W creators_name: Zhang, Ke creators_name: Shorter, James creators_name: Yoshimura, Shige H creators_name: Barmada, Sami J creators_name: Rincon-Limas, Diego E creators_name: Rossoll, Wilfried title: Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy ispublished: pub divisions: UCL divisions: B02 divisions: C07 divisions: D07 divisions: FD9 keywords: Science & Technology, Life Sciences & Biomedicine, Neurosciences, Neurosciences & Neurology, Amyotrophic lateral sclerosis, Frontotemporal dementia, TDP-43, Nucleocytoplasmic transport, Importin, Nuclear pore, Aggregation, Prion-like domain, Drosophila, FRONTOTEMPORAL LOBAR DEGENERATION, PHASE-SEPARATION, NUCLEOCYTOPLASMIC TRANSPORT, PTDP-43 PATHOLOGY, BINDING, ALS, FUS, BETA, LOCALIZATION, TRANSITIONS note: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. 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The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. abstract: Background: Cytoplasmic mislocalization and aggregation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of the amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD) disease spectrum, causing both nuclear loss-of-function and cytoplasmic toxic gain-of-function phenotypes. While TDP-43 proteinopathy has been associated with defects in nucleocytoplasmic transport, this process is still poorly understood. Here we study the role of karyopherin-β1 (KPNB1) and other nuclear import receptors in regulating TDP-43 pathology. Methods: We used immunostaining, immunoprecipitation, biochemical and toxicity assays in cell lines, primary neuron and organotypic mouse brain slice cultures, to determine the impact of KPNB1 on the solubility, localization, and toxicity of pathological TDP-43 constructs. Postmortem patient brain and spinal cord tissue was stained to assess KPNB1 colocalization with TDP-43 inclusions. Turbidity assays were employed to study the dissolution and prevention of aggregation of recombinant TDP-43 fibrils in vitro. Fly models of TDP-43 proteinopathy were used to determine the effect of KPNB1 on their neurodegenerative phenotype in vivo. Results: We discovered that several members of the nuclear import receptor protein family can reduce the formation of pathological TDP-43 aggregates. Using KPNB1 as a model, we found that its activity depends on the prion-like C-terminal region of TDP-43, which mediates the co-aggregation with phenylalanine and glycine-rich nucleoporins (FG-Nups) such as Nup62. KPNB1 is recruited into these co-aggregates where it acts as a molecular chaperone that reverses aberrant phase transition of Nup62 and TDP-43. These findings are supported by the discovery that Nup62 and KPNB1 are also sequestered into pathological TDP-43 aggregates in ALS/FTD postmortem CNS tissue, and by the identification of the fly ortholog of KPNB1 as a strong protective modifier in Drosophila models of TDP-43 proteinopathy. Our results show that KPNB1 can rescue all hallmarks of TDP-43 pathology, by restoring its solubility and nuclear localization, and reducing neurodegeneration in cellular and animal models of ALS/FTD. Conclusion: Our findings suggest a novel NLS-independent mechanism where, analogous to its canonical role in dissolving the diffusion barrier formed by FG-Nups in the nuclear pore, KPNB1 is recruited into TDP-43/FG-Nup co-aggregates present in TDP-43 proteinopathies and therapeutically reverses their deleterious phase transition and mislocalization, mitigating neurodegeneration. Graphical Abstract: [Figure not available: see fulltext.]. date: 2022-12-08 date_type: published publisher: BMC official_url: https://doi.org/10.1186/s13024-022-00585-1 oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1994651 doi: 10.1186/s13024-022-00585-1 medium: Electronic pii: 10.1186/s13024-022-00585-1 lyricists_name: Croft, Cara lyricists_id: CCROF38 actors_name: Flynn, Bernadette actors_id: BFFLY94 actors_role: owner funding_acknowledgements: [Robert Packard Center for ALS Research]; [Mayo Clinic Center for Biomedical Discovery]; A2021038S [BrightFocus foundation]; R33NS110960 [National Institutes of Health (NIH)]; RF1AG068581 [National Institutes of Health (NIH)]; R01AG077771 [National Institutes of Health (NIH)]; P01NS084974 [National Institutes of Health (NIH)]; P30AG062677 [National Institutes of Health (NIH)]; R01NS097542 [National Institutes of Health (NIH)]; R01NS113943 [National Institutes of Health (NIH)]; P30AG072931 [National Institutes of Health (NIH)]; R01NS117461 [National Institutes of Health (NIH)]; R01NS105756 [National Institutes of Health (NIH)]; R21AG064940 [National Institutes of Health (NIH)]; R01NS127187 [National Institutes of Health (NIH)]; R35NS097273 [National Institutes of Health (NIH)]; U54NS123743 [National Institutes of Health (NIH)]; R01AG059871 [National Institutes of Health (NIH)]; R21NS096647 [National Institutes of Health (NIH)]; W81XWH-19-1-0193 [US Department of Defense]; [Mayo Clinic Ventures Innovation Loan]; P01NS084974 [Audrey Lewis Young Investigator Award from the CureSMA foundation]; P30AG062677 [Audrey Lewis Young Investigator Award from the CureSMA foundation]; [Rainwater Charitable Foundation]; [Robert E. Jacoby Professorship]; [LiveLikeLou Center at the University of Pittsburgh Brain Institute]; [Robert Packard Center for ALS Research at Johns Hopkins]; [Target ALS Foundation] full_text_status: public publication: Molecular Neurodegeneration volume: 17 article_number: 80 pages: 27 event_location: England issn: 1750-1326 citation: Khalil, Bilal; Chhangani, Deepak; Wren, Melissa C; Smith, Courtney L; Lee, Jannifer H; Li, Xingli; Puttinger, Christian; ... Rossoll, Wilfried; + view all <#> Khalil, Bilal; Chhangani, Deepak; Wren, Melissa C; Smith, Courtney L; Lee, Jannifer H; Li, Xingli; Puttinger, Christian; Tsai, Chih-Wei; Fortin, Gael; Morderer, Dmytro; Gao, Junli; Liu, Feilin; Lim, Chun Kim; Chen, Jingjiao; Chou, Ching-Chieh; Croft, Cara L; Gleixner, Amanda M; Donnelly, Christopher J; Golde, Todd E; Petrucelli, Leonard; Oskarsson, Bjorn; Dickson, Dennis W; Zhang, Ke; Shorter, James; Yoshimura, Shige H; Barmada, Sami J; Rincon-Limas, Diego E; Rossoll, Wilfried; - view fewer <#> (2022) Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy. Molecular Neurodegeneration , 17 , Article 80. 10.1186/s13024-022-00585-1 <https://doi.org/10.1186/s13024-022-00585-1>. Green open access document_url: https://discovery-pp.ucl.ac.uk/id/eprint/10167363/1/Nuclear%20import%20receptors%20are%20recruited%20by%20FG-nucleoporins%20to%20rescue%20hallmarks%20of%20TDP-43%20proteinopathy.pdf