Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy

Molecular Neurodegeneration

Research Article

James Shorter¹  Ching-Chieh Chou²  Christopher J. Donnelly³  Amanda M. Gleixner³  Björn Oskarsson⁴  Deepak Chhangani⁵  Todd E. Golde⁶  Diego E. Rincon-Limas⁷  Xingli Li⁸  Sami J. Barmada⁸  Cara L. Croft⁹  Bilal Khalil^1,10  Dmytro Morderer^1,10  Feilin Liu^1,10  Ke Zhang^1,10  Dennis W. Dickson^1,10  Christian Puttinger^1,10  Gael Fortin^1,10  Chih-Wei Tsai^1,10  Junli Gao^1,10  Leonard Petrucelli^1,10  Melissa C. Wren^1,10  Wilfried Rossoll^1,10  Jingjiao Chen^1,11  Jannifer H. Lee^1,12  Courtney L. Smith^1,12  Shige H. Yoshimura^1,13  Chun Kim Lim^1,13 

[1] Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, 19104, Philadelphia, PA, USA;Department of Biology, Stanford University, 94305, Stanford, CA, USA;Department of Neurobiology, University of Pittsburgh School of Medicine, 15213, Pittsburgh, PA, USA;LiveLikeLou Center for ALS Research, University of Pittsburgh Brain Institute, 15261, Pittsburgh, PA, USA;Department of Neurology, Mayo Clinic, Jacksonville, FL, USA;Department of Neurology, McKnight Brain Institute, Norman Fixel Institute for Neurological Diseases, University of Florida, 32610, Gainesville, FL, USA;Department of Neurology, McKnight Brain Institute, Norman Fixel Institute for Neurological Diseases, University of Florida, 32610, Gainesville, FL, USA;Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of Florida, 32610, Gainesville, FL, USA;Department of Neurology, McKnight Brain Institute, Norman Fixel Institute for Neurological Diseases, University of Florida, 32610, Gainesville, FL, USA;Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of Florida, 32610, Gainesville, FL, USA;Genetics Institute, University of Florida, 32610, Gainesville, FL, USA;Department of Neurology, University of Michigan, 48109, Ann Arbor, MI, USA;Department of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of Florida, 32610, Gainesville, FL, USA;UK Dementia Research Institute at University College London, London, UK;Department of Neuroscience, Mayo Clinic, 32224, Jacksonville, FL, USA;Department of Neuroscience, Mayo Clinic, 32224, Jacksonville, FL, USA;Geriatric Department, Affiliated Hospital of Qingdao University, Qingdao, Shandong, China;Department of Neuroscience, Mayo Clinic, 32224, Jacksonville, FL, USA;Mayo Clinic Graduate School of Biomedical Sciences, Neuroscience Track, Mayo Clinic, Jacksonville, FL, USA;Graduate School of Biostudies, Kyoto University, Yoshida-konoe, Sakyo-ku, Kyoto, Japan;

关键词: Amyotrophic lateral sclerosis;    Frontotemporal dementia;    TDP-43;    Nucleocytoplasmic transport;    Importin;    Nuclear pore;    Aggregation;    Prion-like domain;    Drosophila;   

DOI  :  10.1186/s13024-022-00585-1

 received in 2022-11-09, accepted in 2022-11-23,  发布年份 2022

来源: Springer

【 摘 要 】

BackgroundCytoplasmic 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.MethodsWe 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.ResultsWe 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.ConclusionOur 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

【 授权许可】

CC BY   
© The Author(s) 2022

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