Abstract

The cytoplasmic mislocalization and aggregation of TAR DNA-binding protein-43 (TDP-43) is a common histopathological hallmark of the amyotrophic lateral sclerosis and frontotemporal dementia disease spectrum (ALS/FTD). However, the composition of aggregates and their contribution to the disease process remain unknown. Here we used proximity-dependent biotin identification (BioID) to interrogate the interactome of detergent-insoluble TDP-43 aggregates and found them enriched for components of the nuclear pore complex and nucleocytoplasmic transport machinery. Aggregated and disease-linked mutant TDP-43 triggered the sequestration and/or mislocalization of nucleoporins and transport factors, and interfered with nuclear protein import and RNA export in mouse primary cortical neurons, human fibroblasts and induced pluripotent stem cell–derived neurons. Nuclear pore pathology is present in brain tissue in cases of sporadic ALS and those involving genetic mutations in TARDBP and C9orf72. Our data strongly implicate TDP-43-mediated nucleocytoplasmic transport defects as a common disease mechanism in ALS/FTD.

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Acknowledgements

We thank K. R. Moss and K. T. Thomas for help with the preparation of primary cortical neurons, G. J. Bassell for logistical support and M. Castanedes-Casey for expert staining of human brain tissue. For numerous expression plasmids used in this study (Supplementary Table 3), we thank M. Hetzer (The Salk Institute for Biological Studies), J. Ellenberg (EMBL Heidelberg), V. Doye (Institut Jacques Monod, Université Paris Diderot/CNRS), J. Teodoro (McGill University) and J. Joseph (National Centre for Cell Science, S.P. Pune University), L. Gerace (The Scripps Research Institute), B. Paschal (University of Virginia School of Medicine) and M. Dasso (Eunice Kennedy Shriver National Institute of Child Health and Human Development). We thank the Bloomington Drosophila Stock Center for fly lines and Emory Integrated Proteomics Core, Neuropathology/ Histochemistry Core and Robert P. Apkarian Integrated Electron Microscopy Core for technical support. This work was supported by grants from the ALS Association (17-IIP-353) to W.R. and (16-IIP-278) to R.S.; the Emory Medicine Catalyst Funding Program to W.R.; Muscular Dystrophy Association (MDA348086) to R.S.; NIH grants K08-NS087121 to C.M.H., P30-NS055077 to the Neuropathology/Histochemistry core of the Emory NINDS Neurosciences Core Facility, AG025688 to Emory’s Alzheimer’s Disease Research Center, NIH R01-NS091299 to D.C.Z., R35-NS097261 to R.R., R01-NS085207 to R.S., R01NS091749 to W.R. and R01-NS093362 to W.R. and T.K., who is also supported by The Bluefield Project to Cure FTD; the Alzheimer’s Drug Discovery Foundation to N.J.C; and NIH R01-AG053960 to N.T.S., who is also supported in part by the Alzheimer’s Association (ALZ), Alzheimer’s Research UK (ARUK), The Michael J. Fox Foundation for Parkinson’s Research (MJFF) and a Weston Brain Institute Biomarkers Across Neurodegenerative Diseases Grant (11060). S.V. was partially funded by UBRP with funds from the UA Provost’s Office. P.G.D.-A. was funded by an ARCS Fellowship Roche Foundation Award.

Author information

Author notes

    • Ching-Chieh Chou

    Present address: Department of Biology, Stanford University, Stanford, CA, USA

    • Yi Zhang

    Present address: Department of Respiratory Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China

    • Paul G. Donlin-Asp

    Present address: Max Planck Institute for Brain Research, Frankfurt, Germany

  1. Ching-Chieh Chou and Yi Zhang contributed equally to this work.

Affiliations

  1. Department of Cell Biology, Emory University School of Medicine, Atlanta, GA, USA

    • Ching-Chieh Chou
    • , Yi Zhang
    • , Paul G. Donlin-Asp
    • , Yu Han Chen
    • , Maureen A. Powers
    •  & Wilfried Rossoll
  2. Center for Neurodegenerative Disease, Emory University School of Medicine, Atlanta, GA, USA

    • Ching-Chieh Chou
    • , Mfon E. Umoh
    • , Duc M. Duong
    • , Nicholas T. Seyfried
    • , Thomas Kukar
    • , Chadwick M. Hales
    • , Marla Gearing
    • , Jonathan D. Glass
    •  & Wilfried Rossoll
  3. Xiangya Hospital and Xiangya School of Medicine, Central South University, Changsha, Hunan, China

    • Yi Zhang
  4. Department of Neurology, Emory University School of Medicine, Atlanta, GA, USA

    • Mfon E. Umoh
    • , Nicholas T. Seyfried
    • , Thomas Kukar
    • , Chadwick M. Hales
    • , Marla Gearing
    •  & Jonathan D. Glass
  5. Department of Molecular & Cellular Biology, University of Arizona, Tucson, AZ, USA

    • Spencer W. Vaughan
    • , Melissa Sayegh
    •  & Daniela C. Zarnescu
  6. Department of Neurobiology, Barrow Neurological Institute, Phoenix, AZ, USA

    • Ileana Lorenzini
    •  & Rita Sattler
  7. Department of Neuroscience, Mayo Clinic, Jacksonville, FL, USA

    • Feilin Liu
    • , Dennis W. Dickson
    • , Rosa Rademakers
    • , Yong-Jie Zhang
    • , Leonard Petrucelli
    •  & Wilfried Rossoll
  8. Department of Ophthalmology, the Second Hospital of Jilin University, Changchun, China

    • Feilin Liu
  9. Department of Biochemistry, Emory University School of Medicine, Atlanta, GA, USA

    • Duc M. Duong
    •  & Nicholas T. Seyfried
  10. Department of Pharmacology, Emory University School of Medicine, Atlanta, GA, USA

    • Thomas Kukar
  11. Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, USA

    • Marla Gearing
  12. Department of Pathology and Immunology, Washington University, St. Louis, MO, USA

    • Nigel J. Cairns
  13. Department of Neurology, Mayo Clinic, Jacksonville, FL, USA

    • Kevin B. Boylan
  14. Emory ALS Center, Emory University School of Medicine, Atlanta, GA, USA

    • Jonathan D. Glass

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Contributions

W.R. conceived and directed the project. C.-C.C., Y.Z. and W.R. designed the experiments. C.-C.C. and W.R. interpreted data and wrote the manuscript. C.-C.C. characterized protein interactome, performed bioinformatic analysis and conducted experiments in N2a cells, primary cortical neurons and human fibroblasts with help from F.L. and Y.H.C. Y.Z. performed the BioID pulldown and sample preparation for LC-MS/MS analysis. M.E.U. performed immunohistochemistry staining. S.W.V., M.S. and D.C.Z. performed Drosophila experiments. I.L. and R.S. performed experiments in iPSC-derived motor neurons. C.-C.C. and P.G.D.-A. conducted SIM experiments. D.M.D., N.T.S. and M.A.P. provided technical support. T.K. provided key reagents. C.M.H., M.G., N.J.C., K.B.B., D.W.D., R.R., Y.-J.Z., L.P. and J.D.G. provided patient tissue with associated clinical and genetics data.

Competing interests

The authors declare that they have no competing financial interests. 

Corresponding author

Correspondence to Wilfried Rossoll.

Supplementary information

  1. Supplementary Text and Figures

    Supplementary Figures 1–17 and Supplementary Tables 1–4

  2. Life Sciences Reporting Summary

  3. Supplementary Video 1: GFP-expressing cells show normal NM morphology

    Three-dimensional (3D) reconstruction of nuclear membrane (NM) staining with anti-lamin B antibody (red) in primary neurons expressing GFP (green).

  4. Supplementary Video 2: GFP-CTF-expressing cells show abnormal NM morphology with deep invaginations

    Three-dimensional (3D) reconstruction of nuclear membrane (NM) staining with anti-lamin B antibody (red) in primary neurons expressing GFP-TDP-CTF (green).

  5. Supplementary Data

    Proteomics list

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https://doi.org/10.1038/s41593-017-0047-3