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Autophagy induction enhances TDP43 turnover and survival in neuronal ALS models

Nature Chemical Biology volume 10, pages 677–685 (2014)Cite this article

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Abstract

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) have distinct clinical features but a common pathology—cytoplasmic inclusions rich in transactive response element DNA-binding protein of 43 kDa (TDP43). Rare TDP43 mutations cause ALS or FTD, but abnormal TDP43 levels and localization may cause disease even if TDP43 lacks a mutation. Here we show that individual neurons vary in their ability to clear TDP43 and are exquisitely sensitive to TDP43 levels. To measure TDP43 clearance, we developed and validated a single-cell optical method that overcomes the confounding effects of aggregation and toxicity and discovered that pathogenic mutations shorten TDP43 half-life. New compounds that stimulate autophagy improved TDP43 clearance and localization and enhanced survival in primary murine neurons and in human stem cell–derived neurons and astrocytes harboring mutant TDP43. These findings indicate that the levels and localization of TDP43 critically determine neurotoxicity and show that autophagy induction mitigates neurodegeneration by acting directly on TDP43 clearance.

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Figure 1: The toxicity of TDP43 depends strongly on dose.
Figure 2: Optical pulse-labeling of TDP43 in primary neurons.
Figure 3: OPL of TDP43.
Figure 4: Induction of autophagy by a family of small molecules.
Figure 5: Autophagy induction reduces TDP43 levels, aggregation and cytoplasmic mislocalization.
Figure 6: Autophagic stimulation improves survival in neuronal and astrocyte models of ALS.

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Acknowledgements

We would like to thank B. Miller, B. Seeley, C. Lomen-Hoerth and the members of the Finkbeiner lab for their generous support and advice. H. Zahed and J. Margulis deserve special acknowledgment for their assistance. We thank G. Howard for editorial assistance and K. Nelson for administrative assistance. We also thank G. Yu and J. Herz (University of Texas, Southwestern) for anti-TDP43 antibodies. This work was supported by National Institutes of Neurological Disorders and Stroke grants K08 NS072233 (to S.J.B.), 3R01 NS039074, R01 NS083390, R43 NS081844 and U24 NS078370 (to S.F.); the ALS Association (S.F.); the Robert Packard Center for ALS Research and the William H. Adams Foundation (S.F.); and Target ALS (S.F.). Additional support was provided by the Roddenberry Stem Cell Program (to S.F.), the Taube-Koret Center for Neurodegenerative Disease (S.F.), the Hellman Family Foundation Alzheimer's Disease Research Program (S.F.), the Protein Folding Diseases Initiative at the University of Michigan (S.J.B.) and the California Institute of Regenerative Medicine TR4-06693 (S.F.) and U01 MH1050135 (S.F.). The animal care facility was partly supported by an US National Institutes of Health Extramural Research Facilities Improvement Program Project (C06 RR018928).

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Authors and Affiliations

  1. Gladstone Institute of Neurologic Disease, San Francisco, California, USA

    Sami J Barmada, Arpana Arjun, Aaron Daub, D Michael Ando, Andrey Tsvetkov & Steven Finkbeiner

  2. Department of Neurology, University of California–San Francisco Medical Center, San Francisco, California, USA

    Sami J Barmada & Steven Finkbeiner

  3. Department of Neurology, University of Michigan, Ann Arbor, Michigan, USA

    Sami J Barmada, Xingli Li & Daniel Peisach

  4. Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, Scotland, UK

    Andrea Serio, Bilada Bilican & Siddharthan Chandran

  5. Medical Research Council Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, Scotland, UK

    Andrea Serio, Bilada Bilican & Siddharthan Chandran

  6. Departments of Materials and Bioengineering, and Institute for Biomedical Engineering, Imperial College, London, UK

    Andrea Serio

  7. Biomedical Sciences Graduate Program, University of California–San Francisco, San Francisco, California, USA

    D Michael Ando & Steven Finkbeiner

  8. Department of Neurobiology and Anatomy, University of Texas Medical School, Houston, Texas, USA

    Andrey Tsvetkov

  9. Keck Program in Brain Cell Engineering, Gladstone Institutes, San Francisco, California, USA

    Michael Pleiss & Steven Finkbeiner

  10. Institute of Psychiatry, King's College London, London, UK

    Christopher Shaw

  11. Department of Physiology, University of California–San Francisco, San Francisco, California, USA

    Steven Finkbeiner

  12. Taube-Koret Center for Neurodegenerative Disease Research, San Francisco, California, USA

    Steven Finkbeiner

  13. Hellman Family Foundation Alzheimer's Disease Research Program, San Francisco, California, USA

    Steven Finkbeiner

  14. Roddenberry Stem Cell Program, San Francisco, California, USA

    Steven Finkbeiner

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Contributions

S.J.B., A.S., A.A., A.T., S.C. and S.F. designed the study. S.J.B., A.A., X.L. and A.S. cultured cells, performed microscopy and conducted survival analyses. A.T., B.B., C.S. and S.C. provided compounds and iPSCs. M.P. performed in silico drug screening. D.P., D.M.A. and A.D. wrote original scripts for AFM. S.J.B. and A.A. analyzed data, constructed vectors, performed MPC and OPL and transfected neurons. S.B. and S.F. wrote and edited the manuscript.

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Correspondence to Steven Finkbeiner.

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Barmada, S., Serio, A., Arjun, A. et al. Autophagy induction enhances TDP43 turnover and survival in neuronal ALS models. Nat Chem Biol 10, 677–685 (2014). https://doi.org/10.1038/nchembio.1563

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