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Quantitative analysis and clinico-pathological correlations of different dipeptide repeat protein pathologies in C9ORF72 mutation carriers

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Abstract

Hexanucleotide repeat expansion in C9ORF72 is the most common genetic cause of frontotemporal dementia and motor neuron disease. One consequence of the mutation is the formation of different potentially toxic polypeptides composed of dipeptide repeats (DPR) (poly-GA, -GP, -GR, -PA, -PR) generated by repeat-associated non-ATG (RAN) translation. While previous studies focusing on poly-GA pathology have failed to detect any clinico-pathological correlations in C9ORF72 mutation cases, recent data from animal and cell culture models suggested that it may be only specific DPR species that are toxic and only when accumulated in certain intracellular compartments. Therefore, we performed a systematic clinico-pathological correlative analysis with counting of actual numbers of distinct types of inclusion (neuronal cytoplasmic and intranuclear inclusions, dystrophic neurites) for each DPR protein in relevant brain regions (premotor cortex, lower motor neurons) in a cohort of 35 C9ORF72 mutation cases covering the clinical spectrum from those with pure MND, mixed FTD/MND and pure FTD. While each DPR protein pathology had a similar pattern of anatomical distribution, the total amount of inclusions for each DPR protein varied remarkably (poly-GA > GP > GR > PR/PA), indicating that RAN translation seems to be more effective from sense than from antisense transcripts. Importantly, with the exception of moderate associations for the amount of poly-GA-positive dystrophic neurites with degeneration in the frontal cortex and total burden of poly-GA pathology with disease onset, no relationship was identified for any other DPR protein pathology with degeneration or phenotype. Biochemical analysis revealed a close correlation between insoluble DPR protein species and numbers of visible inclusions, while we did not find any evidence for the presence of soluble DPR protein species. Thus, overall our findings strongly argue against a role of DPR protein aggregation as major and exclusive pathomechanism in C9ORF72 pathogenesis. However, this does not exclude that DPR protein formation might be essential in C9ORF72 pathogenesis in interplay with other consequences associated with the C9ORF72 repeat expansion.

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Acknowledgments

We would like to thank Margaret Luk, Katrin Trautmann and Manuel Gödan for their excellent technical assistance. This work was supported by the German Helmholtz Association (VH-VI-510 and W2/W3-036, MN), the Canadian Institutes of Health Research (74580, IRM), the Pacific Alzheimer’s Research Foundation (C06-01, IRM), and the European Unions’s Seventh Framework Program (FP7/2014-2019 Grant 617198, DE).

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Authors and Affiliations

  1. Department of Pathology, University of British Columbia and Vancouver General Hospital, Vancouver, Canada

    Ian R. A. Mackenzie

  2. German Center for Neurodegenerative Diseases (DZNE), Otfried-Müllerstr. 23, 72076, Tübingen, Germany

    Petra Frick & Manuela Neumann

  3. Institute of Virology, Saarland University Medical School, Homburg, Germany

    Friedrich A. Grässer

  4. Department of Neuroscience, Mayo Clinic Florida, Jacksonville, USA

    Tania F. Gendron & Leonard Petrucelli

  5. Department of Medicine (Neurology), Brain Research Centre, University of British Columbia, Vancouver, Canada

    Neil R. Cashman

  6. German Center for Neurodegenerative Diseases (DZNE), Munich, Germany

    Dieter Edbauer

  7. Munich Cluster of Systems Neurology (SyNergy), Munich, Germany

    Dieter Edbauer & Elisabeth Kremmer

  8. Institute for Metabolic Biochemistry, Ludwig-Maximilians University Munich, Munich, Germany

    Dieter Edbauer

  9. Institute of Molecular Immunology, Helmholtz Zentrum München, Munich, Germany

    Elisabeth Kremmer

  10. German Center for Neurodegenerative Diseases (DZNE), Rostock, Germany

    Johannes Prudlo

  11. Department of Neurology, University of Rostock, Rostock, Germany

    Johannes Prudlo

  12. Department of Neuropathology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands

    Dirk Troost

  13. Department of Neuropathology, University of Tübingen, Tübingen, Germany

    Manuela Neumann

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  1. Ian R. A. Mackenzie
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Correspondence to Manuela Neumann.

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Mackenzie, I.R.A., Frick, P., Grässer, F.A. et al. Quantitative analysis and clinico-pathological correlations of different dipeptide repeat protein pathologies in C9ORF72 mutation carriers. Acta Neuropathol 130, 845–861 (2015). https://doi.org/10.1007/s00401-015-1476-2

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