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Dipeptide repeat protein pathology in C9ORF72 mutation cases: clinico-pathological correlations

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Abstract

Hexanucleotide repeat expansion in C9ORF72 is the most common genetic cause of frontotemporal dementia and motor neuron disease. Recently, unconventional non-ATG translation of the expanded hexanucleotide repeat, resulting in the production and aggregation of dipeptide repeat (DPR) proteins (poly-GA, -GR and GP), was identified as a potential pathomechanism of C9ORF72 mutations. Besides accumulation of DPR proteins, the second neuropathological hallmark lesion in C9ORF72 mutation cases is the accumulation of TDP-43. In this study, we characterized novel monoclonal antibodies against poly-GA and performed a detailed analysis of the neuroanatomical distribution of DPR and TDP-43 pathology in a cohort of 35 cases with the C9ORF72 mutation that included a broad spectrum of clinical phenotypes. We found the pattern of DPR pathology to be highly consistent among cases regardless of the phenotype with high DPR load in the cerebellum, all neocortical regions (frontal, motor cortex and occipital) and hippocampus, moderate pathology in subcortical areas and minimal pathology in lower motor neurons. No correlation between DPR pathology and the degree of neurodegeneration was observed, while a good association between TDP-43 pathology with clinical phenotype and degeneration in key anatomical regions was present. Our data confirm that the presence of DPR pathology is intimately related to C9ORF72 mutations. The observed dissociation between DPR inclusion body load and neurodegeneration might suggest inclusion body formation as a potentially protective response to cope with soluble toxic DPR species. Moreover, our data imply that alterations due to the C9ORF72 mutation resulting in TDP-43 accumulation and dysmetabolism as secondary downstream effects likely play a central role in the neurodegenerative process in C9ORF72 pathogenesis.

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References

  1. Al-Sarraj S, King A, Troakes C et al (2011) p62 positive, TDP-43 negative, neuronal cytoplasmic and intranuclear inclusions in the cerebellum and hippocampus define the pathology of C9orf72-linked FTLD and MND/ALS. Acta Neuropathol 122:691–702. doi:10.1007/s00401-011-0911-2

    Article  PubMed  CAS  Google Scholar 

  2. Arighi A, Fumagalli GG, Jacini F et al (2012) Early onset behavioral variant frontotemporal dementia due to the C9ORF72 hexanucleotide repeat expansion: psychiatric clinical presentations. J Alzheimers Dis 31:447–452. doi:10.3233/JAD-2012-120523

    PubMed  Google Scholar 

  3. Arrasate M, Mitra S, Schweitzer ES, Segal MR, Finkbeiner S (2004) Inclusion body formation reduces levels of mutant huntingtin and the risk of neuronal death. Nature 431:805–810. doi:10.1038/nature02998

    Article  PubMed  CAS  Google Scholar 

  4. Ash PE, Bieniek KF, Gendron TF et al (2013) Unconventional translation of C9ORF72 GGGGCC expansion generates insoluble polypeptides specific to c9FTD/ALS. Neuron. doi:10.1016/j.neuron.2013.02.004

    PubMed  Google Scholar 

  5. Beck J, Poulter M, Hensman D et al (2013) Large C9orf72 hexanucleotide repeat expansions are seen in multiple neurodegenerative syndromes and are more frequent than expected in the UK population. Am J Hum Genet 92:345–353. doi:10.1016/j.ajhg.2013.01.011

    Article  PubMed  CAS  Google Scholar 

  6. Belzil VV, Gendron TF, Petrucelli L (2012) RNA-mediated toxicity in neurodegenerative disease. Mol Cell Neurosci. doi:10.1016/j.mcn.2012.12.006

    PubMed  Google Scholar 

  7. Boeve BF, Boylan KB, Graff-Radford NR et al (2012) Characterization of frontotemporal dementia and/or amyotrophic lateral sclerosis associated with the GGGGCC repeat expansion in C9ORF72. Brain 135:765–783

    Article  PubMed  Google Scholar 

  8. Brettschneider J, Van Deerlin VM, Robinson JL et al (2012) Pattern of ubiquilin pathology in ALS and FTLD indicates presence of C9ORF72 hexanucleotide expansion. Acta Neuropathol 123:825–839. doi:10.1007/s00401-012-0970-z

    Article  PubMed  Google Scholar 

  9. Byrne S, Elamin M, Bede P et al (2012) Cognitive and clinical characteristics of patients with amyotrophic lateral sclerosis carrying a C9orf72 repeat expansion: a population-based cohort study. Lancet Neurol 11:232–240

    Article  PubMed  CAS  Google Scholar 

  10. Cairns NJ, Neumann M, Bigio EH et al (2007) TDP-43 in familial and sporadic frontotemporal lobar degeneration with ubiquitin inclusions. Am J Pathol 171:227–240

    Article  PubMed  CAS  Google Scholar 

  11. Cleary JD, Ranum LP (2013) Repeat-associated non-ATG (RAN) translation in neurological disease. Hum Mol Genet. doi:10.1093/hmg/ddt371

    PubMed  Google Scholar 

  12. Cooper-Knock J, Hewitt C, Highley JR et al (2012) Clinico-pathological features in amyotrophic lateral sclerosis with expansions in C9ORF72. Brain 135:751–764

    Article  PubMed  Google Scholar 

  13. Cruts M, Gijselinck I, Van Langenhove T, van der Zee J, Van Broeckhoven C (2013) Current insights into the C9orf72 repeat expansion diseases of the FTLD/ALS spectrum. Trends Neurosci 36:450–459. doi:10.1016/j.tins.2013.04.010

    Article  PubMed  CAS  Google Scholar 

  14. Dejesus-Hernandez M, Mackenzie IR, Boeve BF et al (2011) Expanded GGGGCC hexanucleotide repeat in noncoding region of C9ORF72 causes chromosome 9p-Linked FTD and ALS. Neuron 72:245–256

    Article  PubMed  CAS  Google Scholar 

  15. Floris G, Borghero G, Cannas A et al (2012) Frontotemporal dementia with psychosis, parkinsonism, visuo-spatial dysfunction, upper motor neuron involvement associated to expansion of C9ORF72: a peculiar phenotype? J Neurol. doi:10.1007/s00415-012-6444-3

    PubMed  Google Scholar 

  16. Galimberti D, Fenoglio C, Serpente M et al (2013) Autosomal dominant frontotemporal lobar degeneration due to the C9ORF72 hexanucleotide repeat expansion: late-onset psychotic clinical presentation. Biol Psychiatry 74:384–391. doi:10.1016/j.biopsych.2013.01.031

    Article  PubMed  CAS  Google Scholar 

  17. Gijselinck I, Van Langenhove T, van der Zee J et al (2012) A C9orf72 promoter repeat expansion in a Flanders–Belgian cohort with disorders of the frontotemporal lobar degeneration–amyotrophic lateral sclerosis spectrum: a gene identification study. Lancet Neurol 11:54–65

    Article  PubMed  CAS  Google Scholar 

  18. Gutekunst CA, Li SH, Yi H et al (1999) Nuclear and neuropil aggregates in Huntington’s disease: relationship to neuropathology. J Neurosci 19:2522–2534

    PubMed  CAS  Google Scholar 

  19. Hsiung GY, Dejesus-Hernandez M, Feldman HH et al (2012) Clinical and pathological features of familial frontotemporal dementia caused by C9ORF72 mutation on chromosome 9p. Brain 135:709–722

    Article  PubMed  Google Scholar 

  20. Lansbury PT, Lashuel HA (2006) A century-old debate on protein aggregation and neurodegeneration enters the clinic. Nature 443:774–779. doi:10.1038/nature05290

    Article  PubMed  CAS  Google Scholar 

  21. Mackenzie IR, Bigio EH, Ince PG et al (2007) Pathological TDP-43 distinguishes sporadic amyotrophic lateral sclerosis from amyotrophic lateral sclerosis with SOD1 mutations. Ann Neurol 61:427–434

    Article  PubMed  CAS  Google Scholar 

  22. Mackenzie IR, Neumann M, Baborie A et al (2011) A harmonized classification system for FTLD-TDP pathology. Acta Neuropathol 122:111–113. doi:10.1007/s00401-011-0845-8

    Article  PubMed  Google Scholar 

  23. Mahoney CJ, Beck J, Rohrer JD et al (2012) Frontotemporal dementia with the C9ORF72 hexanucleotide repeat expansion: clinical, neuroanatomical and neuropathological features. Brain 135:736–750

    Article  PubMed  Google Scholar 

  24. Mori K, Weng SM, Arzberger T et al (2013) The C9orf72 GGGGCC repeat is translated into aggregating dipeptide-repeat proteins in FTLD/ALS. Science 339:1335–1338. doi:10.1126/science.1232927

    Article  PubMed  CAS  Google Scholar 

  25. Moseley ML, Zu T, Ikeda Y et al (2006) Bidirectional expression of CUG and CAG expansion transcripts and intranuclear polyglutamine inclusions in spinocerebellar ataxia type 8. Nat Genet 38:758–769. doi:10.1038/ng1827

    Article  PubMed  CAS  Google Scholar 

  26. Murray ME, Dejesus-Hernandez M, Rutherford NJ et al (2011) Clinical and neuropathologic heterogeneity of c9FTD/ALS associated with hexanucleotide repeat expansion in C9ORF72. Acta Neuropathol 122:673–690. doi:10.1007/s00401-011-0907-y

    Article  PubMed  CAS  Google Scholar 

  27. Nelson PT, Alafuzoff I, Bigio EH et al (2012) Correlation of Alzheimer disease neuropathologic changes with cognitive status: a review of the literature. J Neuropathol Exp Neurol 71:362–381. doi:10.1097/NEN.0b013e31825018f7

    Article  PubMed  Google Scholar 

  28. Neumann M, Kwong LK, Lee EB et al (2009) Phosphorylation of S409/410 of TDP-43 is a consistent feature in all sporadic and familial forms of TDP-43 proteinopathies. Acta Neuropathol 117:137–149

    Article  PubMed  CAS  Google Scholar 

  29. Neumann M, Sampathu DM, Kwong LK et al (2006) Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Science 314:130–133

    Article  PubMed  CAS  Google Scholar 

  30. Orr HT (2013) Toxic RNA as a driver of disease in a common form of ALS and dementia. Proc Natl Acad Sci USA 110:7533–7534. doi:10.1073/pnas.1305239110

    Article  PubMed  CAS  Google Scholar 

  31. Renton AE, Majounie E, Waite A et al (2011) A Hexanucleotide Repeat Expansion in C9ORF72 Is the Cause of Chromosome 9p21-Linked ALS-FTD. Neuron 72:257–268

    Article  PubMed  CAS  Google Scholar 

  32. Schnell SA, Staines WA, Wessendorf MW (1999) Reduction of lipofuscin-like autofluorescence in fluorescently labeled tissue. J Histochem Cytochem Off J Histochem Soc 47:719–730

    Article  CAS  Google Scholar 

  33. Simon-Sanchez J, Dopper EG, Cohn-Hokke PE et al (2012) The clinical and pathological phenotype of C9ORF72 hexanucleotide repeat expansions. Brain 135:723–735

    Article  PubMed  Google Scholar 

  34. Snowden JS, Rollinson S, Thompson JC et al (2012) Distinct clinical and pathological characteristics of frontotemporal dementia associated with C9ORF72 mutations. Brain 135:693–708

    Article  PubMed  Google Scholar 

  35. Stewart H, Rutherford NJ, Briemberg H et al (2012) Clinical and pathological features of amyotrophic lateral sclerosis caused by mutation in the C9ORF72 gene on chromosome 9p. Acta Neuropathol 123:409–417. doi:10.1007/s00401-011-0937-5

    Article  PubMed  CAS  Google Scholar 

  36. Todd PK, Oh SY, Krans A et al (2013) CGG repeat-associated translation mediates neurodegeneration in fragile X tremor ataxia syndrome. Neuron 78:440–455. doi:10.1016/j.neuron.2013.03.026

    Article  PubMed  CAS  Google Scholar 

  37. Treusch S, Cyr DM, Lindquist S (2009) Amyloid deposits: protection against toxic protein species? Cell Cycle 8:1668–1674

    Article  PubMed  CAS  Google Scholar 

  38. Troakes C, Maekawa S, Wijesekera L et al (2011) An MND/ALS phenotype associated with C9orf72 repeat expansion: abundant p62-positive, TDP-43-negative inclusions in cerebral cortex, hippocampus and cerebellum but without associated cognitive decline. Neuropathology. doi:10.1111/j.1440-1789.2011.01286.x

    Google Scholar 

  39. van der Zee J, Gijselinck I, Dillen L et al (2013) A pan-European study of the C9orf72 repeat associated with FTLD: geographic prevalence, genomic instability, and intermediate repeats. Hum Mutat 34:363–373. doi:10.1002/humu.22244

    Article  PubMed  Google Scholar 

  40. Van Langenhove T, van der Zee J, Gijselinck I et al (2013) Distinct clinical characteristics of C9orf72 expansion carriers compared with GRN, MAPT, and nonmutation carriers in a Flanders–Belgian FTLD cohort. JAMA Neurol 70:365–373. doi:10.1001/2013.jamaneurol.181

    Article  PubMed  Google Scholar 

  41. Zu T, Gibbens B, Doty NS et al (2011) Non-ATG-initiated translation directed by microsatellite expansions. Proc Natl Acad Sci USA 108:260–265. doi:10.1073/pnas.1013343108

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We would like to thank Katrin Trautmann, Margaret Luk, Brigitte Kraft and Michael Schmidt for their excellent technical assistance. This work was supported by grants from the German Helmholtz Association (VH-VI-510, MN), the German Federal Ministry of Education and Research (01GI0704, MN), the Alexander von Humboldt Foundation (KM), the Canadian Institutes of Health Research (74580, IRM), the Pacific Alzheimer’s Research Foundation (C06-01, IRM), the Centres of Excellence in Neurodegeneration Research (CoEN, DE and CH) and the Helmholtz Young Investigator (DE) and W2/W3 program (MN).

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Correspondence to Dieter Edbauer or Manuela Neumann.

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I. R. Mackenzie and T. Arzberger contributed equally to the study.

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Below is the link to the electronic supplementary material. DPR inclusions are restricted to neurons. Double-labeling immunofluorescence with cell type specific markers in green, poly-GA in red and merged images with Hoechst nuclei staining in blue. Poly-GA-positive cytoplasmic inclusions are localized to NeuN-positive neurons as shown in the frontal cortex of a C9ORF72 mutation case (a). Poly-GA-positive inclusions are not found in GFAP-positive astrocytes (b), Iba1-positive microglia cells (c) or in CNPase-positive oligodendrocytes (d). Scale bar 25 μm.

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Mackenzie, I.R., Arzberger, T., Kremmer, E. et al. Dipeptide repeat protein pathology in C9ORF72 mutation cases: clinico-pathological correlations. Acta Neuropathol 126, 859–879 (2013). https://doi.org/10.1007/s00401-013-1181-y

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  • DOI: https://doi.org/10.1007/s00401-013-1181-y

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