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 (original) (raw)

Abstract

Neuronal cytoplasmic inclusions (NCIs) containing phosphorylated TDP-43 (p-TDP-43) are the pathological hallmarks of motor neuron disease/amyotrophic lateral sclerosis (MND/ALS) and FTLD-TDP. The vast majority of NCIs in the brain and spinal cord also label for ubiquitin and p62, however, we have previously reported a subset of TDP-43 proteinopathy patients who have unusual and abundant p62 positive, TDP-43 negative inclusions in the cerebellum and hippocampus. Here we sought to determine whether these cases carry the hexanucleotide repeat expansion in C9orf72. Repeat primer PCR was performed in 36 MND/ALS, FTLD-MND/ALS and FTLD-TDP cases and four controls. Fourteen individuals with the repeat expansion were detected. In all the 14 expansion mutation cases there were abundant globular and star-shaped p62 positive NCIs in the pyramidal cell layer of the hippocampus, the vast majority of which were p-TDP-43 negative. p62 positive NCIs were also abundant in the cerebellar granular and molecular layers in all cases and in Purkinje cells in 12/14 cases but they were only positive for p-TDP-43 in the granular layer of one case. Abundant p62 positive, p-TDP-43 negative neuronal intranuclear inclusions (NIIs) were seen in 12/14 cases in the pyramidal cell layer of the hippocampus and in 6/14 cases in the cerebellar granular layer. This unusual combination of inclusions appears pathognomonic for C9orf72 repeat expansion positive MND/ALS and FTLD-TDP which we believe form a pathologically distinct subset of TDP-43 proteinopathies. Our results suggest that proteins other than TDP-43 are binding p62 and aggregating in response to the mutation which may play a mechanistic role in neurodegeneration.

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References

1. Alafuzoff I, Arzberger T, Al-Sarraj S et al (2008) Staging of neurofibrillary pathology in Alzheimer’s disease: a study of the BrainNet Europe Consortium. Brain Pathol 18:484–496
   PubMed Google Scholar
2. Arai T, Hasegawa M, Akiyama H et al (2006) TDP-43 is a component of ubiquitin-positive tau-negative inclusions in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Biochem Biophys Res Commun 351:602–611
   Article PubMed CAS Google Scholar
3. Becher MW, Kotzuk JA, Sharp AH et al (1998) Intranuclear neuronal inclusions in Huntington’s disease and dentatorubral and pallidoluysian atrophy: correlation between the density of inclusions and IT15 CAG triplet repeat length. Neurobiol Dis 4:387–397
   Article PubMed CAS Google Scholar
4. Beck J, Rohrer JD, Campbell T et al (2008) A distinct clinical, neuropsychological and radiological phenotype is associated with progranulin gene mutations in a large UK series. Brain 131:706–720
   Article PubMed Google Scholar
5. Boxer AL, Mackenzie IR, Boeve BF et al (2010) Clinical, neuroimaging and neuropathological features of a new chromosome 9p-linked FTD-ALS family. J Neurol Neurosurg Psychiatry 82:196–203
   Article PubMed Google Scholar
6. Cairns NJ, Bigio EH, Mackenzie IR et al (2007) Neuropathologic diagnostic and nosologic criteria for frontotemporal lobar degeneration: consensus of the consortium for frontotemporal lobar degeneration. Acta Neuropathol 114:5–22
   Article PubMed Google Scholar
7. 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
8. 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
9. Deng HX, Chen W, Hong ST et al (2011) Mutations in UBQLN2 cause dominant X-linked juvenile and adult-onset ALS and ALS/dementia. Nature 477:211–215
   Article PubMed CAS Google Scholar
10. Geser F, Brandmeir NJ, Kwong LK et al (2008) Evidence of multisystem disorder in whole-brain map of pathological TDP-43 in amyotrophic lateral sclerosis. Arch Neurol 65:636–641
    Article PubMed Google Scholar
11. Jackson M, Lennox G, Lowe J (1996) Motor neurone disease-inclusion dementia. Neurodegeneration 5:339–350
    Article PubMed CAS Google Scholar
12. Johnson JO, Mandrioli J, Benatar M et al (2010) Exome sequencing reveals VCP mutations as a cause of familial ALS. Neuron 68:857–864
    Article PubMed CAS Google Scholar
13. King A, Al-Sarraj S, Shaw C (2009) Frontotemporal lobar degeneration with ubiquitinated tau-negative inclusions and additional alpha-synuclein pathology but also unusual cerebellar ubiquitinated p62-positive, TDP-43-negative inclusions. Neuropathology 29:466–471
    Article PubMed Google Scholar
14. King A, Maekawa S, Bodi I et al (2011) Ubiquitinated, p62 immunopositive cerebellar cortical neuronal inclusions are evident across the spectrum of TDP-43 proteinopathies but are only rarely additionally immunopositive for phosphorylation-dependent TDP-43. Neuropathology 31:239–249
    Article PubMed Google Scholar
15. Laaksovirta H, Peuralinna T, Schymick JC et al (2010) Chromosome 9p21 in amyotrophic lateral sclerosis in Finland: a genome-wide association study. Lancet Neurol 9:978–985
    Article PubMed CAS Google Scholar
16. Leigh PN, Anderton BH, Dodson A et al (1988) Ubiquitin deposits in anterior horn cells in motor neurone disease. Neurosci Lett 93:197–203
    Article PubMed CAS Google Scholar
17. Lowe J, Lennox G, Jefferson D et al (1988) A filamentous inclusion body within anterior horn neurones in motor neurone disease defined by immunocytochemical localisation of ubiquitin. Neurosci Lett 94:203–210
    Article PubMed CAS Google Scholar
18. Mackenzie IR (2007) The neuropathology and clinical phenotype of FTD with progranulin mutations. Acta Neuropathol 114:49–54
    Article PubMed Google Scholar
19. Mackenzie IR, Baker M, Pickering-Brown S et al (2006) The neuropathology of frontotemporal lobar degeneration caused by mutations in the progranulin gene. Brain 129:3081–3090
    Article PubMed Google Scholar
20. Mackenzie IR, Neumann M, Bigio EH et al (2010) Nomenclature and nosology for neuropathologic subtypes of frontotemporal lobar degeneration: an update. Acta Neuropathol 119:1–4
    Article PubMed Google Scholar
21. Maekawa S, Leigh PN, King A et al (2009) TDP-43 is consistently co-localized with ubiquitinated inclusions in sporadic and Guam amyotrophic lateral sclerosis but not in familial amyotrophic lateral sclerosis with and without SOD1 mutations. Neuropathology 29:672–683
    Article PubMed Google Scholar
22. Maruyama H, Morino H, Ito H et al (2010) Mutations of optineurin in amyotrophic lateral sclerosis. Nature 465:223–226
    Article PubMed CAS Google Scholar
23. Morita M, Al-Chalabi A, Andersen PM et al (2006) A locus on chromosome 9p confers susceptibility to ALS and frontotemporal dementia. Neurology 66:839–844
    Article PubMed CAS Google Scholar
24. Neumann M, Mackenzie IR, Cairns NJ et al (2007) TDP-43 in the ubiquitin pathology of frontotemporal dementia with VCP gene mutations. J Neuropathol Exp Neurol 66:152–157
    Article PubMed Google Scholar
25. 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
26. Pikkarainen M, Hartikainen P, Alafuzoff I (2008) Neuropathologic features of frontotemporal lobar degeneration with ubiquitin-positive inclusions visualized with ubiquitin-binding protein p62 immunohistochemistry. J Neuropathol Exp Neurol 67:280–298
    Article PubMed Google Scholar
27. 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
28. Sampathu DM, Neumann M, Kwong LK et al (2006) Pathological heterogeneity of frontotemporal lobar degeneration with ubiquitin-positive inclusions delineated by ubiquitin immunohistochemistry and novel monoclonal antibodies. Am J Pathol 169:1343–1352
    Article PubMed CAS Google Scholar
29. Shatunov A, Mok K, Newhouse S et al (2010) Chromosome 9p21 in sporadic amyotrophic lateral sclerosis in the UK and seven other countries: a genome-wide association study. Lancet Neurol 9:986–994
    Article PubMed CAS Google Scholar
30. Sreedharan J, Blair IP, Tripathi VB et al (2008) TDP-43 mutations in familial and sporadic amyotrophic lateral sclerosis. Science 319:1668–1672
    Article PubMed CAS Google Scholar
31. Todd PK, Paulson HL (2010) RNA-mediated neurodegeneration in repeat expansion disorders. Ann Neurol 67:291–300
    Article PubMed CAS Google Scholar
32. Van Deerlin VM, Sleiman PM, Martinez-Lage M et al (2010) Common variants at 7p21 are associated with frontotemporal lobar degeneration with TDP-43 inclusions. Nat Genet 42:234–239
    Article PubMed Google Scholar
33. Vance C, Al-Chalabi A, Ruddy D et al (2006) Familial amyotrophic lateral sclerosis with frontotemporal dementia is linked to a locus on chromosome 9p13.2-21.3. Brain 129:868–876
    Article PubMed Google Scholar
34. Wooten MW, Hu X, Babu JR et al (2006) Signaling, polyubiquitination, trafficking, and inclusions: sequestosome 1/p62’s role in neurodegenerative disease. J Biomed Biotechnol 2006:62079
    Article PubMed Google Scholar

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Acknowledgments

The authors thank the donors and their families whose donation of brain and spinal cord tissue to the Medical Research Council London Neurodegenerative Diseases Brain Bank allowed this research to take place. The authors thank Professor Martin Rossor for referral of GRN mutation cases. They also thank the staff of the Clinical Neuropathology Department, King’s College Hospital, especially Mary Davitt and Joanne Hickey. Funding to support this work came from the Motor Neuron Disease Association, American Amyotrophic Lateral Sclerosis Association, Heaton-Ellis Trust, Medical Research Council (UK), Wellcome Trust and Psychiatry Research Trust.

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The authors declare they have no conflict of interest.

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Authors and Affiliations

1. Department of Clinical Neuropathology, Academic Neuroscience Building, Kings College Hospital, Denmark Hill, London, SE5 9RS, UK
   Safa Al-Sarraj, Andrew King, Satomi Maekawa & Istvan Bodi
2. MRC London Neurodegenerative Diseases Brain Bank, Institute of Psychiatry, Kings College, London, UK
   Safa Al-Sarraj, Andrew King, Claire Troakes, Satomi Maekawa, Istvan Bodi, Ammar Al-Chalabi, Tibor Hortobágyi & Christopher E. Shaw
3. Department of Clinical Neuroscience, MRC Centre for Neurodegeneration Research, Institute of Psychiatry, Kings College, London, UK
   Safa Al-Sarraj, Claire Troakes, Bradley Smith, Satomi Maekawa, Boris Rogelj, Ammar Al-Chalabi, Tibor Hortobágyi & Christopher E. Shaw

Authors

1. Safa Al-Sarraj
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2. Andrew King
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3. Claire Troakes
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4. Bradley Smith
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5. Satomi Maekawa
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6. Istvan Bodi
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7. Boris Rogelj
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8. Ammar Al-Chalabi
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9. Tibor Hortobágyi
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10. Christopher E. Shaw
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Correspondence toAndrew King.

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The authors S. Al-Sarraj and A. King have made an equal contribution.

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Al-Sarraj, S., King, A., Troakes, C. et al. 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 (2011). https://doi.org/10.1007/s00401-011-0911-2

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• Received: 01 November 2011
• Revised: 09 November 2011
• Accepted: 10 November 2011
• Published: 19 November 2011
• Issue Date: December 2011
• DOI: https://doi.org/10.1007/s00401-011-0911-2

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