Antisense RNA foci in the motor neurons of C9ORF72-ALS patients are associated with TDP-43 proteinopathy

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Web End = Acta Neuropathol (2015) 130:6375 DOI 10.1007/s00401-015-1429-9

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Web End = Antisense RNA foci in the motor neurons of C9ORF72ALS patients are associated with TDP43 proteinopathy

Johnathan CooperKnock1 Adrian Higginbottom1 Matthew J. Stopford1 J. Robin Highley1 Paul G. Ince1 Stephen B. Wharton1 Stuart PickeringBrown2 Janine Kirby1 Guillaume M. Hautbergue1 Pamela J. Shaw1

Abstract GGGGCC repeat expansions of C9ORF72 represent the most common genetic variant of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia. We and others have proposed that RNA transcribed from the repeat sequence is toxic via sequestration of RNA-binding factors. Both GGGGCC-repeat (sense) and CCCCGG-repeat (antisense) molecules are detectable by uorescence in situ hybridisation as RNA foci, but their relative expression pattern within the CNS and contribution to disease has not been determined. Blinded examination of CNS biosamples from ALS patients with a repeat expansion of C9ORF72 showed that antisense foci are present at a signicantly higher frequency in cerebellar Purkinje neurons and motor neurons, whereas sense foci are present at a signicantly higher frequency in cerebellar granule neurons. Consistent with this, inclusions containing sense or antisense derived dipeptide repeat proteins were present at signicantly higher frequency in cerebellar granule neurons or motor neurons, respectively. Immunohistochemistry and UV-crosslinking studies showed that sense and antisense RNA molecules share similar interactions with SRSF2, hnRNP K, hnRNP A1, ALYREF, and hnRNP H/F. Together these

Received: 19 February 2015 / Revised: 8 April 2015 / Accepted: 20 April 2015 / Published online: 6 May 2015 The Author(s) 2015. This article is published with open access at Springerlink.com

data suggest that, although sense and antisense RNA molecules might be expected to be equally toxic via their shared protein binding partners, distinct patterns of expression in various CNS neuronal populations could lead to relative differences in their contribution to the pathogenesis of neuronal injury. Moreover in motor neurons, which are the primary target of pathology in ALS, the presence of antisense foci (2, p < 0.00001) but not sense foci (2, p = 0.75)

correlated with mislocalisation of TDP-43, which is the hallmark of ALS neurodegeneration. This...