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Open Access 01-12-2013 | Original Paper

Antisense transcripts of the expanded C9ORF72 hexanucleotide repeat form nuclear RNA foci and undergo repeat-associated non-ATG translation in c9FTD/ALS

Authors: Tania F. Gendron, Kevin F. Bieniek, Yong-Jie Zhang, Karen Jansen-West, Peter E. A. Ash, Thomas Caulfield, Lillian Daughrity, Judith H. Dunmore, Monica Castanedes-Casey, Jeannie Chew, Danielle M. Cosio, Marka van Blitterswijk, Wing C. Lee, Rosa Rademakers, Kevin B. Boylan, Dennis W. Dickson, Leonard Petrucelli

Published in: Acta Neuropathologica | Issue 6/2013

Abstract

Frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) are devastating neurodegenerative disorders with clinical, genetic, and neuropathological overlap. A hexanucleotide (GGGGCC) repeat expansion in a non-coding region of C9ORF72 is the major genetic cause of both diseases. The mechanisms by which this repeat expansion causes “c9FTD/ALS” are not definitively known, but RNA-mediated toxicity is a likely culprit. RNA transcripts of the expanded GGGGCC repeat form nuclear foci in c9FTD/ALS, and also undergo repeat-associated non-ATG (RAN) translation resulting in the production of three aggregation-prone proteins. The goal of this study was to examine whether antisense transcripts resulting from bidirectional transcription of the expanded repeat behave in a similar manner. We show that ectopic expression of (CCCCGG)₆₆ in cultured cells results in foci formation. Using novel polyclonal antibodies for the detection of possible (CCCCGG)_exp RAN proteins [poly(PR), poly(GP) and poly(PA)], we validated that (CCCCGG)₆₆ is also subject to RAN translation in transfected cells. Of importance, foci composed of antisense transcripts are observed in the frontal cortex, spinal cord and cerebellum of c9FTD/ALS cases, and neuronal inclusions of poly(PR), poly(GP) and poly(PA) are present in various brain tissues in c9FTD/ALS, but not in other neurodegenerative diseases, including CAG repeat disorders. Of note, RNA foci and poly(GP) inclusions infrequently co-occur in the same cell, suggesting these events represent two distinct ways in which the C9ORF72 repeat expansion may evoke neurotoxic effects. These findings provide mechanistic insight into the pathogenesis of c9FTD/ALS, and have significant implications for therapeutic strategies.

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Title: Antisense transcripts of the expanded C9ORF72 hexanucleotide repeat form nuclear RNA foci and undergo repeat-associated non-ATG translation in c9FTD/ALS
Authors: Tania F. Gendron
Kevin F. Bieniek
Yong-Jie Zhang
Karen Jansen-West
Peter E. A. Ash
Thomas Caulfield
Lillian Daughrity
Judith H. Dunmore
Monica Castanedes-Casey
Jeannie Chew
Danielle M. Cosio
Marka van Blitterswijk
Wing C. Lee
Rosa Rademakers
Kevin B. Boylan
Dennis W. Dickson
Leonard Petrucelli
Publication date: 01-12-2013
Publisher: Springer Berlin Heidelberg
Published in: Acta Neuropathologica / Issue 6/2013
Print ISSN: 0001-6322
Electronic ISSN: 1432-0533
DOI: https://doi.org/10.1007/s00401-013-1192-8

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Antisense transcripts of the expanded C9ORF72 hexanucleotide repeat form nuclear RNA foci and undergo repeat-associated non-ATG translation in c9FTD/ALS

Abstract

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Abstract

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