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

Tania D Gendron, Kevin F. Bieniek, Yongjie 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 V Rademakers, Kevin B. Boylan, Dennis W Dickson, Leonard Petrucelli

Research output: Contribution to journalArticle

237 Citations (Scopus)

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 cooccur 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.

Original languageEnglish (US)
Pages (from-to)829-844
Number of pages16
JournalActa Neuropathologica
Volume126
Issue number6
DOIs
StatePublished - 2013

Fingerprint

Nuclear RNA
Amyotrophic Lateral Sclerosis
RNA
Neurodegenerative Diseases
Frontal Lobe
Cerebellum
Cultured Cells
Spinal Cord
Proteins
Antibodies
Brain

Keywords

  • Amyotrophic lateral sclerosis
  • Bidirectional transcription
  • C9ORF72
  • Expanded repeat
  • Frontotemporal dementia
  • Repeat-associated non-ATG translation
  • RNA foci

ASJC Scopus subject areas

  • Clinical Neurology
  • Pathology and Forensic Medicine
  • Cellular and Molecular Neuroscience

Cite this

Antisense transcripts of the expanded C9ORF72 hexanucleotide repeat form nuclear RNA foci and undergo repeat-associated non-ATG translation in c9FTD/ALS. / Gendron, Tania D; Bieniek, Kevin F.; Zhang, Yongjie; Jansen-West, Karen; Ash, Peter E A; Caulfield, Thomas; Daughrity, Lillian; Dunmore, Judith H.; Castanedes-Casey, Monica; Chew, Jeannie; Cosio, Danielle M.; Van Blitterswijk, Marka; Lee, Wing C.; Rademakers, Rosa V; Boylan, Kevin B.; Dickson, Dennis W; Petrucelli, Leonard.

In: Acta Neuropathologica, Vol. 126, No. 6, 2013, p. 829-844.

Research output: Contribution to journalArticle

Gendron, Tania D ; Bieniek, Kevin F. ; Zhang, Yongjie ; Jansen-West, Karen ; Ash, Peter E A ; Caulfield, Thomas ; Daughrity, Lillian ; Dunmore, Judith H. ; Castanedes-Casey, Monica ; Chew, Jeannie ; Cosio, Danielle M. ; Van Blitterswijk, Marka ; Lee, Wing C. ; Rademakers, Rosa V ; Boylan, Kevin B. ; Dickson, Dennis W ; Petrucelli, Leonard. / Antisense transcripts of the expanded C9ORF72 hexanucleotide repeat form nuclear RNA foci and undergo repeat-associated non-ATG translation in c9FTD/ALS. In: Acta Neuropathologica. 2013 ; Vol. 126, No. 6. pp. 829-844.
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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 cooccur 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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AU - Zhang, Yongjie

AU - Jansen-West, Karen

AU - Ash, Peter E A

AU - Caulfield, Thomas

AU - Daughrity, Lillian

AU - Dunmore, Judith H.

AU - Castanedes-Casey, Monica

AU - Chew, Jeannie

AU - Cosio, Danielle M.

AU - Van Blitterswijk, Marka

AU - Lee, Wing C.

AU - Rademakers, Rosa V

AU - Boylan, Kevin B.

AU - Dickson, Dennis W

AU - Petrucelli, Leonard

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N2 - 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 cooccur 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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