Targeting RNA foci in iPSC-derived motor neurons from ALS patients with a C9ORF72 repeat expansion

Dhruv Sareen, Jacqueline G. O'Rourke, Pratap Meera, A. K M G Muhammad, Sharday Grant, Megan Simpkinson, Shaughn Bell, Sharon Carmona, Loren Ornelas, Anais Sahabian, Tania D Gendron, Leonard Petrucelli, Michael Baughn, John Ravits, Matthew B. Harms, Frank Rigo, C. Frank Bennett, Thomas S. Otis, Clive N. Svendsen, Robert H. Baloh

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Abstract

Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative condition characterized by loss of motor neurons in the brain and spinal cord. Expansions of a hexanucleotide repeat (GGGGCC) in the noncoding region of the C9ORF72 gene are the most common cause of the familial form of ALS (C9-ALS), as well as frontotemporal lobar degeneration and other neurological diseases. How the repeat expansion causes disease remains unclear, with both loss of function (haploinsufficiency) and gain of function (either toxic RNA or protein products) proposed. We report a cellular model of C9-ALS with motor neurons differentiated from induced pluripotent stem cells (iPSCs) derived from ALS patients carrying the C9ORF72 repeat expansion. No significant loss of C9ORF72 expression was observed, and knockdown of the transcript was not toxic to cultured human motor neurons. Transcription of the repeat was increased, leading to accumulation of GGGGCC repeat-containing RNA foci selectively in C9-ALS iPSC-derived motor neurons. Repeat-containing RNA foci colocalized with hnRNPA1 and Pur-α, suggesting that theymay be able to alter RNA metabolism. C9-ALS motor neurons showed altered expression of genes involved in membrane excitability including DPP6, and demonstrated a diminished capacity to fire continuous spikes upon depolarization compared to control motor neurons. Antisense oligonucleotides targeting the C9ORF72 transcript suppressed RNA foci formation and reversed gene expression alterations in C9-ALS motor neurons. These data show that patient-derived motor neurons can be used to delineate pathogenic events in ALS.

Original languageEnglish (US)
Article number208ra149
JournalScience Translational Medicine
Volume5
Issue number208
DOIs
StatePublished - Oct 23 2013

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Induced Pluripotent Stem Cells
Amyotrophic Lateral Sclerosis
Motor Neurons
RNA
Poisons
Frontotemporal Lobar Degeneration
Gene Expression
Haploinsufficiency
Antisense Oligonucleotides
Spinal Cord
Membranes
Brain

ASJC Scopus subject areas

  • Medicine(all)

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Sareen, D., O'Rourke, J. G., Meera, P., Muhammad, A. K. M. G., Grant, S., Simpkinson, M., ... Baloh, R. H. (2013). Targeting RNA foci in iPSC-derived motor neurons from ALS patients with a C9ORF72 repeat expansion. Science Translational Medicine, 5(208), [208ra149]. https://doi.org/10.1126/scitranslmed.3007529

Targeting RNA foci in iPSC-derived motor neurons from ALS patients with a C9ORF72 repeat expansion. / Sareen, Dhruv; O'Rourke, Jacqueline G.; Meera, Pratap; Muhammad, A. K M G; Grant, Sharday; Simpkinson, Megan; Bell, Shaughn; Carmona, Sharon; Ornelas, Loren; Sahabian, Anais; Gendron, Tania D; Petrucelli, Leonard; Baughn, Michael; Ravits, John; Harms, Matthew B.; Rigo, Frank; Frank Bennett, C.; Otis, Thomas S.; Svendsen, Clive N.; Baloh, Robert H.

In: Science Translational Medicine, Vol. 5, No. 208, 208ra149, 23.10.2013.

Research output: Contribution to journalArticle

Sareen, D, O'Rourke, JG, Meera, P, Muhammad, AKMG, Grant, S, Simpkinson, M, Bell, S, Carmona, S, Ornelas, L, Sahabian, A, Gendron, TD, Petrucelli, L, Baughn, M, Ravits, J, Harms, MB, Rigo, F, Frank Bennett, C, Otis, TS, Svendsen, CN & Baloh, RH 2013, 'Targeting RNA foci in iPSC-derived motor neurons from ALS patients with a C9ORF72 repeat expansion', Science Translational Medicine, vol. 5, no. 208, 208ra149. https://doi.org/10.1126/scitranslmed.3007529
Sareen, Dhruv ; O'Rourke, Jacqueline G. ; Meera, Pratap ; Muhammad, A. K M G ; Grant, Sharday ; Simpkinson, Megan ; Bell, Shaughn ; Carmona, Sharon ; Ornelas, Loren ; Sahabian, Anais ; Gendron, Tania D ; Petrucelli, Leonard ; Baughn, Michael ; Ravits, John ; Harms, Matthew B. ; Rigo, Frank ; Frank Bennett, C. ; Otis, Thomas S. ; Svendsen, Clive N. ; Baloh, Robert H. / Targeting RNA foci in iPSC-derived motor neurons from ALS patients with a C9ORF72 repeat expansion. In: Science Translational Medicine. 2013 ; Vol. 5, No. 208.
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abstract = "Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative condition characterized by loss of motor neurons in the brain and spinal cord. Expansions of a hexanucleotide repeat (GGGGCC) in the noncoding region of the C9ORF72 gene are the most common cause of the familial form of ALS (C9-ALS), as well as frontotemporal lobar degeneration and other neurological diseases. How the repeat expansion causes disease remains unclear, with both loss of function (haploinsufficiency) and gain of function (either toxic RNA or protein products) proposed. We report a cellular model of C9-ALS with motor neurons differentiated from induced pluripotent stem cells (iPSCs) derived from ALS patients carrying the C9ORF72 repeat expansion. No significant loss of C9ORF72 expression was observed, and knockdown of the transcript was not toxic to cultured human motor neurons. Transcription of the repeat was increased, leading to accumulation of GGGGCC repeat-containing RNA foci selectively in C9-ALS iPSC-derived motor neurons. Repeat-containing RNA foci colocalized with hnRNPA1 and Pur-α, suggesting that theymay be able to alter RNA metabolism. C9-ALS motor neurons showed altered expression of genes involved in membrane excitability including DPP6, and demonstrated a diminished capacity to fire continuous spikes upon depolarization compared to control motor neurons. Antisense oligonucleotides targeting the C9ORF72 transcript suppressed RNA foci formation and reversed gene expression alterations in C9-ALS motor neurons. These data show that patient-derived motor neurons can be used to delineate pathogenic events in ALS.",
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AU - Baughn, Michael

AU - Ravits, John

AU - Harms, Matthew B.

AU - Rigo, Frank

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AU - Baloh, Robert H.

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N2 - Amyotrophic lateral sclerosis (ALS) is a severe neurodegenerative condition characterized by loss of motor neurons in the brain and spinal cord. Expansions of a hexanucleotide repeat (GGGGCC) in the noncoding region of the C9ORF72 gene are the most common cause of the familial form of ALS (C9-ALS), as well as frontotemporal lobar degeneration and other neurological diseases. How the repeat expansion causes disease remains unclear, with both loss of function (haploinsufficiency) and gain of function (either toxic RNA or protein products) proposed. We report a cellular model of C9-ALS with motor neurons differentiated from induced pluripotent stem cells (iPSCs) derived from ALS patients carrying the C9ORF72 repeat expansion. No significant loss of C9ORF72 expression was observed, and knockdown of the transcript was not toxic to cultured human motor neurons. Transcription of the repeat was increased, leading to accumulation of GGGGCC repeat-containing RNA foci selectively in C9-ALS iPSC-derived motor neurons. Repeat-containing RNA foci colocalized with hnRNPA1 and Pur-α, suggesting that theymay be able to alter RNA metabolism. C9-ALS motor neurons showed altered expression of genes involved in membrane excitability including DPP6, and demonstrated a diminished capacity to fire continuous spikes upon depolarization compared to control motor neurons. Antisense oligonucleotides targeting the C9ORF72 transcript suppressed RNA foci formation and reversed gene expression alterations in C9-ALS motor neurons. These data show that patient-derived motor neurons can be used to delineate pathogenic events in ALS.

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