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Author Notes:

Jie Jiang, jie.jiang@emory.edu

Conceptualization, Resources, Formal analysis, Validation, Investigation, Methodology, Writing - original draft, Project administration, Writing - review and editing. Data curation, Validation, Methodology. Formal analysis, Methodology. Formal analysis, Validation. Investigation. Methodology. Methodology. Methodology. Resources. Investigation. Resources. Resources. Resources. Resources, Supervision, Investigation. Resources, Supervision, Funding acquisition, Investigation, Methodology, Writing - original draft, Project administration, Writing - review and editing.

We would like to thank Dr. Jonathan Glass for providing access to the clinical material (Emory cohort) and Dr. Homa Ghalei for thoughtful discussions. We would like to thank Ganesh Chilukuri and Daniel Pun for helping with the neuronal cell death counting. We are grateful to Dr. Yao Yao (University of Georgia) and Dr. Zachary McEachin (Emory University) for providing sense and control ASOs, respectively. JP is supported by the Milton Safenowitz Postdoctoral Fellowship from the ALS association (21-PDF-585). DCP is supported by Milton Safenowitz Postdoctoral Fellowship from the ALS association (22-PDF-605). AB and GJB are supported by the NIH R01 (R01NS114253 to GJB). EB is supported by a PhD Fellowship from FWO-Vlaanderen (1145621N). LVDB is supported by a research project of FWO-Vlaanderen (G0C1620N). The work was supported by the NIH R01 grant (R01AG068247 to JJ) and NIH R21 grant (5R21NS114908-02 to GJB and JJ).

No competing interests declared.

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Research Funding:

This paper was supported by the following grants:

National Institute of Neurological Disorders and Stroke R01NS114253 to Anwesha Banerjee, Gary J Bassell.

National Institutes of Health R01AG068247 to Jie Jiang.

ALS Association 21-PDF-585 to Janani Parameswaran.

Fonds Wetenschappelijk Onderzoek G0C1620N to Ludo Van Den Bosch.

ALS Association 22-PDF-605 to Devesh C Pant.

NIH R01NS114253 to Gary J Bassell.

FWO-Vlaanderen 1145621N to Elke Braems.

FWO-Vlaanderen G0C1620N to Ludo Van Den Bosch.

NIH 5R21NS114908-02 to Gary J Bassell, Jie Jiang.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Biology
  • Life Sciences & Biomedicine - Other Topics
  • C9ORF72
  • PKR
  • antisense RNA
  • FTD
  • ALS
  • integrated stress response
  • Zebrafish
  • FRONTOTEMPORAL LOBAR DEGENERATION
  • AMYOTROPHIC-LATERAL-SCLEROSIS
  • UNCONVENTIONAL TRANSLATION
  • HEXANUCLEOTIDE REPEAT
  • RAN TRANSLATION
  • PROTEIN-KINASE
  • EXPANSION
  • TOXICITY
  • FOCI

Antisense, but not sense, repeat expanded RNAs activate PKR/eIF2a-dependent ISR in C9ORF72 FTD/ALS

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Journal Title:

ELIFE

Volume:

Volume 12

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Type of Work:

Article | Final Publisher PDF

Abstract:

GGGGCC (G4C2) hexanucleotide repeat expansion in the C9ORF72 gene is the most common genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). The repeat is bidirectionally transcribed and confers gain of toxicity. However, the underlying toxic species is debated, and it is not clear whether antisense CCCCGG (C4G2) repeat expanded RNAs contribute to disease pathogenesis. Our study shows that C9ORF72 antisense C4G2 repeat expanded RNAs trigger the activation of the PKR/eIF2α-dependent integrated stress response independent of dipeptide repeat proteins that are produced through repeat-associated non-AUG initiated translation, leading to global translation inhibition and stress granule formation. Reducing PKR levels with either siRNA or morpholinos mitigates integrated stress response and toxicity caused by the antisense C4G2 RNAs in cell lines, primary neurons, and zebrafish. Increased phosphorylation of PKR/eIF2α is also observed in the frontal cortex of C9ORF72 FTD/ALS patients. Finally, only antisense C4G2, but not sense G4C2, repeat expanded RNAs robustly activate the PKR/eIF2α pathway and induce aberrant stress granule formation. These results provide a mechanism by which antisense C4G2 repeat expanded RNAs elicit neuronal toxicity in FTD/ALS caused by C9ORF72 repeat expansions.
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