CRISPR/Cas9 screen in human iPSC-derived cortical neurons identifies NEK6 as a novel disease modifier of C9orf72 poly(PR) toxicity

Wenting Guo, Haibo Wang, Arun Kumar Tharkeshwar, Julien Couthouis, Elke Braems, Pegah Masrori, Evelien Van Schoor, Yannan Fan, Karan Ahuja, Matthieu Moisse, Maarten Jacquemyn, Rodrigo Furtado Madeiro da Costa, Madhavsai Gajjar, Sriram Balusu, Tine Tricot, Laura Fumagalli, Nicole Hersmus, Rekin's Janky, Francis Impens, Pieter Vanden BergheRitchie Ho, Dietmar Rudolf Thal, Rik Vandenberghe, Muralidhar L. Hegde, Siddharthan Chandran, Bart De Strooper, Dirk Daelemans, Philip Van Damme, Ludo Van Den Bosch, Catherine Verfaillie

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Introduction: The most common genetic cause of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) are hexanucleotide repeats in chromosome 9 open reading frame 72 (C9orf72). These repeats produce dipeptide repeat proteins with poly(PR) being the most toxic one. Methods: We performed a kinome-wide CRISPR/Cas9 knock-out screen in human induced pluripotent stem cell (iPSC) -derived cortical neurons to identify modifiers of poly(PR) toxicity, and validated the role of candidate modifiers using in vitro, in vivo, and ex-vivo studies. Results: Knock-down of NIMA-related kinase 6 (NEK6) prevented neuronal toxicity caused by poly(PR). Knock-down of nek6 also ameliorated the poly(PR)-induced axonopathy in zebrafish and NEK6 was aberrantly expressed in C9orf72 patients. Suppression of NEK6 expression and NEK6 activity inhibition rescued axonal transport defects in cortical neurons from C9orf72 patient iPSCs, at least partially by reversing p53-related DNA damage. Discussion: We identified NEK6, which regulates poly(PR)-mediated p53-related DNA damage, as a novel therapeutic target for C9orf72 FTD/ALS.

Original languageEnglish (US)
Pages (from-to)1245-1259
Number of pages15
JournalAlzheimer's and Dementia
Issue number4
StatePublished - Apr 2023


  • C9orf72
  • CRISPR/Cas9 screen
  • DNA damage
  • NEK6
  • PR toxicity
  • amyotrophic lateral sclerosis
  • frontotemporal dementia
  • human pluripotent stem cells
  • neurodegeneration
  • p53
  • DNA Repeat Expansion/genetics
  • C9orf72 Protein/genetics
  • Humans
  • Tumor Suppressor Protein p53/genetics
  • Amyotrophic Lateral Sclerosis/genetics
  • Zebrafish/genetics
  • Frontotemporal Dementia/genetics
  • NIMA-Related Kinases/genetics
  • Animals
  • Neurons/metabolism
  • CRISPR-Cas Systems
  • Induced Pluripotent Stem Cells/metabolism

ASJC Scopus subject areas

  • Clinical Neurology
  • Geriatrics and Gerontology
  • Psychiatry and Mental health
  • Cellular and Molecular Neuroscience
  • Health Policy
  • Developmental Neuroscience
  • Epidemiology


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