PUBLICATION

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

Authors
Guo, W., Wang, H., Kumar Tharkeshwar, A., Couthouis, J., Braems, E., Masrori, P., Van Schoor, E., Fan, Y., Ahuja, K., Moisse, M., Jacquemyn, M., Furtado Madeiro da Costa, R., Gajjar, M., Balusu, S., Tricot, T., Fumagalli, L., Hersmus, N., Janky, R., Impens, F., Vanden Berghe, P., Ho, R., Thal, D.R., Vandenberghe, R., Hegde, M.L., Chandran, S., De Strooper, B., Daelemans, D., Van Damme, P., Van Den Bosch, L., Verfaillie, C.
ID
ZDB-PUB-220824-17
Date
2022
Source
Alzheimer's & dementia : the journal of the Alzheimer's Association   19(4): 1245-1259 (Journal)
Registered Authors
Keywords
C9orf72, CRISPR/Cas9 screen, DNA damage, NEK6, PR toxicity, amyotrophic lateral sclerosis, frontotemporal dementia, human pluripotent stem cells, neurodegeneration, p53
MeSH Terms
  • Amyotrophic Lateral Sclerosis*/genetics
  • Animals
  • C9orf72 Protein/genetics
  • CRISPR-Cas Systems
  • DNA Repeat Expansion/genetics
  • Frontotemporal Dementia*/genetics
  • Humans
  • Induced Pluripotent Stem Cells*/metabolism
  • NIMA-Related Kinases/genetics
  • NIMA-Related Kinases/metabolism
  • Neurons/metabolism
  • Tumor Suppressor Protein p53/genetics
  • Tumor Suppressor Protein p53/metabolism
  • Zebrafish/genetics
  • Zebrafish/metabolism
PubMed
35993441 Full text @ Alzheimers Dement
Abstract
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.
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.
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.
We identified NEK6, which regulates poly(PR)-mediated p53-related DNA damage, as a novel therapeutic target for C9orf72 FTD/ALS.
Genes / Markers
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Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping