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

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

  • Alzheimers Dement. 2023 Apr;19(4):1245-1259. doi: 10.1002/alz.12760.
Wenting Guo 1 2 3,  Haibo Wang 4 5,  Arun Kumar Tharkeshwar 2 3,  Julien Couthouis 6,  Elke Braems 2 3,  Pegah Masrori 2 3 7,  Evelien Van Schoor 2 3 8,  Yannan Fan 1,  Karan Ahuja 1,  Matthieu Moisse 2 3,  Maarten Jacquemyn 9,  Rodrigo Furtado Madeiro da Costa 1,  Madhavsai Gajjar 1,  Sriram Balusu 3,  Tine Tricot 1,  Laura Fumagalli 2 3,  Nicole Hersmus 2 3,  Rekin's Janky 10,  Francis Impens 11 12 13,  Pieter Vanden Berghe 14,  Ritchie Ho 15,  Dietmar Rudolf Thal 8,  Rik Vandenberghe 7 16,  Muralidhar L Hegde 4 5,  Siddharthan Chandran 17 18,  Bart De Strooper 3 17,  Dirk Daelemans 9,  Philip Van Damme 2 3 7,  Ludo Van Den Bosch 2 3,  Catherine Verfaillie 1
Affiliations

Affiliations

  • 1 Stem Cell Institute, Department of Devolpment and Regeneration, KU Leuven, Leuven, Belgium.
  • 2 Department of Neurosciences, Experimental Neurology, Laboratory of Neurobiology, KU Leuven-University of Leuven, Leuven, Belgium.
  • 3 VIB, Center for Brain & Disease Research, Leuven, Belgium and Leuven Brain Institute (LBI), Leuven, Belgium.
  • 4 Division of DNA Repair Research, Department of Neurosurgery, Center for Neuroregeneration, Houston Methodist Research Institute, Houston, Texas, USA.
  • 5 Department of Neuroscience Research at Neurological Surgery, Weill Medical College, New York, New York, USA.
  • 6 Department of Genetics, Stanford University School of Medicine, Stanford, California, USA.
  • 7 Department of Neurology, University Hospitals Leuven, Leuven, Belgium.
  • 8 Laboratory of Neuropathology, Department of Imaging and Pathology, KU Leuven, and Leuven Brain Institute (LBI), Leuven, Belgium.
  • 9 KU Leuven Department of Microbiology, Immunology and Transplantation, Laboratory of Virology and Chemotherapy, Rega Institute for Medical Research, Leuven, Belgium.
  • 10 VIB Nucleomics Core, Leuven, Belgium.
  • 11 VIB-UGent Center for Medical Biotechnology, Ghent, Belgium.
  • 12 Department of Biomolecular Medicine, Ghent University, Ghent, Belgium.
  • 13 VIB Proteomics Core, Ghent, Belgium.
  • 14 Translational Research Centre for Gastrointestinal Disorders (TARGID), KU Leuven-University of Leuven, Leuven, Belgium.
  • 15 Center for Neural Science and Medicine, Board of Governors Regenerative Medicine Institute, Departments of Biomedical Sciences and Neurology, Cedars-Sinai Medical Center, Los Angeles, California, USA.
  • 16 KU Leuven-Laboratory for Cognitive Neurology, Department of Neurosciences, Leuven Brain Institute, Leuven, Belgium.
  • 17 UK-Dementia Research Institute at University College London, London, UK.
  • 18 Centre for Clinical Brain Sciences, University of Edinburgh, Edinburgh, UK.
PMID: 35993441 DOI: 10.1002/alz.12760
Abstract

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.

Keywords

C9orf72; CRISPR/Cas9 screen; DNA damage; NEK6; PR toxicity; amyotrophic lateral sclerosis; frontotemporal dementia; human pluripotent stem cells; neurodegeneration; p53.

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