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CRISPR-Cas9 genetic screen leads to the discovery of L-Moses, a KAT2B inhibitor that attenuates Tunicamycin-mediated neuronal cell death.

Published version
Peer-reviewed

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Authors

Pavlou, Sofia 
Foskolou, Stefanie 
Patikas, Nikolaos 
Field, Sarah F 
Papachristou, Evangelia K 

Abstract

Accumulation of aggregated and misfolded proteins, leading to endoplasmic reticulum stress and activation of the unfolded protein response, is a hallmark of several neurodegenerative disorders, including Alzheimer's and Parkinson's disease. Genetic screens are powerful tools that are proving invaluable in identifying novel modulators of disease associated processes. Here, we performed a loss-of-function genetic screen using a human druggable genome library, followed by an arrayed-screen validation, in human iPSC-derived cortical neurons. We identified and genetically validated 13 genes, whose knockout was neuroprotective against Tunicamycin, a glycoprotein synthesis inhibitor widely used to induce endoplasmic reticulum stress. We also demonstrated that pharmacological inhibition of KAT2B, a lysine acetyltransferase identified by our genetic screens, by L-Moses, attenuates Tunicamycin-mediated neuronal cell death and activation of CHOP, a key pro-apoptotic member of the unfolded protein response in both cortical and dopaminergic neurons. Follow-up transcriptional analysis suggested that L-Moses provided neuroprotection by partly reversing the transcriptional changes caused by Tunicamycin. Finally, L-Moses treatment attenuated total protein levels affected by Tunicamycin, without affecting their acetylation profile. In summary, using an unbiased approach, we identified KAT2B and its inhibitor, L-Moses, as potential therapeutic targets for neurodegenerative diseases.

Description

Acknowledgements: We would like to thank Dr. Andrew Bassett for his invaluable help throughout the project, Dr. Mark Kotter for kindly gifting the iPSCs NGN2-OPTi-OX cell line and Dr Michael E. Ward for gifting the NGN2-iPSCs. This work was supported by Open Targets (grant OTAR2054). EM was funded by UK Dementia Research Institute (UK DRI) grant RRZA/175. Diagrams were created using BioRender (biorender.com). GO analysis was perform using Metascape (metascape.org).

Keywords

Humans, Tunicamycin, CRISPR-Cas Systems, Endoplasmic Reticulum, Cell Death, Endoplasmic Reticulum Stress, Dopaminergic Neurons, Apoptosis, p300-CBP Transcription Factors

Journal Title

Sci Rep

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Journal ISSN

2045-2322
2045-2322

Volume Title

Publisher

Springer Science and Business Media LLC
Sponsorship
UK Dementia Research Institute (DRICam17/18)