Inhibition of GCK-IV kinases dissociates cell death and axon regeneration in CNS neurons

Axon injury is a hallmark of many neurodegenerative diseases, often resulting in neuronal cell death and functional impairment. Dual leucine zipper kinase (DLK) has emerged as a key mediator of this process. However, while DLK inhibition is robustly protective in a wide range of neurodegenerative di...

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Published in:Proceedings of the National Academy of Sciences - PNAS 2020-12, Vol.117 (52), p.33597-33607
Main Authors: Patel, Amit K., Broyer, Risa M., Lee, Cassidy D., Lu, Tianlun, Louie, Mikaela J., Torre, Anna La, Al-Ali, Hassan, Vu, Mai T., Mitchell, Katherine L., Wahlin, Karl J., Berlinicke, Cynthia A., Jaskula-Ranga, Vinod, Hu, Yang, Duan, Xin, Vilar, Santiago, Bixby, John L., Weinreb, Robert N., Lemmon, Vance P., Zack, Donald J., Welsbie, Derek S.
Format: Article
Language:English
Subjects:
Animals
Axons - enzymology
Axons - pathology
Base Sequence
Biological Sciences
Cell Death - drug effects
Cell Survival - drug effects
Central Nervous System - pathology
CRISPR-Cas Systems - genetics
Dependovirus - metabolism
Disease Models, Animal
Germinal Center Kinases - metabolism
Humans
Mice, Inbred C57BL
Nerve Regeneration - drug effects
Neuronal Outgrowth - drug effects
Optic Nerve Injuries - metabolism
Optic Nerve Injuries - pathology
Organoids - metabolism
Protein Kinase Inhibitors - pharmacology
Retinal Ganglion Cells - drug effects
Retinal Ganglion Cells - metabolism
Signal Transduction - drug effects
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Summary:Axon injury is a hallmark of many neurodegenerative diseases, often resulting in neuronal cell death and functional impairment. Dual leucine zipper kinase (DLK) has emerged as a key mediator of this process. However, while DLK inhibition is robustly protective in a wide range of neurodegenerative disease models, it also inhibits axonal regeneration. Indeed, there are no genetic perturbations that are known to both improve long-term survival and promote regeneration. To identify such a neuroprotective target, we conducted a set of complementary high-throughput screens using a protein kinase inhibitor library in human stem cell-derived retinal ganglion cells (hRGCs). Overlapping compounds that promoted both neuroprotection and neurite outgrowth were bioinformatically deconvoluted to identify specific kinases that regulated neuronal death and axon regeneration. This work identified the role of germinal cell kinase four (GCK-IV) kinases in cell death and additionally revealed their unexpected activity in suppressing axon regeneration. Using an adeno-associated virus (AAV) approach, coupled with genome editing, we validated that GCK-IV kinase knockout improves neuronal survival, comparable to that of DLK knockout, while simultaneously promoting axon regeneration. Finally, we also found that GCK-IV kinase inhibition also prevented the attrition of RGCs in developing retinal organoid cultures without compromising axon outgrowth, addressing a major issue in the field of stem cell-derived retinas. Together, these results demonstrate a role for the GCK-IV kinases in dissociating the cell death and axonal outgrowth in neurons and their druggability provides for therapeutic options for neurodegenerative diseases.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.2004683117