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Stem Cell Reports. 2019 Mar 5;12(3):502-517. doi: 10.1016/j.stemcr.2019.01.015. Epub 2019 Feb 14.

Dual Inhibition of GSK3β and CDK5 Protects the Cytoskeleton of Neurons from Neuroinflammatory-Mediated Degeneration In Vitro and In Vivo.

Author information

1
Technische Universität Dresden, DFG-Research Center for Regenerative Therapies Dresden (CRTD), Fetscherstrasse 105, 01307 Dresden, Germany; Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149 Münster, Germany.
2
Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149 Münster, Germany; Lead Discovery Center GmbH, Otto-Hahn-Strasse 15, 44227 Dortmund, Germany.
3
Lead Discovery Center GmbH, Otto-Hahn-Strasse 15, 44227 Dortmund, Germany.
4
Bioanalytical Mass Spectrometry, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149 Münster, Germany.
5
Technische Universität Dresden, DFG-Research Center for Regenerative Therapies Dresden (CRTD), Fetscherstrasse 105, 01307 Dresden, Germany.
6
Technische Universität Dresden, DFG-Research Center for Regenerative Therapies Dresden (CRTD), Fetscherstrasse 105, 01307 Dresden, Germany; German Center for Neurodegenerative Diseases (DZNE) within the Helmholtz Association, Tatzberg 41, 01307 Dresden, Germany.
7
Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany.
8
Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149 Münster, Germany.
9
Department of Pediatrics, Division of Child Neurology, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, PA 15224, USA; Department of Human Genetics, University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA 15261, USA.
10
Department of Pediatrics, Division of Child Neurology, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, PA 15224, USA; Department of Human Genetics, University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA 15261, USA; Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
11
Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149 Münster, Germany; University of Münster, Medical Faculty, Domagkstrasse 3, 48149 Münster, Germany.
12
Technische Universität Dresden, DFG-Research Center for Regenerative Therapies Dresden (CRTD), Fetscherstrasse 105, 01307 Dresden, Germany; Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149 Münster, Germany. Electronic address: jared.sterneckert@tu-dresden.de.

Abstract

Neuroinflammation is a hallmark of neurological disorders and is accompanied by the production of neurotoxic agents such as nitric oxide. We used stem cell-based phenotypic screening and identified small molecules that directly protected neurons from neuroinflammation-induced degeneration. We demonstrate that inhibition of CDK5 is involved in, but not sufficient for, neuroprotection. Instead, additional inhibition of GSK3β is required to enhance the neuroprotective effects of CDK5 inhibition, which was confirmed using short hairpin RNA-mediated knockdown of CDK5 and GSK3β. Quantitative phosphoproteomics and high-content imaging demonstrate that neurite degeneration is mediated by aberrant phosphorylation of multiple microtubule-associated proteins. Finally, we show that our hit compound protects neurons in vivo in zebrafish models of motor neuron degeneration and Alzheimer's disease. Thus, we demonstrate an overlap of CDK5 and GSK3β in mediating the regulation of the neuronal cytoskeleton and that our hit compound LDC8 represents a promising starting point for neuroprotective drugs.

KEYWORDS:

ALS; CDK5; GSK3β; induced pluripotent stem cells; neurodegeneration; neuroinflammation; stem cell-based phenotypic screening

PMID:
30773488
DOI:
10.1016/j.stemcr.2019.01.015
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