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

• DOI: 10.1016/j.stemcr.2019.01.015
• Publication Date: 2019-02-14T15:42:28Z
• Authors: Lydia Reinhardt; Susanne Kordes; Peter Reinhardt; Michael Glatza; Matthias Baumann; Hannes C.A. Drexler; Sascha Menninger; Gunther Zischinsky; Jan Eickhoff; Claudia Fröb; Prabesh Bhattarai; Guruchandar Arulm...; Lara Marrone; Antje Janosch; Kenjiro Adachi; Martin Stehling; Eric N. Anderson; Masin Abo-Rady; Marc Bickle; Udai Bhan Pandey; Michell M. Reimer; Caghan Kizil; Hans R. Schöler; Peter Nussbaumer; Bert Klebl; Jared L. Sterneckert
• Keywords: drug effects [Signal Transduction]; Medicine (General); metabolism [Cytoskeleton]; QH301-705.5; metabolism [Glycogen Synthase Kinase 3 beta]; metabolism [Microtubules]; drug effects [Neurites]; Microtubules; Article; 03 medical and health sciences; R5-920; drug effects [Phosphorylation]; Alzheimer Disease; Neurites; drug therapy [Alzheimer Disease]; drug effects [Neurons]; Animals; Humans; ddc:610; metabolism [Zebrafish]; Biology (General); Phosphorylation; metabolism [Nerve Degeneration]; Cytoskeleton; Zebrafish; Inflammation; Neurons; metabolism [Inflammation]; 0303 health sciences; Glycogen Synthase Kinase 3 beta; pharmacology [Neuroprotective Agents]; drug therapy [Inflammation]; metabolism [Neurites]; Cyclin-Dependent Kinase 5; 3. Good health; drug effects [Cytoskeleton]; Neuroprotective Agents; drug effects [Microtubules]; metabolism [Neurons]; Nerve Degeneration; metabolism [Cyclin-Dependent Kinase 5]; drug therapy [Nerve Degeneration]; metabolism [Alzheimer Disease]; Signal Transduction

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.