Reduction of Phosphorylated Tau in Alzheimer’s Disease Induced Pluripotent Stem Cell-Derived Neuro-Spheroids by Rho-Associated Coiled-Coil Kinase Inhibitor Fasudil

Article type: Research Article

Authors: Giunti, Elisa^{a; b} | Collu, Roberto^{a; b} | Daley, Sarah^{a; b} | Querfurth, Henry^c | Morin, Peter^d | Killick, Richard^e | Melamed, Rachel D.^f | Xia, Weiming^{a; b; f; *}

Affiliations: [a] Geriatric Research Education and Clinical Center, Bedford VA Healthcare System, Bedford, MA, USA | [b] Department of Pharmacology, Physiology & Biophysics, Boston University Chobanian & Avedisian School of Medicine, Boston, MA, USA | [c] Department of Neurology, Tufts Medical Center, Boston, MA, USA | [d] Department of Neurology, Boston University Chobanian & Avedisian School of Medicine, Boston, Boston, MA, USA | [e] Maurice Wohl Clinical Neuroscience Institute, King’s College London, London, UK | [f] Department of Biological Sciences, Kennedy College of Sciences, University of Massachusetts, Lowell, MA, USA

Correspondence: [*] Correspondence to: Weiming Xia, PhD, 200 Springs Road, Building 70-202, Bedford VA Healthcare System, Bedford, MA 01730, USA. E-mail: [email protected].

Abstract: Background:Alzheimer’s disease (AD) is the most predominant form of dementia. Rho-associated coiled coil kinase (ROCK) inhibitor, fasudil, is one of the candidate drugs against the AD progression. Objective:We aimed to investigate possible changes of AD associated markers in three-dimensional neuro-spheroids (3D neuro-spheroids) generated from induced pluripotent stem cells derived from AD patients or healthy control subjects (HC) and to determine the impact of pharmacological intervention with the ROCK inhibitor fasudil. Methods:We treated 3D neuro-spheroids with fasudil and tested the possible effect on AD markers by ELISA, transcriptomic and proteomic analyses. Results:Transcriptomic analysis revealed a reduction in the expression of AKT serine/threonine-protein kinase 1 (AKT1) in AD neuro-spheroids, compared to HC. This decrease was reverted in the presence of fasudil. Proteomic analysis showed up- and down-regulation of proteins related to AKT pathway in fasudil-treated neuro-spheroids. We found an evident increase of phosphorylated tau at four different residues (pTau181, 202, 231, and 396) in AD compared to HC-derived neuro-spheroids. This was accompanied by a decrease of secreted clusterin (clu) and an increase of intracellular clu levels in AD patient-derived neuro-spheroids. Increases of phosphorylated tau in AD patient-derived neuro-spheroids were suppressed in the presence of fasudil. Conclusions:Fasudil modulates clu protein levels and enhances AKT1 that results in the suppression of AD associated tau phosphorylation.

Keywords: Alzheimer’s disease, fasudil, induced pluripotent stem cells, ROCK

DOI: 10.3233/JAD-230551

Journal: Journal of Alzheimer's Disease, vol. 96, no. 4, pp. 1695-1709, 2023