Fasudil attenuates aggregation of α-synuclein in models of Parkinson's disease.

Lars Tatenhorst,Mathias Bähr,Jan C Koch,Vivian Dambeck,Hagen Walle,Tiago F Outeiro,Stefan Becker,Lars Tönges,Luis Fonseca-Ornelas,Tomás Lopes Da Fonseca,Markus Zweckstetter,Paul Lingor,Katrin Eckermann

doi:10.1186/s40478-016-0310-y

Abstract

Parkinson’s disease (PD) is the most common neurodegenerative movement disorder, yet disease-modifying treatments do not currently exist. Rho-associated protein kinase (ROCK) was recently described as a novel neuroprotective target in PD. Since alpha-synuclein (α-Syn) aggregation is a major hallmark in the pathogenesis of PD, we aimed to evaluate the anti-aggregative potential of pharmacological ROCK inhibition using the isoquinoline derivative Fasudil, a small molecule inhibitor already approved for clinical use in humans. Fasudil treatment significantly reduced α-Syn aggregation in vitro in a H4 cell culture model as well as in a cell-free assay. Nuclear magnetic resonance spectroscopy analysis revealed a direct binding of Fasudil to tyrosine residues Y133 and Y136 in the C-terminal region of α-Syn. Importantly, this binding was shown to be biologically relevant using site-directed mutagenesis of these residues in the cell culture model. Furthermore, we evaluated the impact of long-term Fasudil treatment on α-Syn pathology in vivo in a transgenic mouse model overexpressing human α-Syn bearing the A53T mutation (α-SynA53T mice). Fasudil treatment improved motor and cognitive functions in α-SynA53T mice as determined by CatwalkTM gait analysis and novel object recognition (NOR), without apparent side effects. Finally, immunohistochemical analysis revealed a significant reduction of α-Syn pathology in the midbrain of α-SynA53T mice after Fasudil treatment. Our results demonstrate that Fasudil, next to its effects mediated by ROCK-inhibition, directly interacts with α-Syn and attenuates α-Syn pathology. This underscores the translational potential of Fasudil as a disease-modifying drug for the treatment of PD and other synucleinopathies.Electronic supplementary materialThe online version of this article (doi:10.1186/s40478-016-0310-y) contains supplementary material, which is available to authorized users.

Highlights

In the coming decades the number of patients affected by neurodegenerative disorders, including Parkinson’s disease (PD), will inevitably increase [12]
Since α-Syn aggregation is a major step in the pathogenesis of PD, we evaluated the anti-aggregative potential of pharmacological Rho-associated protein kinase (ROCK) inhibition using the isoquinoline derivative Fasudil, a small molecule inhibitor already approved for clinical use in humans in Japan [44]
A concentration of 20 μM Fasudil was used in the ongoing cell culture experiments

Summary

Introduction

In the coming decades the number of patients affected by neurodegenerative disorders, including Parkinson’s disease (PD), will inevitably increase [12]. Neurodegeneration in PD is accompanied by the appearance of Lewy bodies, intra-neuronal protein aggregates with α-synuclein (α-Syn) as major component [4]. In previous studies in cell culture and animal models of PD, Rho-associated protein kinase (ROCK) was identified as novel molecular neuroprotective target [27, 50, 58, 59, 61]. Pharmacological ROCK inhibition protected against MPTP toxicity, attenuating dopaminergic cell death and increasing regenerative sprouting in vivo [59]. Since α-Syn aggregation is a major step in the pathogenesis of PD, we evaluated the anti-aggregative potential of pharmacological ROCK inhibition using the isoquinoline derivative Fasudil, a small molecule inhibitor already approved for clinical use in humans in Japan [44]

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