Fasudil attenuates aggregation of α-synuclein in models of Parkinson's disease.
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-Syn(A53T) mice). Fasudil treatment improved motor and cognitive functions in α-Syn(A53T) mice as determined by Catwalk(TM) 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-Syn(A53T) 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.
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.
Lars
L
Tatenhorst
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.
Katrin
K
Eckermann
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.
Vivian
V
Dambeck
Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077, Göttingen, Germany.
Luis
L
Fonseca-Ornelas
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.
Hagen
H
Walle
Deparment of NeuroDegeneration and Restorative Research, University Medicine Göttingen, Waldweg 33, 37073, Göttingen, Germany.|Instituto de Fisiologia, Faculty of Medicine, University of Lisbon, Lisboa, 1649-028, Portugal.
Tomás
T
Lopes da Fonseca
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.|DFG Research Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Humboldtalle 23, 37073, Göttingen, Germany.
Jan C
JC
Koch
Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077, Göttingen, Germany.
Stefan
S
Becker
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.|Current address: Neurological Clinic, St. Josef-Hospital, Ruhr-University Bochum, Gudrunstr. 56, 44791, Bochum, Germany.
Lars
L
Tönges
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany.|DFG Research Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Humboldtalle 23, 37073, Göttingen, Germany.
Mathias
M
Bähr
Deparment of NeuroDegeneration and Restorative Research, University Medicine Göttingen, Waldweg 33, 37073, Göttingen, Germany.|DFG Research Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Humboldtalle 23, 37073, Göttingen, Germany.
Tiago F
TF
Outeiro
Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077, Göttingen, Germany.|DFG Research Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Humboldtalle 23, 37073, Göttingen, Germany.|German Center for Neurodegenerative Diseases (DZNE), Am Fassberg 11, 37077, Göttingen, Germany.
Markus
M
Zweckstetter
Deparment of Neurology, University Medicine Göttingen, Robert-Koch-Str. 40, 37075, Göttingen, Germany. plingor@gwdg.de.|DFG Research Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Humboldtalle 23, 37073, Göttingen, Germany. plingor@gwdg.de.
Paul
P
Lingor
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D000577:Amides / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D001921:Brain / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D002352:Carrier Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010300:Parkinson Disease / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D066329:Protein Aggregates / Q000187:drug effects* / Q000235:genetics*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D011597:Psychomotor Performance / Q000187:drug effects; D011725:Pyridines / Q000494:pharmacology / Q000627:therapeutic use; D021641:Recognition, Psychology / Q000187:drug effects; D013997:Time Factors; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
D000577:Amides; D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D009419:Nerve Tissue Proteins; D066329:Protein Aggregates; D047428:Protein Kinase Inhibitors; D011725:Pyridines; C118958:SNCAIP protein, human; D051844:alpha-Synuclein; C108830:Y 27632; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D014446:Tyrosine 3-Monooxygenase; C049347:fasudil
A53T mouse model; Fasudil; Parkinson’s disease; α-synuclein aggregation