TBC1D24-TLDc-related epilepsy exercise-induced dystonia: Rescue by antioxidants in a disease model
Journal
Brain
Journal Volume
142
Journal Issue
8
Pages
2319-2335
Date Issued
2019
Author(s)
L?thy K.
Mei D.
Fischer B.
De Fusco M.
Swerts J.
Paesmans J.
Parrini E.
Lubarr N.
Meijer I.A.
Mackenzie K.M.
Cittaro D.
Aridon P.
Schoovaerts N.
Vers?es W.
Verstreken P.
Casari G.
Guerrini R.
Abstract
Genetic mutations in TBC1D24 have been associated with multiple phenotypes, with epilepsy being the main clinical manifestation. The TBC1D24 protein consists of the unique association of a Tre2/Bub2/Cdc16 (TBC) domain and a TBC/lysin motif domain/catalytic (TLDc) domain. More than 50 missense and loss-of-function mutations have been described and are spread over the entire protein. Through whole genome/exome sequencing we identified compound heterozygous mutations, R360H and G501R, within the TLDc domain, in an index family with a Rolandic epilepsy exercise-induced dystonia phenotype (http://omim.org/entry/608105). A 20-year long clinical follow-up revealed that epilepsy was self-limited in all three affected patients, but exercise-induced dystonia persisted into adulthood in two. Furthermore, we identified three additional sporadic paediatric patients with a remarkably similar phenotype, two of whom had compound heterozygous mutations consisting of an in-frame deletion I81-K84 and an A500V mutation, and the third carried T182M and G511R missense mutations, overall revealing that all six patients harbour a missense mutation in the subdomain of TLDc between residues 500 and 511. We solved the crystal structure of the conserved Drosophila TLDc domain. This allowed us to predict destabilizing effects of the G501R and G511R mutations and, to a lesser degree, of R360H and potentially A500V. Next, we characterized the functional consequences of a strong and a weak TLDc mutation (TBC1D24G501R and TBC1D24R360H) using Drosophila, where TBC1D24/Skywalker regulates synaptic vesicle trafficking. In a Drosophila model neuronally expressing human TBC1D24, we demonstrated that the TBC1D24G501R TLDc mutation causes activity-induced locomotion and synaptic vesicle trafficking defects, while TBC1D24R360H is benign. The neuronal phenotypes of the TBC1D24G501R mutation are consistent with exacerbated oxidative stress sensitivity, which is rescued by treating TBC1D24G501R mutant animals with antioxidants N-acetylcysteine amide or α-tocopherol as indicated by restored synaptic vesicle trafficking levels and sustained behavioural activity. Our data thus show that mutations in the TLDc domain of TBC1D24 cause Rolandic-type focal motor epilepsy and exercise-induced dystonia. The humanized TBC1D24G501R fly model exhibits sustained activity and vesicle transport defects. We propose that the TBC1D24/Sky TLDc domain is a reactive oxygen species sensor mediating synaptic vesicle trafficking rates that, when dysfunctional, causes a movement disorder in patients and flies. The TLDc and TBC domain mutations' response to antioxidant treatment we observed in the animal model suggests a potential for combining antioxidant-based therapeutic approaches to TBC1D24-associated disorders with previously described lipid-altering strategies for TBC domain mutations. ? 2019 The Author(s) (2019). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved. For Permissions, please email: journals.permissions@oup.com.
Subjects
exercise-induced dystonia; oxidative stress; Rolandic epilepsy; TBC1D24
SDGs
Other Subjects
acetazolamide; acetylcysteine derivative; alpha tocopherol; brain protein; carbamazepine; carbidopa plus levodopa; clobazam; clonazepam; flunarizine; lacosamide; lamotrigine; n acetylcysteine amide; oxcarbazepine; reactive oxygen metabolite; tbc1d24 protein; trihexyphenidyl; ubidecarenone; unclassified drug; acetylcysteine; alpha tocopherol; antioxidant; Drosophila protein; guanosine triphosphatase activating protein; N-Acetylcysteinamide; Rab protein; reactive oxygen metabolite; recombinant protein; sky protein, Drosophila; TBC1D24 protein, human; adolescent; adult; animal experiment; animal model; anticonvulsant therapy; Article; behavior; cell migration; child; clinical article; controlled study; crystal structure; disease model; dystonia; electroencephalography; exercise; female; follow up; fruit fly model; gene mutation; hand tremor; heterozygosity; human; locomotion; male; missense mutation; nonhuman; nystagmus; phenotype; priority journal; protein stability; remission; rolandic epilepsy; school child; seizure; synapse vesicle; transport vesicle; whole exome sequencing; young adult; amino acid sequence; animal; case report; chemistry; disease model; Drosophila melanogaster; drug effect; dystonia; enzyme active site; gene deletion; genetics; infant; metabolism; molecular model; nerve cell; oxidative stress; pedigree; physiology; preschool child; protein conformation; protein motif; rolandic epilepsy; sequence alignment; sequence homology; transgenic animal; transport at the cellular level; X ray crystallography; Acetylcysteine; Adolescent; alpha-Tocopherol; Amino Acid Motifs; Amino Acid Sequence; Animals; Animals, Genetically Modified; Antioxidants; Biological Transport; Catalytic Domain; Child; Child, Preschool; Crystallography, X-Ray; Disease Models, Animal; Drosophila melanogaster; Drosophila Proteins; Dystonia; Epilepsy, Rolandic; Female; GTPase-Activating Proteins; Humans; Infant; Locomotion; Male; Models, Molecular; Mutation, Missense; Neurons; Oxidative Stress; Pedigree; Physical Exertion; Protein Conformation; rab GTP-Binding Proteins; Reactive Oxygen Species; Recombinant Proteins; Sequence Alignment; Sequence Deletion; Sequence Homology, Amino Acid; Synaptic Vesicles
Publisher
Oxford University Press
Type
journal article