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Molecular mechanisms of Munc18-1 linked infantile seizure disorders and rational rescue strategies

Munc18-1相关婴儿癫痫发作的分子机制及合理抢救策略

基本信息

批准号：
9360727
负责人：
Jacqueline Burre
金额：
$ 35.92万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-15 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9360727
关键词：
APBA1 gene APBA2 gene Affect Animals Area Basic Science Binding Biochemical Biological Biophysics Caenorhabditis elegans Cells Chemicals Complex Data Defect Development Disease Docking Dominant-Negative Mutation Epilepsy Fostering Functional disorder Genes Genus Mentha Goals Impairment In Vitro Knock-out Knowledge Link Measures Medical Missense Mutation Mission Modeling Molecular Molecular Chaperones Mus Mutation Neuronal Dysfunction Neurons Nonsense Mutation Paralysed Phenotype Phenylbutyrates Process Proteins Public Health Recombinant Proteins Recombinants Recruitment Activity Regulation Research Role SNAP receptor Sorbitol Splice-Site Mutation Structure Symptoms Synapses Synaptic Vesicles Syndrome System Techniques Testing Therapeutic Intervention Translational Research Trehalose United States National Institutes of Health Variant Yeasts base epileptic encephalopathies fly functional loss functional restoration in vivo infancy innovation multidisciplinary mutant nervous system disorder neuronal survival neurotransmitter release novel novel therapeutics prevent synaptic function syntaxin 1 syntaxin A thermostability trafficking

项目摘要

Mutations in Munc18-1 are associated with three infantile epileptic encephalopathies, but the mechanistic relationship between mutations and these diseases is unknown. The long term goal is to clarify mechanisms by which specific synaptic dysfunctions trigger neurological disorders. The objective in this application is to determine how mutations in Munc18-1 cause neuronal defects, and to identify rescue strategies to reverse such deficits. Munc18-1 (also called STXBP1, SEC1 in yeast, Rop in flies, and unc18 in worms) controls neurotransmitter release at the synapse via binding to multiple effector proteins. Over 30 heterozygous de novo mu- tations have been identified in the Munc18-1 gene that cause the infantile epileptic encephalopathies Ohtahara, West, and Dravet syndrome, but it is unknown how these mutations affect neurons and trigger three different diseases. The central hypothesis, based on strong preliminary data, is that mutations in Munc18-1 result in defects in its folding, stability and localization, and elicit same defects in wild-type Munc18-1. This loss of functional Munc18-1 subsequently impairs the function of its effectors and triggers synaptic dysfunction, which can be restored by stabilizing Munc18-1. The rationale for these studies is that understanding of how mutations in Munc18-1 trigger synaptic dysfunction in infantile epileptic encephalopathies will create opportunities for the development of novel therapies beyond the current, limited symptom-based therapy. Guided by strong preliminary data, this hypothesis will be tested in three specific aims: 1) Determine the impact of disease-relevant mutations in Munc18-1 on protein stability; 2) Determine how Munc18-1 mutants affect the stability of its effectors and synapse function; and 3) Identify rescue strategies to stabilize Munc18-1 and restore its function. Under the first aim, stability, folding, aggregation and intracellular targeting will be quantified for Munc18-1 wild- type and mutants, combining purified recombinant proteins and primary neurons with biochemical and cell biological techniques. Under the second aim, stability, interaction, and targeting of Munc18-1's effectors syntaxin- 1, Doc2, Mint1, Mint2, and rab3, as well as synapse integrity and function will be analyzed, using purified proteins, primary neurons, and in vivo mouse and worm models. Under the third aim, chemical and molecular chaperones will be employed to restore deficits in Munc18-1 and in synapse structure and function, using same paradigms as for aims 1 and 2. This research is significant, because it will clarify the molecular mechanisms underlying Munc18-1-linked epilepsies, and will have translational importance in the development of new rational treatments. This research is innovative, because it 1) tests the novel hypothesis that Munc18-1 mutations cause synaptic dysfunction via a dominant-negative mechanism, 2) uses a multidisciplinary and systematic approach that has not previously been used in this research area, 3) is technically innovative because of newly generated C. elegans strains, and 4) shifts focus from a symptom-centered perspective to an approach that focusses on understanding convergent underlying disease mechanisms that pivot on Munc18-1.

Munc18-1的突变与三种婴儿癫痫性脑病有关，但突变与这些疾病之间的机制关系尚不清楚。长期目标是通过