RNA Binding Proteins in Complex Neurological Disease

复杂神经系统疾病中的 RNA 结合蛋白

基本信息

批准号：
8858948
负责人：
WAYNE N. FRANKEL
金额：
$ 39.14万
依托单位：
JACKSON LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-02-01 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8858948
关键词：
Accounting Action Potentials Address Adult Animal Model Animals Autistic Disorder Back Behavior Binding Binding Sites Brain Brain Diseases Cell Line Cells Central Nervous System Diseases Cerebral cortex Clustered Regularly Interspaced Short Palindromic Repeats Complex Cues Cytoplasmic Granules Data Development Disease Dissection Dose Drosophila genus Employee Strikes Epilepsy Equilibrium Etiology FMR1 FMRP Failure Family Fragile X Mental Retardation Protein Fragile X Syndrome Functional disorder Funding Gene Expression Profile Gene Targeting Genes Genetic Genetic Translation Genomic approach Genotype Hippocampus (Brain)Homeostasis Human Hyperactive behavior Individual Intellectual functioning disability Ion Channel Lead Learning Location Membrane Potentials Mental disorders Messenger RNA Metabolism Mild obesity Modeling Molecular Mus Mutagenesis Mutant Strains Mice Mutate Nature Neurologic Neurons Neuropil Neurotransmitter Receptor Orthologous Gene Outcome Pathology Patients Pattern Phenotype Physiological Polyribosomes Predisposition Proteins RNA RNA Binding RNA-Binding Proteins Recycling Regulation Research Role SCN8A gene Schizophrenia Seizures Signal Transduction Social Interaction Sodium Channel Sucrose Symptoms Synapses Synaptic Vesicles Synaptic plasticity System Testing Transgenes Translating Translations Untranslated Regions Variant Vision Work complex biological systems density disability excitatory neuron genetic variant genome sequencing in vivo interest member mutant nervous system disorder neuron development neuronal cell body neuronal excitability neurotransmission non-genetic particle postnatal programs public health relevance social success synaptic function transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Many factors make genetically complex diseases complex. Classically they are defined as an interaction between multiple genetic variants and non-genetic factors. Progress in genome sequencing and within-species variation has generated much interest in identifying polygenic variants from human and model organisms, with some success. But one cannot lose sight of the importance of physiological complexity; even Mendelian variants can wreak havoc when operating in a complex biological system. For functional phenotypes, such as excitability disorders of the CNS, this concept is understudied. Epilepsy is genetically complex to be sure, but as the canonical excitability disorder of the brain it also serves as a leading example for approaching other, harder-to-crack functional disorders, such as autism and schizophrenia, that are also likely to have excito-pathology at their cores. Neuronal excitability is determined primarily by molecules, such as ion channels and transporters, neurotransmitter receptors, and synaptic proteins, controlling membrane potential and synaptic signaling in order to achieve an appropriate balance of excitation and inhibition. Although cis-variants in genes encoding these molecules can lead to specific phenotypes, trans-factors that regulate their expression must be critical for maintaining this balance at a higher, coordinated level. We previously identified and characterized hypomorphic and null genotypes in Celf4 (formerly known as Brunol4), encoding a brain-specific member of the BRUNO/CUGBP/CELF family of RNA binding proteins. Celf4 mutants have a complex seizure disorder and other neurological phenotypes, such as hyperactivity, mild obesity and abnormal social interaction. Very recently human CELF4 deficiency revealed these and additional symptoms, such as intellectual disability. In our initial funding period, we found that CELF4 is most tightly associated with very high-density RNA granule particles and targets a vast number of mRNAs in excitatory neurons. Many targets are involved in synaptic functions, and they tend to be dysregulated within neurons of mutant mice - in all directions, but with a tendency towards increased expression away from the cell body. These findings are consistent with a role for CELF4 in control "translational silencing" perhaps at local, subcellular levels. We also obtained evidence for CELF4 effects on intrinsic neuronal hyperexcitation, via increased expression of sodium channel Nav1.6, and system-wide dysregulation via impaired homeostatic plasticity presumably due to dysregulation of synaptic proteins. Our hypothesis is that the combination of such effects accounts for full-blown disease. In the next 5 years, our research will address two prevailing themes that work together to address this idea. The first addresses the pattern of CELF4 binding motifs within the 3'-UTR of target mRNAs, and the consequences of altering the motifs on mRNA abundance, localization and translation, both transcriptome-wide and for selected targets. The second theme uses in vivo mutagenesis to assess the contribution of individual targets to the multigenic etiology of complex neurological phenotypes.

描述（由申请人提供）：许多因素使遗传复杂疾病变得复杂，它们被定义为多种遗传变异和非遗传因素之间的相互作用。基因组测序和种内变异的进展引起了人们对识别多基因变异的兴趣。但人们不能忽视生理复杂性的重要性；即使是孟德尔变异在复杂的生物系统中运行时也会造成严重破坏，例如中枢神经系统的兴奋性障碍。癫痫病的遗传机制确实很复杂，但作为一种典型的大脑兴奋性障碍，它也可以作为解决其他更难破解的功能障碍（例如自闭症和精神分裂症）的主要例子。其核心具有兴奋病理学，神经元兴奋性主要由控制膜电位和突触的分子决定，例如离子通道和转运蛋白、神经递质受体和突触蛋白。尽管编码这些分子的基因中的顺式变体可以导致特定的表型，但调节其表达的反式因子对于将这种平衡维持在更高的协调水平至关重要。先前在 Celf4（以前称为 Brunol4）中鉴定并表征了低效基因型和无效基因型，编码了 RNA 结合蛋白 BRUNO/CUGBP/CELF 家族的大脑特异性成员 Celf4 突变体具有复杂的癫痫症和其他神经系统疾病。表型，例如多动、轻度肥胖和异常社交互动，最近人类 CELF4 缺陷揭示了这些和其他症状，例如智力障碍。在我们最初的资助期间，我们发现 CELF4 与非常高密度的 RNA 颗粒密切相关。颗粒并靶向兴奋性神经元中的大量 mRNA，许多靶点参与突触功能，并且它们在突变小鼠的神经元内往往在各个方向上失调，但有增加远离细胞体的表达的趋势。研究结果与 CELF4 可能在局部、亚细胞水平上控制“翻译沉默”的作用一致。我们还获得了 CELF4 通过钠通道 Nav1.6 表达增加和系统范围失调对内在神经元过度兴奋的影响的证据。稳态可塑性可能是由于突触蛋白的失调造成的，我们的假设是，这些效应的结合导致了全面的疾病。在接下来的五年中，我们的研究将解决两个共同的主题。第一个主题涉及目标 mRNA 3'-UTR 内的 CELF4 结合基序的模式，以及改变基序对转录组范围内和选定目标的 mRNA 丰度、定位和翻译的影响。使用体内诱变来评估单个靶标对复杂神经表型的多基因病因学的贡献。