Alternative Splicing Modulates the Activity of CaV3.1. an Ion Channel Gene Involved in Spinocerebellar Ataxia, Epilepsy, and Autism Spectrum Disorders

选择性剪接调节 CaV3.1 的活性。

• 批准号: 10797338
• 负责人: Matteo Ruggiu
• 金额: $ 9.93万
• 依托单位: ST. JOHN'S UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-12 至 2025-09-11
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10797338
• 关键词: Absence Epilepsy; Action Potentials; Affect; Alternative Splicing; Binding; Biological Process; Biology; Biomedical Research; Biophysics; Bipolar Disorder; Brain; C-terminal; CACNA1G gene; Calcium; Calcium Channel; Calmodulin; Cardiac; Cell membrane; Cells; Cellular biology; Central Nervous System; Clinical; Data; Defect; Disease; Epilepsy; Event; Exons; Functional disorder; Gene Expression; Genes; Goals; Hormonal; Human; Ion Channel; Ion Channel Gating; Kinetics; Knockout Mice; Learning; Location; Malignant hyperpyrexia due to anesthesia; Maps; Membrane; Membrane Potentials; Messenger RNA; Methodology; Migraine; Molecular; Molecular Biology; Muscle; Mutate; Mutation; Neurons; Pathology; Patients; Permeability; Physiological; Physiological Processes; Physiology; Play; Potassium Channel; Property; Proteins; RNA; RNA Splicing; RNA-Binding Proteins; Regulation; Role; Shapes; Signal Transduction Pathway; Spinocerebellar Ataxias; Structure; Testing; Thalamic structure; Timothy syndrome; Tissues; Variant; autism spectrum disorder; bioinformatics pipeline; biophysical properties; career; cell type; design; differential expression; experimental study; graduate student; nervous system disorder; neurotransmitter release; novel therapeutic intervention; sensor; success; transcriptome sequencing; undergraduate student; voltage

PROJECT SUMMARY/ABSTRACT The central nervous system comprises the tissues and cells with the highest rate of alternative splicing in the body, and RNA-binding proteins play a major functional role in neurons. To better understand the contribution of RNA splicing to nerve cell biology, and to help elucidate the function that splicing plays in neuron physiology and neurologic disorders it is necessary to characterize how the inclusion or skipping of specific exons modulates the physiological properties of molecules — such as ion channels — that are critical for neuronal function, and to characterize how these splicing events are regulated at the cellular and molecular level. Our long-term goal is to understand the molecular mechanisms regulating protein-RNA networks that control alternative splicing in the brain, and how they relate to the biology of neurons and to disorders of the nervous system. The objective of this proposal is to study how alternative splicing of CaV3.1, a voltage-gated Calcium channel that significantly contributes to the regulation of cell membrane excitability — particularly in muscle and neurons — and that is mutated in patients with spinocerebellar ataxia-42 (SCA42) is regulated in different neuron cell types, and how it may contributes to the modulation of channel activity. The central hypothesis of this proposal is that neuronal cell type-specific alternative splicing of CaV3.1 at the C-terminus shapes the physiological properties of this voltage-gated ion channel. In Aim 1 we will test the hypothesis that CaV3.1 alternatively spliced exons are differentially expressed in different neuronal cell types in the brain. To tackle this question, we have developed an RNAseq-based bioinformatics pipeline that will allow us to interrogate differential splicing between neuronal subclasses defined at different hierarchical levels. This methodology will not only provide a snapshot of the alternative splicing landscape of CaV3.1 in different neuronal subclasses in the brain, but it will also allow us to generate predictions on how these alternative splicing events are regulated. In Aim 2 we will test the hypothesis that alternative splicing at the C-terminus significantly contributes to the regulation of the physiological activity of this ion channel. Since several disease-associated mutations in CaV3.1 map to alternatively spliced exons, understanding how alternative splicing modulates channel activity is critical. Since patients with CaV3.1-associated pathologies display defects in Calcium current properties, understanding how alternative splicing may modulate the biological functions of CaV3.1 and how this modulation is regulated, may have broad and significant clinical implications in spinocerebellar ataxia, epilepsy, and autism spectrum disorders, and it may inform the design of novel therapeutic strategies. Moreover, this project will provide both undergraduate and graduate students with a unique opportunity to learn the fundamentals of molecular biology and biomedical research and help them in their pursue of a career in the biomedical field.

项目摘要/摘要 中枢神经系统包含组织和细胞，其替代剪接速率是 身体和RNA结合蛋白在神经元中起着主要的功能作用。 RNA剪接以神经细胞生物学的剪接，并有助于阐明在神经元生理中发挥作用的剪接的结合物 和神经系统疾病有必要表征特定外显子的包含或跳过 调节分子的生理特性，例如离子通道 - 这对于神经元至关重要 功能，并表征剪接事件在细胞和分子水平上如何定制的。 长期目标是了解控制控制蛋白-RNA网络的分子机制 大脑中的替代剪接，以及与神经元的生物学和神经的生物学关系 系统。 大大有助于调节细胞膜兴奋性的通道 - 尤其是在肌肉中 和神经元 - 在脊椎动下共济失调-42（SCA42）的患者中静音。 神经元细胞类型，以及如何促进通道活性的调制。 该蛋白质是Cav3.1在C末端形状的Cav3.1的神经元细胞细胞替代剪接。 该电压门控离子通道的生理学特性。 在AIM 1中，我们将检验以下假设：Cav3.1替代剪接外显子在 大脑中的不同神经元细胞类型。 生物信息学管道将使我们能够在定义的神经元子类之间询问差异剪接 在不同的层次级别。 大脑中不同神经元子类中Cav3.1的景观 关于替代剪接事件的预测是在AIM 2中进行的。 C末端的替代剪接显着有助于调节生理活性 该离子通道。 了解替代剪接如何调节通道活动至关重要。 由于患有CAV3.1相关病理的患者在钙电流特性中显示缺陷，因此 了解替代剪接如何调节Cav3.1的生物学功能以及如何调节它 调节是规范的，可能在脊椎动下共济失调，癫痫，癫痫病中具有广泛而显着的临床意义。 和自闭症谱系障碍，并且可以为新型治疗策略的设计提供信息 为地下和研究生提供独特的机会来学习 分子生物学和生物医学研究的基本原理，并帮助他们从事Teer Teo的职业 生物医学领域。