Functional mechanisms underlying Dystroglycan-dependent and independent roles of protein O-mannosylation in the nervous system

蛋白质 O-甘露糖基化在神经系统中依赖和独立作用的功能机制

基本信息

批准号：
9384393
负责人：
VLADISLAV M PANIN
金额：
$ 31.31万
依托单位：
TEXAS A&M AGRILIFE RESEARCH
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：
2017-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9384393
关键词：
Affect Afferent Neurons Animals Area Axon Biochemical Bioinformatics Biological Biological Process Biology Biomedical Research Cells Clinical Research Communication Complex Data Defect Development Developmental Biology Diagnostic Disease Drosophila genus Dystroglycan Embryo Genes Genetic Glycobiology Glycoproteins Hereditary Disease Human Imaging Techniques In Vitro Investigation Light Mediating Mediator of activation protein Membrane Proteins Methods Modeling Modernization Modification Molecular Molecular Target Muscle Muscle Contraction Muscular Dystrophies Nervous system structure Neurobiology Organism Outcome Pathogenicity Pathway interactions Pattern Phenotype Physiology Play Polysaccharides Post-Translational Protein Processing Posture Protein Tyrosine Phosphatase Proteins Regulation Research Role Severities Structure Testing Therapeutic base dystroglycanopathy genetic approach glycoproteomics glycosylation human disease in vivo innovation multidisciplinary mutant neuromuscular novel novel therapeutics protein function receptor receptor function

项目摘要

The main objective of this project is to elucidate functional mechanisms underlying regulation of the nervous system by protein O-mannosylation (POM). POM is an essential type of O-glycosylation that has a profound effect on the development and physiology in a broad range of animals, from Drosophila to humans. Although the spectrum of biological functions affected by POM is wide, so far the only well-studied target of POM is Dystroglycan (Dg). Defects in POM modifications of Dg result in severe muscular dystrophies called dystroglycanopathies. Pathomechanisms associated with POM defects are complex and remain poorly understood, particularly in the nervous system. Recent studies suggested that POM modification affects functions of many proteins, which contributes to pathogenic mechanisms of dystroglycanopathies. However, functions of POM on proteins besides Dg are largely unknown. The complexity of glycosylation and limitations of in vivo approaches create significant challenges for studying POM in mammalian organisms. Here we propose a multidisciplinary project that uses advantages of Drosophila model, including powerful arsenal of genetic approaches, simplified glycosylation and experimental amenability of POM and Dg mutants, to elucidate molecular and cellular mechanisms of Dg- dependent and Dg-independent functions of POM, with the focus on the nervous system and neuromuscular development and physiology. Our preliminary studies suggested that Receptor Protein Tyrosine Phosphatases (RPTPs) are functionally important POM targets and revealed that POM regulates coordinated muscle contractions by affecting communication between sensory neurons and the CNS. We will capitalize on these results while focusing on three specific aims: (1) To analyze the role of POM in regulation of sensory neurons and coordinated muscle contractions. Using live imaging techniques combined with genetic and neurobiological approaches, we will comprehensively investigate the role of POM in communication between sensory neurons, CNS cells and muscles. (2) To investigate the effect of POM on RPTP function. Using in vivo and in vitro approaches, we will investigate how POM affects functions RPTPs at molecular, cellular, and organismal levels. (3) To reveal new molecular targets of POM and elucidate their function in the nervous system. We will use glycoproteomic approaches to identify proteins with POM modifications. We will analyze functions of POM on novel targets in vivo, focusing on proteins that function in the nervous system. We anticipate that this project will establish new paradigms of POM-mediated regulation of the nervous system and will elucidate new evolutionarily conserved, Dg- dependent and independent mechanisms of POM functions, which will shed light on pathomechanisms of human diseases associated with POM abnormalities.

该项目的主要目标是阐明监管的功能机制蛋白质 O-甘露糖基化 (POM) 的神经系统。 POM 是一种重要的 O-糖基化类型，具有对从果蝇到多种动物的发育和生理机能产生深远影响人类。尽管 POM 影响的生物功能范围很广，但迄今为止唯一得到充分研究的 POM 的目标是肌营养不良聚糖 (Dg)。 Dg 的 POM 修饰缺陷会导致严重的肌肉损伤营养不良称为肌营养不良病。与 POM 缺陷相关的病理机制非常复杂人们对它的了解仍然知之甚少，特别是在神经系统中。最近的研究表明 POM 修饰会影响许多蛋白质的功能，从而导致致病机制肌营养不良症。然而，POM 对除 Dg 之外的蛋白质的功能尚不清楚。这糖基化的复杂性和体内方法的局限性给研究带来了重大挑战哺乳动物体内的 POM。在这里，我们提出了一个利用以下优点的多学科项目：果蝇模型，包括强大的遗传方法库、简化的糖基化和 POM 和 Dg 突变体的实验适应性，以阐明 Dg- 的分子和细胞机制 POM 的依赖和 Dg 独立功能，重点是神经系统和神经肌肉发育和生理学。我们的初步研究表明受体蛋白酪氨酸磷酸酶 (RPTP) 是功能上重要的 POM 靶标，并揭示 POM 调节通过影响感觉神经元和中枢神经系统之间的通讯来协调肌肉收缩。我们将利用这些结果，同时重点关注三个具体目标：（1）分析 POM 在感觉神经元的调节和协调的肌肉收缩。使用实时成像技术结合遗传学和神经生物学方法，我们将全面研究 POM 在感觉神经元、中枢神经系统细胞和肌肉之间的通讯。 (2) 考察效果 POM 上的 RPTP 功能。使用体内和体外方法，我们将研究 POM 如何影响 RPTP 在分子、细胞和有机体水平上发挥作用。 (3)揭示POM新分子靶点并阐明它们在神经系统中的功能。我们将使用糖蛋白组学方法来鉴定具有 POM 修饰的蛋白质。我们将分析 POM 对体内新靶标的功能，重点关注在神经系统中发挥作用的蛋白质。我们预计该项目将建立新的范式 POM 介导的神经系统调节，并将阐明新的进化保守的 Dg- POM 功能的依赖和独立机制，这将有助于阐明 POM 功能的病理机制与 POM 异常相关的人类疾病。