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.

该项目的主要目的是阐明调节的功能机制神经系统通过蛋白O-甘露糖基化（POM）。 POM是具有O-糖基化的必不可少的类型从果蝇到多种动物的发展和生理的深远影响人类。尽管受POM影响的生物学功能的光谱很广，但到目前为止唯一的研究 POM的靶标是dystroglycan（DG）。 DG的POM修饰缺陷导致严重的肌肉营养不良称为营养不良的肿瘤病。与POM缺陷相关的病理机制很复杂，并且保持不当理解，尤其是在神经系统中。最近的研究表明POM 修饰会影响许多蛋白质的功能，这有助于多糖菌病。但是，除DG以外，POM在蛋白质上的功能在很大程度上未知。这糖基化的复杂性和体内方法的局限性为研究带来了重大挑战哺乳动物生物中的POM。在这里，我们提出了一个使用优势的多学科项目果蝇模型，包括强大的遗传方法的武器，简化糖基化和 POM和DG突变体的实验性不适，以阐明DG-的分子和细胞机制 POM的依赖和DG无关功能，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（RPTPS）是功能上重要的POM靶标，并揭示了POM调节通过影响感觉神经元与中枢神经系统之间的沟通来协调肌肉收缩。我们将利用这些结果，同时着重于三个具体目的：（1）分析pom在调节感觉神经元和协调的肌肉收缩。使用实时成像技术结合遗传和神经生物学方法，我们将全面研究感觉神经元，中枢神经系统细胞和肌肉之间的通信。（2）调查 RPTP功能的POM。使用体内和体外方法，我们将研究POM如何影响分子，细胞和生物水平的功能RPTP。（3）揭示POM的新分子靶标并阐明其在神经系统中的功能。我们将使用糖蛋白质组学方法来识别具有POM修饰的蛋白质。我们将分析POM在体内新靶标上的功能，重点在神经系统中起作用的蛋白质。我们预计该项目将建立新的范式 POM介导的神经系统调节，并将阐明新进化保守的DG- POM功能的依赖和独立机制，这将阐明与POM异常有关的人类疾病。