O-glycosylation mechanisms of neurological deficits in congenital disorders of glycosylation

先天性糖基化障碍神经功能缺损的O-糖基化机制

基本信息

批准号：
10250486
负责人：
Andrew Charles Edmondson
金额：
$ 18.08万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10250486
关键词：
Affect Astrocytes Biology Brain Career Mobility Cell Line Cells Cellular Morphology Complex Congenital disorders of glycosylation Constitutional Development Development Plans Developmental Delay Disorders Disease Epilepsy Event Exhibits Family member Foundations Functional disorder Funding Future Genetic Genetic Diseases Glycobiology Glycoproteins Goals Human Individual International Investigation Knockout Mice LoxP-flanked allele Mass Spectrum Analysis Mediating Mentors Microcephaly Modeling Molecular Morphology Mucins Mus Mutation Nervous System Physiology Nervous system structure Neurobiology Neurologic Neurologic Deficit Neurologic Dysfunctions Neurons Neurosciences Pathway interactions Patients Phenotype Physicians Post-Translational Protein Processing Process Protein Isoforms Proteins Rare Diseases Research Role Scientist Signal Transduction Site Symptoms Techniques Therapeutic Training behavioral phenotyping career development cell motility cell type comparative conditional knockout design glycoproteomics glycosylation glycosyltransferase in vitro Assay insight knowledge base molecular phenotype motor deficit mouse model nervous system disorder neurotransmitter release programs protein function rare genetic disorder relating to nervous system sensory integration skills tool

项目摘要

PROJECT SUMMARY/ABSTRACT Glycosylation is an essential, post-translational modification with complex and poorly understood roles in protein function. My long-term objective is to elucidate the neurobiological functions of glycosylation, including identifying the roles of critical glycosylation sites in neuronal protein function. The importance of glycosylation is emphasized by the congenital disorders of glycosylation (CDG), a group of genetic disorders that disrupt cellular glycosylation machinery. Affected patients exhibit severe neurological deficits. The genetic basis of CDG provides an opportunity to identify the neurobiological functions of glycosylation using mouse models and glycoproteomics. Understanding glycosylation in the nervous system will elucidate the pathophysiology of CDGs and other neurological diseases, enable therapeutic advances targeting glycosylation pathways, and inform normal function of glycosylation. GALNT2-CDG is a new CDG type caused by biallelic mutations in GALNT2, which encodes a critical glycosyltransferase initiating the first step in mucin-type O-glycosylation. GALNT2-CDG patients suffer from epilepsy and global developmental delay. Galnt2 constitutional knock-out mice recapitulate many of the patient neurological deficits. My central hypothesis is that site-specific loss of O-glycosylation on neural proteins contributes to neurological dysfunction. The specific objective of this project is to identify the cause of neurological dysfunction in GALNT2-CDG. This will be achieved by determining cellular origins of Galnt2 deficiency-mediated neurological deficits using Cre-mediated deletion of Galnt2 in neural cells and by identifying disrupted O-glycosylation in these cells using glycoproteomics. This proposed five-year career development plan focuses on achieving four objectives: develop research skills in mouse models and glycoproteomics, increase my knowledgebase in neuroscience and glycobiology, establish a body of work focusing on the role of glycosylation in the context of neurobiology, and obtain the necessary skills to transition to independence. Mentoring will be provided by Dr. Zhaolan Zhou, a recognized leader in the development and investigation of mouse models of genetic disorders that affect brain development and function, and Dr. Benjamin Garcia, a recognized expert in developing quantitative mass spectrometry techniques to interrogate post-translational modifications. The skill set developed through these investigations and career development plan will make me uniquely poised to uncover glycosylation-mediated mechanisms of CDG and other neurological diseases, as well as to elucidate the critical roles of glycosylation in human neurological function. These studies will generate new tools and a foundation to establish a long-term research program to investigate the pathophysiology of various glycosylation-related disorders in the nervous system and prepare me to become an independent R01-funded physician scientist.

项目摘要/摘要糖基化是一种必不可少的翻译后修饰，在复杂且了解不足的作用蛋白质功能。我的长期目标是阐明糖基化的神经生物学功能，包括鉴定临界糖基化位点在神经元蛋白功能中的作用。糖基化的重要性糖基化的先天性疾病（CDG）强调了，这是一组破坏的遗传疾病细胞糖基化机制。受影响的患者表现出严重的神经系统缺陷。遗传基础 CDG提供了使用小鼠模型和糖蛋白质组学。了解神经系统中的糖基化将阐明 CDG和其他神经系统疾病，使靶向糖基化途径的治疗进展以及告知糖基化的正常功能。 GALNT2-CDG是由Galnt2中的双重突变引起的一种新的CDG类型，它编码关键糖基转移酶启动了粘蛋白型O-糖基化的第一步。 GALNT2-CDG患者患有癫痫和全球发育延迟。 GALNT2宪法敲除小鼠概括了许多患者神经缺陷。我的中心假设是神经蛋白上的位点特异性O-糖基化丧失导致神经功能障碍。该项目的具体目的是确定 GALNT2-CDG中的神经功能障碍。这将通过确定galnt2的细胞起源来实现使用CRE介导的GALNT2在神经细胞中和通过使用糖蛋白酶学鉴定这些细胞中的O-糖基化破坏。该拟议的五年职业发展计划重点是实现四个目标：发展研究技能在小鼠模型和糖蛋白质组学中，增加了我在神经科学和糖脂学方面的知识基础，建立一个专注于糖基化在神经生物学背景下的作用的工作，并获得过渡到独立的必要技能。指导将由公认的Zhaolan Zhou博士提供影响大脑的遗传疾病小鼠模型的发展和调查的领导者开发和功能，以及开发定量质量的公认专家本杰明·加西亚（Benjamin Garcia）博士光谱技术来询问翻译后修饰。通过这些调查和职业发展计划开发的技能将使我独一无二准备发现CDG和其他神经系统疾病的糖基化介导的机制，以及阐明糖基化在人神经功能功能中的关键作用。这些研究将产生新的工具和建立长期研究计划的基础，以研究各种的病理生理神经系统中与糖基化相关的疾病，并准备成为独立的R01资助医师科学家。